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Sample records for actinomycete streptomyces sp

  1. Antibacterial metabolites from the Actinomycete Streptomyces sp. P294.

    PubMed

    Su, Huining; Shao, Hongwei; Zhang, Keqin; Li, Guohong

    2016-02-01

    The Actinomycete strain P294 was isolated from soil and identified as Streptomyces sp. based upon the results of 16S rRNA sequence analysis. Three compounds obtained from the solid fermentation products of this strain have been determined by 1D, 2D NMR and HRMS experiments. These compounds include two new compounds angumycinones C (1) and D (2), and the known compound X-14881 E (3). All compounds were assayed for antibacterial and nematicidal activity. The results showed the three compounds had different degrees of inhibitory activity against several target bacteria but no significant toxicity against the nematode Caenorhabditis elegans.

  2. Streptomyces lopnurensis sp. nov., an actinomycete isolated from soil.

    PubMed

    Zheng, Bei; Han, Xiao-Xue; Xia, Zhan-Feng; Wan, Chuan-Xing; Zhang, Li-Li

    2014-12-01

    A novel actinomycete, designated strain TRM 49590(T), was isolated from a soil sample from Lop Nur in Xinjiang Province, China. Strain TRM 49590(T) was aerobic, Gram-staining-positive, with an optimum NaCl concentration for growth of 1.5 % (w/v) and an optimum temperature for growth of 28-37 °C. The aerial mycelium was sparse, cylindrical and smooth-surfaced with irregular branches on ISP medium 4. The whole-cell sugars of strain TRM 49590(T) were ribose and glucose. The diagnostic diamino acid contained ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6) and MK-9(H8), with MK-9(H4) and MK-10(H6) present in smaller amounts. The major fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 62.2 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain TRM 49590(T) belongs to the genus Streptomyces with a sequence similarity of 97.16 % with the most closely related species Streptomyces sodiiphilus. Based on these observations, strain TRM 49590(T) is proposed to represent a novel species of the genus Streptomyces for which the name Streptomyces lopnurensis sp. nov. is suggested. The type strain is TRM 49590(T) ( = CCTCC AA 2013018(T) = NRRL B59109(T)).

  3. Streptomyces koyangensis sp. nov., a novel actinomycete that produces 4-phenyl-3-butenoic acid.

    PubMed

    Lee, Jee Yeon; Lee, Jung Yeop; Jung, Ho Won; Hwang, Byung Kook

    2005-01-01

    A 4-phenyl-3-butenoic acid-producing actinomycete, designated strain VK-A60T, was isolated from a soil sample collected from Koyang, Korea. Morphological and chemical characteristics of the strain were consistent with those of the genus Streptomyces. The cell wall of the strain contains LL-diaminopimelic acid. The predominant fatty acids are anteiso-C(15 : 0), iso-C(16 : 0) and C(16 : 0). The strain formed a distinct monophyletic line within the 16S rRNA gene sequence phylogenetic tree. Analyses of its morphological, physiological and biochemical characteristics, together with random amplified polymorphic DNA and DNA-DNA relatedness data, confirmed that strain VK-A60T represents a novel Streptomyces taxon that is distinguishable from closely related reference strains. Strain VK-A60T (=KCCM 10555T=NBRC 100598T) is proposed as the type strain of a novel species, for which the name Streptomyces koyangensis sp. nov. is proposed.

  4. Streptomyces gamaensis sp. nov., a novel actinomycete with antifungal activity isolated from soil in Gama, Chad.

    PubMed

    Zhao, Shanshan; Ye, Lan; Liu, Chongxi; Abagana, Adam Yacoub; Zheng, Weiwei; Sun, Pengyu; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2017-04-01

    During an investigation exploring potential sources of novel species and natural products, a novel actinomycete with antifungal activity, designated strain NEAU-Gz11(T), was isolated from a soil sample, which was collected from Gama, Chad. The isolate was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain NEAU-Gz11(T) belongs to the genus Streptomyces with high sequence similarity to Streptomyces hiroshimensis JCM 4098(T) (98.0 %). Similarities to other type strains of the genus Streptomyces were lower than 98.0 %. However, the physiological and biochemical characteristics and low levels of DNA-DNA relatedness could differentiate the isolate genotypically and phenotypically from S. hiroshimensis JCM 4098(T). Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces gamaensis sp. nov. is proposed. The type strain is NEAU-Gz11(T) (=CGMCC 4.7304(T)=DSM 101531(T)).

  5. Streptomyces castaneus sp. nov., a novel actinomycete isolated from the rhizosphere of Peucedanum praeruptorum Dunn.

    PubMed

    Zhou, Shuyu; Li, Zhilei; Bai, Lu; Yan, Kai; Zhao, Junwei; Lu, Chang; Liu, Chongxi; Wang, Xiangjing; Xiang, Wensheng

    2017-01-01

    During an investigation of microbial diversity in medicinal herbs, a novel actinomycete, strain NEAU-QHHV11(T) was isolated from the rhizosphere of Peucedanum praeruptorum Dunn collected from Xianglu Mountain in Heilongjiang Province, northeast China and characterized using a polyphasic approach. The organism was found to have typical characteristics of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequence also indicated that strain NEAU-QHHV11(T) belongs to the genus Streptomyces and was most closely related to Streptomyces graminilatus NBRC 108882(T) (98.7 % sequence similarity) and Streptomyces turgidiscabies NBRC 16080(T) (98.7 % sequence similarity). The results of DNA-DNA hybridization and some phenotypic characteristics indicated that strain NEAU-QHHV11(T) could be distinguished from its close phylogenetic relatives. Thus, strain NEAU-QHHV11(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces castaneus sp. nov. is proposed. The type strain is NEAU-QHHV11(T) (=CGMCC 4.7235(T) = DSM 100520(T)).

  6. Streptomyces atlanticus sp. nov., a novel actinomycete isolated from marine sponge Aplysina fulva (Pallas, 1766).

    PubMed

    Silva, Fábio Sérgio Paulino; Souza, Danilo Tosta; Zucchi, Tiago Domingues; Pansa, Camila Cristiane; de Figueiredo Vasconcellos, Rafael Leandro; Crevelin, Eduardo José; de Moraes, Luiz Alberto Beraldo; Melo, Itamar Soares

    2016-11-01

    The taxonomic position of a novel marine actinomycete isolated from a marine sponge, Aplysina fulva, which had been collected in the Archipelago of Saint Peter and Saint Paul (Equatorial Atlantic Ocean), was determined by using a polyphasic approach. The organism showed a combination of morphological and chemotaxonomic characteristics consistent with its classification in the genus Streptomyces and forms a distinct branch within the Streptomyces somaliensis 16S rRNA gene tree subclade. It is closely related to Streptomyces violascens ISP 5183(T) (97.27 % 16S rRNA gene sequence similarity) and Streptomyces hydrogenans NBRC 13475(T) (97.15 % 16S rRNA gene sequence similarity). The 16S rRNA gene similarities between the isolate and the remaining members of the subclade are lower than 96.77 %. The organism can be distinguished readily from other members of the S. violacens subclade using a combination of phenotypic properties. On the basis of these results, it is proposed that isolate 103(T) (=NRRL B-65309(T) = CMAA 1378(T)) merits recognition as the type strain of a new Streptomyces species, namely Streptomyces atlanticus sp. nov.

  7. Streptomyces formicae sp. nov., a novel actinomycete isolated from the head of Camponotus japonicus Mayr.

    PubMed

    Bai, Lu; Liu, Chongxi; Guo, Lifeng; Piao, Chenyu; Li, Zhilei; Li, Jiansong; Jia, Feiyu; Wang, Xiangjing; Xiang, Wensheng

    2016-02-01

    During a screening for novel and biotechnologically useful actinobacteria in insects, a novel actinomycete with antifungal activity, designated strain 1H-GS9(T), was isolated from the head of a Camponotus japonicus Mayr ant, which were collected from Northeast Agricultural University (Harbin, Heilongjiang, China). Strain 1H-GS9(T) was characterised using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain 1H-GS9(T) belongs to the genus Streptomyces with high sequence similarities to Streptomyces scopuliridis DSM 41917(T) (98.8 %) and Streptomyces mauvecolor JCM 5002(T) (98.6 %). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it forms a monophyletic clade with Streptomyces kurssanovii JCM 4388(T) (98.6 %), Streptomyces xantholiticus JCM 4282(T) (98.6 %) and Streptomyces peucetius JCM 9920(T) (98.5 %). Thus, a combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 1H-GS9(T) and the above-mentioned five strains, which further clarified their relatedness and demonstrated that strain 1H-GS9(T) could be distinguished from these strains. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces formicae sp. nov. is proposed. The type strain is 1H-GS9(T) (=CGMCC 4.7277(T) = DSM 100524(T)).

  8. Streptomyces bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Jin, Pinjiao; Liu, Chongxi; Ma, Zhaoxu; Zhao, Junwei; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng

    2016-09-01

    A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).

  9. Cloning and characterization of the first actinomycete β-propeller phytase from Streptomyces sp. US42.

    PubMed

    Boukhris, Ines; Farhat-Khemakhem, Ameny; Bouchaala, Kameleddine; Virolle, Marie-Joëlle; Chouayekh, Hichem

    2016-10-01

    A gene encoding an extracellular phytase was cloned for the first time from an Actinomycete, Streptomyces sp. US42 and sequenced. The sequence of this gene revealed an encoded polypeptide (PHY US42) exhibiting one and six residues difference with the putative phytases of Streptomyces lividans TK24 and Streptomyces coelicolor A3(2), respectively. The molecular modeling of PHY US42 indicated that this phytase belongs to the group of β-propeller phytases that are usually calcium-dependent. PHY US42 was purified and characterized. Its activity was calcium-dependent and maximal at pH 7 and 65 °C. The enzyme was perfectly stable at pH ranging from 5 to 10 and its thermostability was greatly enhanced in the presence of calcium. Indeed, PHY US42 maintained 80% of activity after 10 min of incubation at 75 °C in the presence of 5 mM CaCl2 . PHY US42 was also found to exhibit high stability after incubation at 37 °C for 1 h in the presence of bovine bile and digestive proteases like of pepsin, trypsin, and chymotrypsin. Considering its biochemical properties, PHY US42 could be used as feed additive in combination with an acid phytase for monogastric animals.

  10. Streptomyces luozhongensis sp. nov., a novel actinomycete with antifungal activity and antibacterial activity.

    PubMed

    Zhang, Renwen; Han, Xiaoxue; Xia, Zhanfeng; Luo, Xiaoxia; Wan, Chuanxing; Zhang, Lili

    2017-02-01

    A novel actinomycete strain, designated TRM 49605(T), was isolated from a desert soil sample from Lop Nur, Xinjiang, north-west China, and characterised using a polyphasic taxonomic approach. The strain exhibited antifungal activity against the following strains: Saccharomyces cerevisiae, Curvularia lunata, Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Penicillium citrinum, Candida albicans and Candida tropicalis; Antibacterial activity against Bacillus subtilis, Staphylococcus epidermidis and Micrococcus luteus; and no antibacterial activity against Escherichia coli. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 49605(T) to the genus Streptomyces. Strain TRM 49605(T) shows high sequence similarities to Streptomyces roseolilacinus NBRC 12815(T) (98.62 %), Streptomyces flavovariabilis NRRL B-16367(T) (98.45 %) and Streptomyces variegatus NRRL B-16380(T) (98.45 %). Whole cell hydrolysates of strain TRM 49605(T) were found to contain LL-diaminopimelic acid as the diagnostic diamino acid and galactose, glucose, xylose and mannose as the major whole cell sugars. The major fatty acids in strain TRM 49605(T) were identified as iso C16:0, anteiso C15:0, C16:0 and Summed Feature 5 as defined by MIDI. The main menaquinones were identified as MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The G+C content of the genomic DNA was determined to be 71.2 %. The DNA-DNA relatedness between strain TRM 49605(T) and the phylogenetically related strain S. roseolilacinus NBRC 12815(T) was 60.12 ± 0.06 %, which is lower than the 70 % threshold value for delineation of genomic prokaryotic species. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain TRM 49605(T) (=CCTCC AA2015026(T) = KCTC 39666(T)) should be designated as the type strain of a novel species of the genus

  11. Tartrolon D, a cytotoxic macrodiolide from the marine-derived actinomycete Streptomyces sp. MDG-04-17-069.

    PubMed

    Pérez, Marta; Crespo, Cristina; Schleissner, Carmen; Rodríguez, Pilar; Zúñiga, Paz; Reyes, Fernando

    2009-12-01

    Exploration of marine-derived actinomycetes as a source of antitumor compounds has led to the isolation of a new member of the tartrolon series, tartrolon D (4). This new compound was obtained from Streptomyces sp. MDG-04-17-069 fermentation broths and displayed strong cytotoxic activity against three human tumor cell lines. Additionally, the known compound ikarugamycin (5) was also found in the culture broths of the same microorganism. The structure of this new tartrolon was established by a combination of spectroscopic techniques (1D and 2D NMR, HRMS, and UV) as well as by comparison with published data for similar compounds.

  12. Streptomyces xinjiangensis sp. nov., an actinomycete isolated from Lop Nur region.

    PubMed

    Cheng, Cong; Li, Yu-Qian; Asem, Mipeshwaree Devi; Lu, Chun-Yan; Shi, Xiao-Han; Chu, Xiao; Zhang, Wan-Qin; Di An, Deng-; Li, Wen-Jun

    2016-10-01

    A novel actinobacterial strain, designated LPA192(T), was isolated from a soil sample collected from Lop Nur, Xinjiang Uygur Autonomous Region, Northwest China. A polyphasic approach was used to investigate the taxonomic position of strain LPA192(T). The isolate showed morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Peptidoglycan was found to contain LL-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinones were MK-9(H6) and MK-10(H4). Polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol. Major cellular fatty acids consist of C16:0, anteiso-C15:0 and C18:1 ω9c. The sugar in whole-cell hydrolysates was mannose. Phylogenetic analysis indicated that strain LPA192(T) is closely related to Streptomyces tanashiensis LMG 20274(T) (99.3 %), Streptomyces gulbargensis DAS131(T) (99.3 %), Streptomyces nashvillensis NBRC 13064(T) (99.3 %), Streptomyces roseolus NBRC 12816(T) (99.2 %) and Streptomyces filamentosus NBRC 12767(T) (99.1 %) while showing below 98.5 % sequencing similarities with other validly published Streptomyces species. However, DNA-DNA relatedness values between LPA192(T) and the closely related type strains were below 40 %, which are much lower than 70 % threshold value for species delineation. The genomic DNA G + C content of strain LPA192(T) was 69.3 mol %. Based on the differences in genotypic and phenotypic characteristics from the closely related strains, strain LPA192(T) is considered to represent a novel species of the genus Streptomyces for which the name Streptomyces xinjiangensis sp. nov. is proposed. The type strain is LPA192(T) (=KCTC 39601(T) = CGMCC 4.7288(T)).

  13. Nematicidal activity of fervenulin isolated from a nematicidal actinomycete, Streptomyces sp. CMU-MH021, on Meloidogyne incognita.

    PubMed

    Ruanpanun, Pornthip; Laatsch, Hartmut; Tangchitsomkid, Nuchanart; Lumyong, Saisamorn

    2011-06-01

    An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.

  14. Streptomyces manipurensis sp. nov., a novel actinomycete isolated from a limestone deposit site in Manipur, India.

    PubMed

    Nimaichand, Salam; Zhu, Wen-Yong; Yang, Ling-Ling; Ming, Hong; Nie, Guo-Xing; Tang, Shu-Kun; Ningthoujam, Debananda S; Li, Wen-Jun

    2012-06-01

    A novel actinobacterium, designated MBRL 201(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. The strain was characterized using polyphasic taxonomy. Comparison of the 16S rRNA gene sequence of strain MBRL 201(T) and other Streptomyces species showed sequence similarities ranging from 93.0 to 99.6 % and strain MBRL 201(T) showed closest similarities to Streptomyces virginiae NBRC 12827(T) (99.6 %) and Streptomyces cinnamonensis NBRC 15873(T) (99.6 %). The DNA relatedness between MBRL 201(T) and the type strains of S. virginiae NBRC 12827(T) and S. cinnamonensis NBRC 15873(T) were 44.5 and 35.6 % respectively. Strain MBRL 201(T) contained LL: -diaminopimelic acid (A(2)pm) as the diagnostic diamino acid, with glucose as the main sugar, while small amounts of galactose, glucose, mannose, rhamnose, ribose and xylose were also present in cell-wall hydrolysates. The major fatty acids identified were anteiso-C(15:0) (38.9 %), iso-C(15:0) (19.9 %) and anteiso-C(17:1) (14.7 %). The predominant menaquinones detected were MK-9(H(6)) and MK-9(H(8)), while the polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides, with other unknown phospholipids and lipids. The G+C content of the genomic DNA was 72.9 %. The phenotypic and genotypic data showed that strain MBRL 201(T) merits recognition as a representative of a novel species of the genus Streptomyces. It is proposed that the isolate should be classified in the genus Streptomyces as a novel species, Streptomyces manipurensis sp. nov. The type strain is MBRL 201(T) (=DSM 42029(T) = JCM 17351(T)).

  15. Streptomyces songpinggouensis sp. nov., a Novel Actinomycete Isolated from Soil in Sichuan, China.

    PubMed

    Guan, Xuejiao; Li, Wenchao; Liu, Chongxi; Jin, Pinjiao; Guo, Siyu; Wang, Xiangjing; Xiang, Wensheng

    2016-12-01

    During a screening for novel and biotechnologically useful actinobacteria, a novel actinobacteria with weak antifungal activity, designated strain NEAU-Spg19(T), was isolated from a soil sample collected from pine forest in Songpinggou, Sichuan, southwest China. The strain was characterized using a polyphasic taxonomic approach which confirmed that it belongs to the genus Streptomyces. Growth occurred at a temperature range of 10-30 °C, pH 5.0-11.0 and NaCl concentrations of 0-5 %. The cell wall peptidoglycan consisted of LL-diaminopimelic acid and glycine. The major menaquinones were MK-9(H6), MK-9(H8) and MK-9(H4). The phospholipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine and phosphatidylinositol. The major fatty acids were iso-C15:0, iso-C16:0, and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-Spg19(T) belongs to the genus Streptomyces with the highest sequence similarities to Streptomyces tauricus JCM 4837(T) (98.6 %) and Streptomyces rectiviolaceus JCM 9092(T) (98.3 %). Some physiological and biochemical properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. tauricus JCM 4837(T) and S. rectiviolaceus JCM 9092(T). Hence, on the basis of phenotypic and genetic analyses, it is proposed that strain NEAU-Spg19(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces songpinggouensis sp. nov. is proposed. The type strain is NEAU-Spg19(T) (=CGMCC 4.7140(T)=DSM 42141(T)).

  16. Antimicrobial potential of phylogenetically unique actinomycete, Streptomyces sp. JRG-04 from marine origin.

    PubMed

    Govindarajan, Ganesan; Satheeja Santhi, Velayudhan; Jebakumar, Solomon Robinson David

    2014-11-01

    Due to the emergence of severe infectious diseases and thriving antibiotic resistance, there is a need to explore microbial-derived bioactive secondary metabolites from unexplored regions. Present study deals with a mangrove estuary derived strain of Streptomyces sp. with potent antimicrobial activity against various pathogens, including methicillin resistant Staphylococcus aureus. Bioactive compound was effective even at low MIC level, damages the membrane of methicillin resistant S. aureus and causes cell death, however it has no cytotoxic effect on H9C2 cells. 16S rRNA shared 99.5% sequence similarity to Streptomyces longispororuber. Optimum biomass and antimicrobial compound production were observed in production medium supplemented with 1.0% maltose and 0.5% yeast extract. The active compound purified from the chloroform extract of the cell-free supernatant was studied by FT-IR, 1H NMR, 13C NMR and LC ESI-MS and identified as aromatic polyketide. β-ketosynthase (KS) domain of the Streptomyces strain revealed 93.2% sequence similarity to the benzoisochromanequinone, an actinorhodin biosynthetic gene cluster of Streptomyces coelicolor A3(2). However, the region synthesizing the secondary metabolite produced by the S. longispororuber was not related to the KS domain of the strain, due to the phenomenon of horizontal gene transfer over the period of evolutionary process, thus generating metabolic compound diversity.

  17. Comparative analysis of chemical constituents, antimicrobial and antioxidant activities of ethylacetate extracts of Polygonum cuspidatum and its endophytic actinomycete, Streptomyces sp. A0916.

    PubMed

    Wang, Lei; Qiu, Peng; Long, Xiu-Feng; Zhang, Shuai; Zeng, Zhi-Gang; Tian, Yong-Qiang

    2016-02-01

    The present study investigated the chemical composition of ethylacetate extracts from an endophytic actinomycete Streptomyces sp. A0916 and its host Polygonum cuspidatum. A comparative analysis of the antimicrobial and antioxidant properties of the extracts was also conducted. 32 compounds of P. cuspidatum and 23 compounds of Streptomyces sp. A0916 were isolated and identified by GC/MS. Antimicrobial activities of the extracts were evaluated using eight microbial strains (3 Gram-positive bacteria, 3 Gram-negative bacteria, and 2 fungi). The Streptomyces sp. A0916 extracts showed a wide range of antimicrobial activities and presented greater antimicrobial effectiveness than the P. cuspidatum extracts. The minimum inhibitory concentration (MIC) of Streptomyces sp. A0916 extracts against the ampicillin-resistant strain Enterococcus faecium SIIA843 was 32 μg·mL(-1). Furthermore, the extracts had greater antimicrobial effect against Gram-positive bacteria than Gram-negative bacteria. Finally, the antioxidant activity of the Streptomyces sp. A0916 extracts was equal to that of the P. cuspidatum extracts. In conclusion, our results suggest that the endophytic actinomycetes of the medicinal plants are an important source of bioactive substances.

  18. Streptomyces halophytocola sp. nov., an endophytic actinomycete isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour.

    PubMed

    Qin, Sheng; Bian, Guang-Kai; Tamura, Tomohiko; Zhang, Yue-Ji; Zhang, Wen-Di; Cao, Cheng-Liang; Jiang, Ji-Hong

    2013-08-01

    A novel actinomycete, designated KLBMP 1284(T), was isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour. collected from the city of Nantong, Jiangsu Province, east China. The strain was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Analysis of the 16S rRNA gene sequence of strain KLBMP 1284(T) revealed that the strain formed a distinct clade within the phylogenetic tree based on 16S rRNA gene sequences and the highest sequence similarity (99.43 %) was to Streptomyces sulphureus NRRL B-1627(T). 16S rRNA gene sequence similarity to other species of the genus Streptomyces was lower than 97 %. Based on DNA-DNA hybridization values and comparison of morphological and phenotypic data, KLBMP 1284(T) could be distinguished from the closest phylogenetically related species, Streptomyces sulphureus NRRL B-1627(T). Thus, based on these data, it is evident that strain KLBMP 1284(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces halophytocola sp. nov. is proposed. The type strain is KLBMP 1284(T) (= KCTC 19890(T) = NBRC 108770(T)).

  19. Cephamycins, a New Family of β-Lactam Antibiotics I. Production by Actinomycetes, Including Streptomyces lactamdurans sp. n1

    PubMed Central

    Stapley, E. O.; Jackson, M.; Hernandez, S.; Zimmerman, S. B.; Currie, S. A.; Mochales, S.; Mata, J. M.; Woodruff, H. B.; Hendlin, D.

    1972-01-01

    A number of actinomycetes isolated from soil were found to produce one or more members of a new family of antibiotics, the cephamycins, which are structurally related to cephalosporin C. The cephamycins were produced in submerged fermentation in a wide variety of media by one or more of eight different species of Streptomyces, including a newly described species, S. lactamdurans. These antibiotics exhibit antibacterial activity against a broad spectrum of bacteria which includes many that are resistant to the cephalosporins and penicillins. PMID:4790552

  20. Streptomyces tritolerans sp. nov., a novel actinomycete isolated from soil in Karnataka, India.

    PubMed

    Syed, Dastager G; Agasar, Dayanand; Kim, Chang-Jin; Li, Wen-Jun; Lee, Jae-Chan; Park, Dong-Jin; Xu, Li-Hua; Tian, Xin-Peng; Jiang, Cheng-Lin

    2007-11-01

    A Gram-positive, nonmotile, moderately halophilic, alkali and thermotolerant strain designated DAS 165(T), was isolated from a dry land soil sample from the Gulbarga region, Karnataka province, India. The isolate produced yellow substrate mycelia and gray aerial mycelia on most tested media. Strain DAS 165(T) showed growth in the presence of 5 to 7% NaCl and at 45 degrees C. The DNA G + C content was 69.7%. 16S rRNA gene sequence analysis together with these characteristics consistently assigned strain DAS 165(T) to the genus Streptomyces. The 16S rRNA gene sequence analysis revealed that strain DAS 165(T) was most closely related to S. tendae ATCC 19812(T) (D 63873) with a sequence similarity of 99.6% (three nucleotide differences out of 1,517). Strain DAS 165(T) formed a distinct clade based on analysis of the almost complete sequence and 120-nucleotide variable gamma region of the 16S rRNA gene. Despite the high sequence similarity, strain DAS 165(T) was phenotypically different from S. tendae ATCC 19812(T). DNA-DNA hybridization between these strains was 47% showing that strain DAS 165(T) is a distinct genomic species. Phenetic and genetic results support the classification of strain DAS 165(T) as a new species, for which the name S. tritolerans is proposed, with strain DAS 165(T) as the type strain (=DSM 41899(T )= CCTCCAA 206013(T)).

  1. Complete genome sequence of Streptomyces sp. strain CFMR 7, a natural rubber degrading actinomycete isolated from Penang, Malaysia.

    PubMed

    Nanthini, Jayaram; Chia, Kim-Hou; Thottathil, Gincy P; Taylor, Todd D; Kondo, Shinji; Najimudin, Nazalan; Baybayan, Primo; Singh, Siddharth; Sudesh, Kumar

    2015-11-20

    Streptomyces sp. strain CFMR 7, which naturally degrades rubber, was isolated from a rubber plantation. Whole genome sequencing and assembly resulted in 2 contigs with total genome size of 8.248 Mb. Two latex clearing protein (lcp) genes which are responsible for rubber degrading activities were identified.

  2. Streptomyces oceani sp. nov., a new obligate marine actinomycete isolated from a deep-sea sample of seep authigenic carbonate nodule in South China Sea.

    PubMed

    Tian, Xin-Peng; Xu, Ying; Zhang, Jing; Li, Jie; Chen, Zhong; Kim, Chang-Jin; Li, Wen-Jun; Zhang, Chang-Sheng; Zhang, Si

    2012-08-01

    A novel aerobic actinomycete strain, designated as SCSIO 02100(T), was isolated from a deep sea sediment sample collected from Northern South China Sea at a depth of 578 m. This isolate requires sea water or a sodium-supplemented medium for growth. BLAST searches based on the almost full length of the 16S rRNA gene sequence, showed that strain SCSIO 02100(T) had the highest similarities with Streptomyces armeniacus (JCM 3070(T)) (97.1 %). Phylogenetic trees reconstructed on the basis of 16S rRNA gene sequences revealed that strain SCSIO 02100(T) formed a distinct lineage with S. nanshensis SCSIO 01066(T) with 96.9 % similarity. Further analysis of the polyphasic taxonomic data, including morphological, phenotypic and chemotaxonomic properties, showed that strain SCSIO 02100(T) could be readily distinguished from the most closely related members of the genus Streptomyces. Thus, based on the polyphasic taxonomic data, a novel species, Streptomyces oceani sp. nov., is proposed, with the type strain SCSIO 02100(T) (=DSM 42043(T) = CGMCC 4.7007(T)).

  3. Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4

    PubMed Central

    Sudha, S; Masilamani, Selvam M

    2012-01-01

    Objective To investigate the cytotoxic activity of actinomycete isolated from marine sediment. Methods In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction, using two universal bacterial primers, 1492R (5′-GGTTACCTTGTTAC GACTT-3′) and Eubac27F (5′-AGAGTTTGATCCTGGCTC AG-3′). The amplified products were purified using TIANgel mini purification kit, ligated to MD18-T simple vector (TaKaRa), and transformed into competent cells of Escherichia coli DH5α. 16S rRNA gene fragment was sequenced using forward primer M13F (-47) and reverse primer M13R (-48). Blast search sequence similarity was found against the existing non-redundant nucleotide sequence database thus, identified as Streptomyces sp SU, Streptomyces rubralavandulae strain SU1, Streptomyces cacaoi strain SU2, Streptomyces cavourensis strain SU3, Streptomyces avidinii strain SU4, Streptomyces globisporus strain SU5, Streptomyces variabilis strain SU6, Streptomyces coelicolor strain SU 7. Among the eight identified isolates, one actinomycete Streptomyces avidinii strain SU4 was selected for further study. Results Crude extract of the actinomycete isolate exhibited IC50 in 64.5 µg against Hep-2 cell line, 250 µg in VERO cell line. This value is very close to the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI) is in IC50 < 30 µg/mL. The GC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (12.17%), isooctyl phthalate (15.29%) with the retention time 15.642 and 21.612, respectively. Conclusions This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post

  4. Streptomyces chlorus sp. nov. and Streptomyces viridis sp. nov., isolated from soil.

    PubMed

    Kim, Byung-Yong; Rong, Xiaoying; Zucchi, Tiago D; Huang, Ying; Goodfellow, Michael

    2013-05-01

    Two actinomycete strains, BK125(T) and BK199(T), isolated from a hay meadow soil sample were investigated to determine their taxonomic position using a polyphasic approach. The isolates produced greenish-yellow and light green aerial mycelium on oatmeal agar, respectively. They contained anteiso-C15 : 0, iso-C15 : 0 and C16 : 0 as the major fatty acids, and MK-9 (H6) and MK-9 (H8) as the predominant isoprenoid quinones. Phylogenetic analysis of the 16S rRNA gene sequences showed that the isolates formed distinct phyletic lines towards the periphery of the Streptomyces prasinus subclade. Analysis of DNA-DNA relatedness between the two isolates showed that they belonged to different genomic species. The organisms were also distinguished from one another and from type strains of species classified in the S. prasinus subclade using a combination of genotypic and phenotypic properties. On the basis of these data, it is proposed that the isolates be assigned to the genus Streptomyces as Streptomyces chlorus sp. nov. and Streptomyces viridis sp. nov. with isolates BK125(T) ( = KACC 20902(T) = CGMCC 4.5798(T)) and BK199(T) ( = KACC 21003(T) = CGMCC 4.6824(T)) as the respective type strains.

  5. Streptomyces hyaluromycini sp. nov., isolated from a tunicate (Molgula manhattensis).

    PubMed

    Harunari, Enjuro; Hamada, Moriyuki; Shibata, Chiyo; Tamura, Tomohiko; Komaki, Hisayuki; Imada, Chiaki; Igarashi, Yasuhiro

    2016-03-01

    A novel Gram-stain-positive actinomycete, designated MB-PO13(T), was isolated from a tunicate (Molgula manhattensis) collected in Tokyo Bay, Japan, and its taxonomic position was studied by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain MB-PO13(T) was closely related to Streptomyces graminisoli JR-12(T) (99.72% 16S rRNA gene sequence similarity) and Streptomyces shenzhenensis 172115(T) (99.23%). The strain contained LL-diaminopimelic acid in the whole-cell hydrolysate. The predominant menaquinones were MK-9(H8) and MK-9(H6) and the major fatty acids were anteiso-C15:0, iso-C16:0, iso-C14:0 and C16:0. These data supported the affiliation of the novel strain to the genus Streptomyces. Meanwhile, results of DNA-DNA hybridization and physiological and biochemical tests indicated that strain MB-PO13(T) was distinguished from known Streptomyces type strains. Therefore, strain MB-PO13(T) represents a novel species of the genus Streptomyces for which the name Streptomyces hyaluromycini sp. nov. is proposed; the type strain is MB-PO13(T) (=NBRC 110483(T) =DSM 100105(T)).

  6. New streptophenazines from marine Streptomyces sp. 182SMLY.

    PubMed

    Liang, Ying; Chen, Lu; Ye, Xuewei; Anjum, Komal; Lian, Xiao-Yuan; Zhang, Zhizhen

    2017-02-01

    Six phenazines including three new ones were isolated from the culture of a marine actinomycete Streptomyces sp. 182SMLY. Based on the analyses of NMR, HRESIMS, optical rotation value, and CD data, the structures of these isolated compounds were determined as new phenazines of (-)-streptophenazines M-O and known phenazines of 1-carbomethoxyphenazine and (-)-streptophenazines A and B. (-)-Streptophenazine B showed activity in suppressing the growth of methicillin-resistant Staphylococcus aureus with MIC value of 4.2 μg/mL.

  7. Genetic manipulation of secondary metabolite biosynthesis for improved production in Streptomyces and other actinomycetes.

    PubMed

    Baltz, Richard H

    2016-03-01

    Actinomycetes continue to be important sources for the discovery of secondary metabolites for applications in human medicine, animal health, and crop protection. With the maturation of actinomycete genome mining as a robust approach to identify new and novel cryptic secondary metabolite gene clusters, it is critical to continue developing methods to activate and enhance secondary metabolite biosynthesis for discovery, development, and large-scale manufacturing. This review covers recent reports on promising new approaches and further validations or technical improvements of existing approaches to strain improvement applicable to a wide range of Streptomyces species and other actinomycetes.

  8. Genome Sequence of Streptomyces viridosporus Strain T7A ATCC 39115, a Lignin-Degrading Actinomycete

    PubMed Central

    Davis, Jennifer R.; Goodwin, Lynne; Teshima, Hazuki; Detter, Chris; Tapia, Roxanne; Han, Cliff; Huntemann, Marcel; Wei, Chia-Lin; Han, James; Chen, Amy; Kyrpides, Nikos; Mavrommatis, Kostas; Szeto, Ernest; Markowitz, Victor; Ivanova, Natalia; Mikhailova, Natalia; Ovchinnikova, Galina; Pagani, Ioanna; Pati, Amrita; Woyke, Tanja; Pitluck, Sam; Peters, Lin; Nolan, Matt; Land, Miriam

    2013-01-01

    We announce the availability of the genome sequence of Streptomyces viridosporus strain T7A ATCC 39115, a plant biomass-degrading actinomycete. This bacterium is of special interest because of its capacity to degrade lignin, an underutilized component of plants in the context of bioenergy. It has a full complement of genes for plant biomass catabolism. PMID:23833133

  9. Genome Sequence of Streptomyces viridosporus Strain T7A ATCC 39115, a Lignin-Degrading Actinomycete

    SciTech Connect

    Davis, Jennifer R.; Goodwin, Lynne A.; Teshima, Hazuki; Detter, J. Chris; Tapia, Roxanne; Han, Cliff; Huntemann, Marcel; Wei, Chia-Lin; Han, James; Chen, Amy; Kyrpides, Nikos C; Mavromatis, K; Szeto, Ernest; Markowitz, Victor; Ivanova, N; Mikhailova, Natalia; Ovchinnikova, Galina; Pagani, Ioanna; Pati, Amrita; Woyke, Tanja; Pitluck, Sam; Peters, Lin; Nolan, Matt; Land, Miriam L; Sello, Jason K.

    2013-01-01

    We announce the availability of the genome sequence of Streptomyces viridosporus strain T7A ATCC 39115, a plant biomass- degrading actinomycete. This bacterium is of special interest because of its capacity to degrade lignin, an underutilized compo- nent of plants in the context of bioenergy. It has a full complement of genes for plant biomass catabolism.

  10. Friedmanniella flava sp. nov., a soil actinomycete.

    PubMed

    Zhang, Xuefang; Zhang, Jianli; Zhang, Yabo; Xin, Yuhua; He, Hongju

    2013-05-01

    A novel actinomycete, strain W6(T), was isolated from a soil sample of Yunnan Province, China. The bacterium was aerobic, non-motile, non-spore-forming and Gram-stain-positive. Genetic, phenotypic and chemical properties of the isolate were studied. 16S rRNA gene sequence data suggested that the novel isolate belonged to the genus Friedmanniella and shared 98.6% sequence similarity with Friedmanniella antarctica DSM 11053(T) and Friedmanniella okinawensis DSM 21744(T), the most closely related species. The cell-wall peptidoglycan contained ll-diaminopimelic acid, and mycolic acids were absent. The main menaquinone was MK-9(H4) and the predominant fatty acids were anteiso-C15:0 and iso-C15:0. The phospholipid profile contained phosphatidylglycerol, phosphatidylinositol, phosphatidylcholine and diphosphatidylglycerol. The DNA G+C content of strain W6(T) was 72 mol%. Strain W6(T) showed 30.0% and 28.5% DNA-DNA relatedness, respectively, to F. antarctica DSM 11053(T) and F. okinawensis DSM 21744(T). The combined genotypic and phenotypic data showed that strain W6(T) should be assigned to the genus Friedmanniella as a representative of a novel species, for which the name Friedmanniella flava sp. nov. is proposed. The type strain is W6(T) ( = CGMCC 4.6856(T) =JCM 17701(T)).

  11. Isolation and identification of biocontrol agent Streptomyces rimosus M527 against Fusarium oxysporum f. sp. cucumerinum.

    PubMed

    Lu, Dandan; Ma, Zheng; Xu, Xianhao; Yu, Xiaoping

    2016-08-01

    Actinomycetes have received considerable attention as biocontrol agents against fungal plant pathogens and as plant growth promoters. In this study, a total of 320 actinomycetes were isolated from various habitats in China. Among which, 77 strains have been identified as antagonistic activities against Fusarium oxysporum f. sp. cucumerinum which usually caused fusarium wilt of cucumber. Of these, isolate actinomycete M527 not only displayed broad-spectrum antifungal activity but also showed the strongest antagonistic activity against the spore germination of F. oxysporum f. sp. cucumerinum. In pot experiments, the results indicated that isolate M527 could promote the shoot growth and prevent the development of the disease on cucumber caused by F. oxysporum f. sp. cucumerinum. The control efficacy against seedling fusarium wilt of cucumber after M527 fermentation broth root-irrigation was up to 72.1% as compared to control. Based on 16S rDNA sequence analysis, the isolate M527 was identified as Streptomyces rimosus.

  12. Chromate reductase activity in Streptomyces sp. MC1.

    PubMed

    Polti, Marta A; Amoroso, María J; Abate, Carlos M

    2010-02-01

    Biological transformation of Cr(VI) to Cr(III) by enzymatic reduction may provide a less costly and more environmentally friendly approach to remediation. In a previous report a Cr(VI) resistant actinomycete strain, Streptomyces sp. MC1, was able to reduce Cr(VI) present in a synthetic medium, soil extract and soil samples. This is the first time optimal conditions such as pH, temperature, growth phase and electron donor have been elucidated in vitro for Cr(VI) reduction by a streptomycete. Chromate reductase of Streptomyces sp. MC1 is a constitutive enzyme which was mainly associated with biomass and required NAD(P)H as an electron donor. It was active over a broad temperature (19-39 degrees C) and pH (5-8) range, and optimum conditions were 30 degrees C and pH 7. The enzyme was present in supernatant, pellet and cell free extract. Bioremediation with the enzyme was observed in non-compatible cell reproduction systems, conditions frequently found in contaminated environments.

  13. Bioremediation of chromium(VI) contaminated soil by Streptomyces sp. MC1.

    PubMed

    Polti, Marta A; García, Roberto O; Amoroso, María J; Abate, Carlos M

    2009-06-01

    This work provides quantitative information on Cr(VI) reduction in soil samples by an indigenous actinomycete. Streptomyces sp. MC1, previously isolated from sugarcane, has shown ability to reduce Cr(VI) in liquid minimal medium. A reduction of 100 and 75% was obtained at initial Cr(VI) concentrations of 5 and 50 mg l(-1), respectively, after 48 h of incubation. Bioremediation ability of Streptomyces sp. MC1 was assayed in soil extracts and soil samples. Relative growth of Streptomyces sp. MC1 was 77 and 38% when grown in soil extract with 10 and 50 mg l(-1) of Cr(VI), respectively. MC1 was able to reduce 30% of Cr(VI) after 96 h of incubation with 10 mg l(-1) of Cr(VI), and reduction coincided with the exponential growth phase at pH 7 and 30 degrees C.In soil samples, Streptomyces sp. MC1 was able to reduce up to 94% of the Cr(VI) bioavailability (50 mg kg(-1)) after 7 d. These results were compared with non-inoculated soil samples with Cr(VI). Bioremediation activity of Streptomyces sp. MC1 was not inhibited by natural soil microbial flora. Besides, Streptomyces sp. MC1 growth was not inhibited by 50 mg kg(-1) of Cr(VI). In contrast to findings obtained by other authors, our results showed almost complete Cr(VI) removal from soil without any previous treatment, and without addition of any substrate and with a normal soil humidity level. These results confirm the Cr(VI)-contaminated soil bioremediation potential of Streptomyces sp. MC1.

  14. Immunologic relatedness of extracellular ligninases from the actinomycetes Streptomyces viridosporus T7A and Streptomyces badius 252

    SciTech Connect

    Magnuson, T.S.; Roberts, M.A.; Crawford, D.L.; Hertel, G.

    1991-12-31

    Four isoforms of the extracellular lignin peroxidase of the ligninolytic actinomycete Streptomyces viridosporus T7A (ALip-P1, P2, P3, and P4) were individually purified by ultrafiltration and ammonium sulfate precipitation, followed by electro-elution using polyacrylamide gel electrophoresis. Three of the purified peroxidases were compared for their immunologic relatedness by Western blot analysis using a polyclonal antibody preparation produced in rabbits against pure isoform P3. The anti-P3 antibody was also tested for its reactivity towards a lignin peroxidase from the white-rot fungus Phanerochaete chrysosporium and another ligninolytic actinomycete Streptomyces badius 252. Results showed that peroxidases ALip-P1 through ALip-P3 are immunologically related to one another. The peroxidases of S. badius, but not the peroxidase of P. chrysosporium, also reacted with the antibody, thus indicating that the lignin peroxidases of S. viridosporus and S. badius are immunologically related. Based upon its specific affinity, fignin peroxidase isoform ALip-P3 of S. viridosporus was readily purified using an anti-P3 antibody affinity column.

  15. Streptomyces palmae sp. nov., isolated from oil palm (Elaeis guineensis) rhizosphere soil.

    PubMed

    Sujarit, Kanaporn; Kudo, Takuji; Ohkuma, Moriya; Pathom-Aree, Wasu; Lumyong, Saisamorn

    2016-10-01

    Actinomycete strain CMU-AB204T was isolated from oil palm rhizosphere soil collected in Chiang Mai University (Chiang Mai, Thailand). Based on morphological and chemotaxonomic characteristics, the organism was considered to belong to the genus Streptomyces. Whole cell-wall hydrolysates consisted of ll-diaminopimelic acid, glucose, ribose and galactose. The predominant menaquinones were MK-9(H4), MK-9(H6), MK-9(H2) and MK-8(H4). The fatty acid profile contained iso-C15 : 0, iso-C16 : 0 and anteiso-C15 : 0 as major components. The principal phospholipids detected were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The DNA G+C content of strain CMU-AB204T was 70.9 mol%. Based on 16S rRNA gene sequence similarity, strain CMU-AB204T was closely related to Streptomyces orinoci JCM 4546T (98.7 %), Streptomyces lilacinus NBRC 12884T (98.5 %), Streptomyces abikoensis CGMCC 4.1662T (98.5 %), Streptomyces griseocarneus JCM 4905T (98.4 %) and Streptomyces xinghaiensis JCM 16958T (98.3 %). Phylogenetic trees revealed that the new strain had a distinct taxonomic position from closely related type strains of the genus Streptomyces. Spiny to hairy spores clearly differentiated strain CMU-AB204T from the five most closely related Streptomyces species, which produced smooth spores. On the basis of evidence from this polyphasic study, it is proposed that strain CMU-AB204T represents a novel species of the genus Streptomyces, namely Streptomyces palmae sp. nov. The type strain is CMU-AB204T (=JCM 31289T=TBRC 1999T).

  16. Extraction and Identification of Antibacterial Secondary Metabolites from Marine Streptomyces sp. VITBRK2

    PubMed Central

    Rajan, Benita Mercy; Kannabiran, Krishnan

    2014-01-01

    Actinomycetes were isolated from marine sediment samples collected from the east coast of Chennai, Tamil Nadu, India. Well diffusion and agar plug methods were used for the evaluation of antibiotic production by these isolates against drug resistant Methicillin- resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococci (VRE). The potential isolate VITBRK2 was mass cultured for morphological and physiological characterization. The culturing conditions of the isolate were optimized and the recommendations of International Streptomyces Project were followed for the assimilation of carbon and nitrogen sources. The isolate was identified by comparing the properties with representative species in the key of Nonomura and Bergey’s Manual of Determinative Bacteriology. Ethyl acetate extract prepared from the cell free culture broth of the isolate was analyzed using HPLC- diode array technique to characterize the metabolites and identify the antibiotics. VITBRK2 was found to be Gram-positive rod grey color aerial mycelium production. It was also non motile in nature with spiral spore chain morphology. VITBRK2 was identified as Streptomyces and designated as Streptomyces sp. VITBRK2. HPLC-DAD analysis showed the presence of indolo compounds (3- methyl-indole and 2-methyl- indole) along with amicoumacin antibiotic. The observed activity of Streptomyces sp. VITBRK2 against MRSA and VRE strains may be due to the presence of indolo compounds in the isolate. The results of this study suggested that secondary metabolites produced by Streptomyces sp. VITBRK2 could be used as a lead to control drug resistant bacterial pathogens. PMID:25317399

  17. Cadmium biosorption by Streptomyces sp. F4 isolated from former uranium mine.

    PubMed

    Siñeriz, Manuel Louis; Kothe, Erika; Abate, Carlos Mauricio

    2009-09-01

    46 actinomycetes were isolated from two polluted sites and one unpolluted site. One strain, F4, was selected through primary qualitative screening assays because of its cadmium resistance, and physiologically and taxonomically characterized. F4 was able to grow at 7.5% NaCl and 100 microg/ml lysozyme and at a pH between 6 and 10. 16S rDNA sequence analysis showed that F4 was closely related to Streptomyces tendae. Growth of Streptomyces sp. F4 on culture medium with 8 mg/l Cd(2+) for 8 days showed 80% inhibition. Maximum specific biosorption was 41.7 mg Cd(2+)/g dry weight after 7 days of growth and highest Cd(2+ )concentration was found in the cell wall (41.2%). The exopolysaccharide layer only contained 7.4%, whereas 39.4% of Cd(2+) was found in the cytosolic fraction. Twelve % was found in the ribosomes and membrane fraction. This was verified with TEM, showing Streptomyces sp. F4 cytoplasm with dark granulate appearance. This study could present the potential capacity of Streptomyces sp. F4 for Cd(2+) bioremediation.

  18. Streptomyces actinomycinicus sp. nov., isolated from soil of a peat swamp forest.

    PubMed

    Tanasupawat, Somboon; Phongsopitanun, Wongsakorn; Suwanborirux, Khanit; Ohkuma, Moriya; Kudo, Takuji

    2016-01-01

    A novel actinomycete, strain RCU-197T, was isolated from soil of a peat swamp forest in Rayong Province, Thailand. Using a polyphasic approach, the strain was classified in the genus Streptomyces. It contained ll-diaminopimelic acid in the cell-wall peptidoglycan. No diagnostic sugars were detected in whole-cell hydrolysates and there was a lack of mycolic acids. The major menaquinones were MK-9(H6) and MK-9(H8). The predominant cellular fatty acids were iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. The polar lipids profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and phosphatidylinositol mannoside, an unknown aminolipid and two unknown phospholipids. Phylogenetic analysis of 16S rRNA gene sequences showed the strain formed distinct clade within the genus Streptomyces and was closely related to Streptomyces echinatus NBRC 12763T (98.78 % 16S rRNA gene sequence similarity). According to the polyphasic approach as well as DNA-DNA relatedness, the strain could be clearly differentiated from closely related species and represents a novel species of the genus Streptomyces, for which the name Streptomyces actinomycinicus sp. nov. is proposed. The type strain is RCU-197T ( = JCM 30864T = TISTR 2208T = PCU 342T).

  19. Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut. [Streptomyces viridosporus

    SciTech Connect

    Pasti, M.B.; Crawford, D.L. ); Pometto, A.L., III ); Nuti, M.P. )

    1990-07-01

    The lignocellulose-degrading abilities of 11 novel actinomycete strains isolated from termite gut were determined and compared with that of the well-characterized actinomycete, Streptomyces viridosporus T7A. Lignocellulose bioconversion was followed by (i) monitoring the degradation of ({sup 14}C)lignin- and ({sup 14}C)cellulose-labeled phloem of Abies concolor to {sup 14}CO{sub 2} and {sup 14}C-labeled water-soluble products, (ii) determining lignocellulose, lignin, and carbohydrate losses resulting from growth on a lignocellulose substrate prepared from corn stalks (Zea mays), and (iii) quantifying production of a water-soluble lignin degradation intermediate (acid-precipitable polymeric lignin). Of the assays used, total lignocellulose weight loss was most useful in determining overall bioconversion ability but not in identifying the best lignin-solubilizing strains. A screening procedure based on {sup 14}CO{sub 2} evolution from ({sup 14}C-lignin)lignocellulose combined with measurement of acid-precipitable polymeric lignin yield was the most effective in identifying lignin-solubilizing strains. For the termite gut strains, the pH of the medium showed no increase after 3 weeks of growth on lignocellulose. This is markedly different from the pattern observed with S. viridosporus T7A, which raises the medium pH considerably. Production of extracellular peroxidases by the 11 strains and S. viridosporus T7A was followed for 5 days in liquid cultures. On the basis of an increase of specific peroxidase activity in the presence of lignocellulose in the medium, the actinomycetes could be placed into the same three groups.

  20. Streptomyces alfalfae sp. nov. and comparisons with its closest taxa Streptomyces silaceus, Streptomyces flavofungini and Streptomyces intermedius.

    PubMed

    She, Wenqing; Sun, Zhongfeng; Yi, Lei; Zhao, Shumiao; Liang, Yunxiang

    2016-01-01

    A novel streptomycete strain, designated XY25T, was isolated from the rhizosphere soil in an alfalfa field in Jingyang, Shanxi, China. The isolate showed optimal growth at 37 °C, and was capable of growing at pH 6-10 and in the presence of 0-6 % (w/v) NaCl. Mycelia of strain XY25T appeared spiral and developed into white spore chains with long-rod spores and a smooth surface. The 16S rRNA gene sequence of XY25T was determined and was found to be highly similar to those of species of the genus Streptomyces including Streptomyces silaceus DSM 41861T (99.11 % 16S rRNA gene sequence similarity), Streptomyces flavofungini DSM 40366T (98.49 %) and Streptomyces intermedius DSM 40372T (98.43 %), all of which were used for further characterization. Each of the four streptomycetes showed distinctive patterns of carbon usage and fatty acids composition. Analysis of cellular components of strain XY25T revealed ll-diaminopimelic acid as diagnostic diamino acid and xylose as the major sugar, whereas polar lipids were determined as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, an unknown phospholipid, two unknown phosphatidylinositol mannosides and several unknown lipids. Menaquinones were dominated by MK-9(H6) and MK-9(H8), and the main fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. DNA-DNA hybridization studies indicated that strain XY25T showed relatedness values of 35.2-40.42 % with the closest related species. Based on these results, strain XY25T represents a novel species of the genus Streptomyces, for which the name Streptomyces alfalfae sp. nov. is proposed. The type strain is XY25T ( = KCTC 39571T = CCTCC AA2015019T).

  1. Draft Genome Sequence of Streptomyces sp. F-3

    PubMed Central

    Sun, Xiaomeng; Meng, Jing; Liu, Shijia; Zhang, Huaiqiang

    2016-01-01

    Streptomyces sp. F-3 is a kind of thermophilic Streptomyces strain that can produce cellulolytic enzymes and diverse secondary metabolites. Here, we report the complete genome of this organism, whose genome length is 5,303,958 bp, containing 6,041 protein-coding genes, 69 tRNA operons, and three rRNA operons. PMID:27492002

  2. CHARACTERIZATION AND BIOCONTROL POTENT OF STREPTOMYCES SP. ISOLATED FROM THE RHIZOSPHERE OF ONONIS ANGUSTISSIMA LAM.

    PubMed

    Ghadbane, M; Belhadj, H; Medjekal, S; Harzallah, D

    2015-01-01

    A total of 40 actinomycetes isolated from rhizosphere soils of Ononis angustissima Lam. were in vitro tested for their antagonism against deferent pathogenic microorganisms by streak assay. Among the isolates, four (21, 2A26, 1B10 and 2C34) present a potent antagonism against both pathogenic bacteria and fungi, they were selected, identified by 16S rDNA sequence analysis and phenotypic properties, and tested for their antimicrobial activity as well as their biocontrol potential against Chickpea (Cicer arietinum L.) pathogenic fungus (Fusarium oxysporum). Cultural characteristic studies strongly suggested that these strains belong to the genus Streptomyces. The four Streptomyces sp., solubilize phosphate and produce extracellular fungal cell-wall degrading enzymes chitinase and protease, as well as a marked production of acid-β-indole acetic (AIA). The nucleotide sequence of the 16S rRNA gene of Streptomyces sp. strains 21, 2A26, 1B10 and 2C34 exhibited close similarity (62-75%) with Streptomyces parvulus MARS 16S rRNA genes. The inhibition was higher against fungi and Gram+ bacteria, while Gram- bacteria were less inhibited. The growth of the plant pathogenic fungus Fusarium oxysporum was considerably inhibited in the presence of the strains 21, 2A26, 1B10 and 2C34 culture supernatant. These studies revealed that the presence of the Streptomyces strains in the soil significantly promoted the growth of the Chickpea plants. These results indicate that the Streptomyces strains isolated for rhizosphere from Ononis angustissima Lam. growing in arid conditions in southern Algeria (Sahara) could be an interesting source for antimicrobial bioactive substances and as biocontrol agents.

  3. Saccharothrix hoggarensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-02-01

    An actinomycete, designated SA181(T), was isolated from Saharan soil in the Hoggar region (south Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the isolate were consistent with the genus Saccharothrix, and 16S rRNA gene sequence analysis confirmed that strain SA181(T) was a novel member of the genus Saccharothrix. DNA-DNA hybridization values between strain SA181(T) and its closest phylogenetic neighbours, the type strains of Saccharothrix longispora, Saccharothrix texasensis and Saccharothrix xinjiangensis, were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the isolate represents a novel species of the genus Saccharothrix, for which the name Saccharothrix hoggarensis sp. nov. is proposed, with the type strain SA181(T) ( = DSM 45457(T)  = CCUG 60214(T)).

  4. Actinoalloteichus hoggarensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boudjelal, Farida; Zitouni, Abdelghani; Bouras, Noureddine; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-06-01

    A moderately halophilic actinomycete strain, designated AH97T, was isolated from Saharan soil in the Hoggar region (south Algeria) and was subjected to polyphasic taxonomic characterization. The morphological and chemotaxonomic characteristics of the strain were consistent with those of the genus Actinoalloteichus. Results of 16S rRNA gene sequence comparison revealed that strain AH97T shared the highest degree of 16S rRNA gene sequence similarity with Actinoalloteichus hymeniacidonis DSM 45092T (99.3 %) and Actinoalloteichus nanshanensis DSM 45655T (98.7 %). However, DNA-DNA hybridization studies showed only 26.5 % relatedness with A. hymeniacidonis DSM 45092T and 28.0 % with A. nanshanensis DSM 45655T. The genotypic and phenotypic data showed that strain AH97T represents a novel species of the genus Actinoalloteichus, for which the name Actinoalloteichus hoggarensis sp. nov. is proposed, with AH97T ( = DSM 45943T = CECT 8639T) as the type strain.

  5. Saccharothrix tamanrassetensis sp. nov., an actinomycete isolated from Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2015-04-01

    Actinomycete strain SA198(T), isolated from a Saharan soil sample of Algeria, was characterized taxonomically by using a polyphasic approach. Chemotaxonomic and morphological characteristics observed suggested that it was a member of the genus Saccharothrix . The 16S rRNA gene sequence analysis confirmed that strain SA198(T) was a member of the genus Saccharothrix and showed a similarity level ranging between 97.5 and 98.9% within species of the genus Saccharothrix , Saccharothrix australiensis being the most closely related. However, DNA-DNA hybridization values between strain SA198(T) and its closest phylogenetic neighbours, the type strains of S. australiensis , Saccharothrix xinjiangensis , Saccharothrix algeriensis and Saccharothrix espanaensis , were clearly below the 70% threshold. Based upon genotypic and phenotypic differences from other members of the genus, a novel species, Saccharothrix tamanrassetensis sp. nov., is proposed, with SA198(T) ( = DSM 45947(T) = CECT 8640(T)) as the type strain.

  6. Salinispora pacifica sp. nov., an actinomycete from marine sediments.

    PubMed

    Ahmed, Lina; Jensen, Paul R; Freel, Kelle C; Brown, Ros; Jones, Amanda L; Kim, Byung-Yong; Goodfellow, Michael

    2013-05-01

    A polyphasic analysis was carried out to clarify the taxonomic status of four marine actinomycete strains that share a phylogenetic relationship and phenotypic characteristics with the genus Salinispora. These strains formed a distinct lineage within the Salinispora 16S rRNA and gyrB trees and were found to possess a range of phenotypic properties and DNA:DNA hybridization values that distinguished them from the type strains of the two validly named species in this genus, Salinispora tropica (CNB-440(T), ATCC BAA-916(T)) and Salinispora arenicola (CNH-643(T), ATCC BAA-917(T)). The combined genotypic and phenotypic data support this conclusion. It is proposed that the strains be designated as Salinispora pacifica sp. nov., the type strain of which is CNR-114(T) (DSMZ YYYYT = KACC 17160(T)).

  7. Terrabacter lapilli sp. nov., an actinomycete isolated from stone.

    PubMed

    Lee, Jeong-Eon; Seo, Jae Pyo; Lee, Dong Wan; Ko, Young-Hwan; Lee, Soon Dong

    2008-05-01

    A novel actinomycete, designated strain LR-26T, was isolated from a small stone collected from an agricultural field in Jeju, Republic of Korea. Cells of the organism were strictly aerobic, Gram-positive, non-motile, short rods. Colonies were bright yellow, circular, smooth and translucent. The organism was characterized chemotaxonomically as having ll-diaminopimelic acid in the cell wall, MK-8(H4) as major menaquinone, a polar lipid profile including diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and unknown phospholipids, iso-C15 : 0 as the predominant fatty acid and a DNA G+C content of 72.6 mol%. Comparative 16S rRNA gene sequence analysis showed that the organism was related to the genera Intrasporangium, Terracoccus and Terrabacter within the family Intrasporangiaceae. The closest phylogenetic relatives of strain LR-26T were the type strains of Terrabacter terrae (99.4 % 16S rRNA gene sequence similarity), Terrabacter aerolatus (99.3 %) and Terrabacter tumescens (99.3 %). DNA-DNA hybridization experiments showed that strain LR-26T shared low levels of DNA-DNA relatedness with Terrabacter terrae LMG 22921T (17.6 and 22.8 % from reciprocal experiments) and with Terrabacter tumescens IMSNU 21313T (27.6 and 34.9 %). The phenotypic data and low levels of DNA-DNA relatedness readily distinguished strain LR-26T from the type strains of recognized species of the genus Terrabacter, and showed that it therefore represents a novel species. The name Terrabacter lapilli sp. nov. is proposed for this novel actinomycete. The type strain is LR-26T (=JBRI 2002T =KCTC 19199T =DSM 18583T).

  8. Streptomyces mangrovi sp. nov., isolated from mangrove forest sediment.

    PubMed

    Yousif, Ghada; Busarakam, Kanungnid; Kim, Byung-Yong; Goodfellow, Michael

    2015-09-01

    A Streptomyces strain isolated from a mangrove sediment was classified using a polyphasic approach. The organism, isolate GY1(T), was found to have chemical and morphological properties typical of members of the genus Streptomyces. The isolate was shown to form a distinct phyletic line within the Streptomyces radiopugnans 16S rRNA gene subclade and to be closely related to the type strain of Streptomyces fenhuangensis (98.7 % similarity). It is also closely related to the type strain of Streptomyces bakulensis which was also closely related to members of the Streptomyces glaucosporus 16S rRNA gene subclade. Isolate GY1(T) was distinguished readily from the S. barkulensis type strain and from species classified in the S. radiopugnans clade using a combination of morphological and physiological properties, including a requirement for seawater for growth. Based on the genotypic and phenotypic data, it is proposed that isolate GY1(T) (=NCIMB 14980(T), NRRL B-69296(T)) be classified in the genus Streptomyces as Streptomyces mangrovi sp. nov.

  9. Microbispora bryophytorum sp. nov., an actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Zhang, Yuejing; Liu, Chongxi; Wang, Haiyan; Zhao, Junwei; Li, Lianjie; Zhang, Zhongwen; Wang, Xiangjing; Xiang, Wensheng

    2015-04-01

    A novel endophytic actinomycete, designated strain NEAU-TX2-2(T), was isolated from moss and characterized using a polyphasic approach. The isolate was found to have morphological characteristics typical of the genus Microbispora . The isolate formed longitudinally paired spores on the tips of short sporophores that branched from aerial hyphae. Analysis of the 16S rRNA gene sequence supported the assignment of the novel strain to the genus Microbispora , and strain NEAU-TX2-2(T) exhibited 99.08 and 98.62% gene sequence similarities to Microbispora amethystogenes JCM 3021(T) and Microbispora rosea subsp. rosea JCM 3006(T), respectively. However two tree-making algorithms supported the position that strain NEAU-TX2-2(T) formed a distinct clade with M. rosea subsp. rosea JCM 3006(T). A low level of DNA-DNA relatedness allowed the isolate to be differentiated from M. amethystogenes JCM 3021(T) and M. rosea subsp. rosea JCM 3006(T). Moreover, strain NEAU-TX2-2(T) could also be distinguished from its closest phylogenetic relatives by morphological and physiological characteristics. Therefore, it is proposed that strain NEAU-TX2-2(T) represents a novel species of the genus Microbispora for which the name Microbispora bryophytorum sp. nov. is proposed. The type strain is NEAU-TX2-2(T) ( = CGMCC 4.7138(T) = DSM 46710(T)).

  10. Streptomyces hypolithicus sp. nov., isolated from an Antarctic hypolith community.

    PubMed

    Le Roes-Hill, Marilize; Rohland, Jeffrey; Meyers, Paul R; Cowan, Don A; Burton, Stephanie G

    2009-08-01

    As part of an enzyme-screening programme, an actinobacterium, strain HSM#10T, was isolated from a sample collected from the base of a translucent quartz rock in Miers Valley, eastern Antarctica. The isolate produced branching vegetative mycelium that was characteristic of filamentous actinobacteria. The chemotaxonomic characteristics of the strain suggested that HSM#10T should be classified as a member of the genus Streptomyces. Furthermore, phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was closely related to members of the genus Streptomyces, which supports the classification of this strain within the family Streptomycetaceae. Phenotypic and phylogenetic results allowed strain HSM#10T to be differentiated from known streptomycetes. DNA-DNA hybridization data also showed that strain HSM#10T could be differentiated from its nearest phylogenetic neighbours Streptomyces chryseus DSM 40420T (53.55+/-3.15% DNA relatedness), Streptomyces helvaticus DSM 40431T (38.75+/-2.75%), Streptomyces flavidovirens DSM 40150T (30.7+/-2.90%) and Streptomyces albidochromogenes DSM 41800T (33.9+/-0.10%). Therefore, the name Streptomyces hypolithicus sp. nov. is proposed, with HSM#10T (=DSM 41950T=NRRL B-24669T) as the type strain.

  11. Hormaomycins B and C: New Antibiotic Cyclic Depsipeptides from a Marine Mudflat-Derived Streptomyces sp.

    PubMed Central

    Bae, Munhyung; Chung, Beomkoo; Oh, Ki-Bong; Shin, Jongheon; Oh, Dong-Chan

    2015-01-01

    Alterations in microbial culture conditions may trigger the production of diverse bioactive secondary metabolites. While applying various culture conditions and monitoring secondary metabolite profiles using LC/MS, hormaomycins B and C (1 and 2) were discovered from a marine mudflat-derived actinomycete, Streptomyces sp., collected in Mohang, Korea. The planar structures of the hormaomycins, which bear structurally-unique units, such as 4-(Z)-propenylproline, 3-(2-nitrocyclopropyl)alanine, 5-chloro-1-hydroxypyrrol-2-carboxylic acid and β-methylphenylalanine, were established as the first natural analogues belonging to the hormaomycin peptide class. The absolute configurations of 1 and 2 were deduced by comparing their CD spectra with that of hormaomycin. These hormaomycins exhibited significant inhibitory effects against various pathogenic Gram-positive and Gram-negative bacteria. PMID:26287218

  12. Novel propanamide analogue and antiproliferative diketopiperazines from mangrove Streptomyces sp. Q24.

    PubMed

    Ye, Xuewei; Chai, Weiyun; Lian, Xiao-Yuan; Zhang, Zhizhen

    2017-06-01

    A new propanamide analogue (1), along with one known alkaloid (2) and four known diketopiperazines (3-6), was isolated from a cultured broth of the actinomycete Streptomyces sp. Q24 that was obtained from a sample of mangrove soil. The structures of these isolates were characterised as 3-acetylamino-N-2-thienyl-propanamide (1), N-acetyltryptamine (2), cyclo-(l-phenylalanine-l-4-hydroxyproline) (3), cyclo-(l-leucine-l-4-hydroxyproline) (4), cyclo-(l-phenylalanine-d-4-hydroxyproline) (5) and cyclo-(l-leucine-l-proline) (6) based on their NMR and HRESIMS data as well as optical rotation. Three diketopiperazines (3, 4, 6) showed activity in inhibiting the proliferation of human glioma U87-MG and U251 cells. This type of the new propanamide analogue (1) is first found from a nature source and the antiproliferative property of these three diketopiperazines against glioma cells is also reported herein for the first time.

  13. Marine Sponge-Derived Streptomyces sp. SBT343 Extract Inhibits Staphylococcal Biofilm Formation

    PubMed Central

    Balasubramanian, Srikkanth; Othman, Eman M.; Kampik, Daniel; Stopper, Helga; Hentschel, Ute; Ziebuhr, Wilma; Oelschlaeger, Tobias A.; Abdelmohsen, Usama R.

    2017-01-01

    Staphylococcus epidermidis and Staphylococcus aureus are opportunistic pathogens that cause nosocomial and chronic biofilm-associated infections. Indwelling medical devices and contact lenses are ideal ecological niches for formation of staphylococcal biofilms. Bacteria within biofilms are known to display reduced susceptibilities to antimicrobials and are protected from the host immune system. High rates of acquired antibiotic resistances in staphylococci and other biofilm-forming bacteria further hamper treatment options and highlight the need for new anti-biofilm strategies. Here, we aimed to evaluate the potential of marine sponge-derived actinomycetes in inhibiting biofilm formation of several strains of S. epidermidis, S. aureus, and Pseudomonas aeruginosa. Results from in vitro biofilm-formation assays, as well as scanning electron and confocal microscopy, revealed that an organic extract derived from the marine sponge-associated bacterium Streptomyces sp. SBT343 significantly inhibited staphylococcal biofilm formation on polystyrene, glass and contact lens surfaces, without affecting bacterial growth. The extract also displayed similar antagonistic effects towards the biofilm formation of other S. epidermidis and S. aureus strains tested but had no inhibitory effects towards Pseudomonas biofilms. Interestingly the extract, at lower effective concentrations, did not exhibit cytotoxic effects on mouse fibroblast, macrophage and human corneal epithelial cell lines. Chemical analysis by High Resolution Fourier Transform Mass Spectrometry (HRMS) of the Streptomyces sp. SBT343 extract proportion revealed its chemical richness and complexity. Preliminary physico-chemical characterization of the extract highlighted the heat-stable and non-proteinaceous nature of the active component(s). The combined data suggest that the Streptomyces sp. SBT343 extract selectively inhibits staphylococcal biofilm formation without interfering with bacterial cell viability. Due to

  14. Marine Sponge-Derived Streptomyces sp. SBT343 Extract Inhibits Staphylococcal Biofilm Formation.

    PubMed

    Balasubramanian, Srikkanth; Othman, Eman M; Kampik, Daniel; Stopper, Helga; Hentschel, Ute; Ziebuhr, Wilma; Oelschlaeger, Tobias A; Abdelmohsen, Usama R

    2017-01-01

    Staphylococcus epidermidis and Staphylococcus aureus are opportunistic pathogens that cause nosocomial and chronic biofilm-associated infections. Indwelling medical devices and contact lenses are ideal ecological niches for formation of staphylococcal biofilms. Bacteria within biofilms are known to display reduced susceptibilities to antimicrobials and are protected from the host immune system. High rates of acquired antibiotic resistances in staphylococci and other biofilm-forming bacteria further hamper treatment options and highlight the need for new anti-biofilm strategies. Here, we aimed to evaluate the potential of marine sponge-derived actinomycetes in inhibiting biofilm formation of several strains of S. epidermidis, S. aureus, and Pseudomonas aeruginosa. Results from in vitro biofilm-formation assays, as well as scanning electron and confocal microscopy, revealed that an organic extract derived from the marine sponge-associated bacterium Streptomyces sp. SBT343 significantly inhibited staphylococcal biofilm formation on polystyrene, glass and contact lens surfaces, without affecting bacterial growth. The extract also displayed similar antagonistic effects towards the biofilm formation of other S. epidermidis and S. aureus strains tested but had no inhibitory effects towards Pseudomonas biofilms. Interestingly the extract, at lower effective concentrations, did not exhibit cytotoxic effects on mouse fibroblast, macrophage and human corneal epithelial cell lines. Chemical analysis by High Resolution Fourier Transform Mass Spectrometry (HRMS) of the Streptomyces sp. SBT343 extract proportion revealed its chemical richness and complexity. Preliminary physico-chemical characterization of the extract highlighted the heat-stable and non-proteinaceous nature of the active component(s). The combined data suggest that the Streptomyces sp. SBT343 extract selectively inhibits staphylococcal biofilm formation without interfering with bacterial cell viability. Due to

  15. Streptomyces lonarensis sp. nov., isolated from Lonar Lake, a meteorite salt water lake in India.

    PubMed

    Sharma, Trupti K; Mawlankar, Rahul; Sonalkar, Vidya V; Shinde, Vidhya K; Zhan, Jing; Li, Wen-Jun; Rele, Meenakshi V; Dastager, Syed G; Kumar, Lalitha Sunil

    2016-02-01

    A novel alkaliphilic actinomycete, strain NCL716(T), was isolated from a soil sample collected from the vicinity of Lonar Lake, an alkaline salt water meteorite lake in Buldhana district of Maharashtra State in India. The strain was characterised using a polyphasic taxonomic approach which confirmed that it belongs to the genus Streptomyces. Growth was observed over a pH range of 7-11 at 28 °C. The cell wall was found to contain LL-diaminopimelic acid and traces of meso-diaminopimelic acid. The major fatty acid components were identified as iso-C16:0 (46.8 %), C17:1 (12.4 %), anteiso-C15:0 (5.1 %) and anteiso-C17:1 (4.8 %). The major polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol. The major menaquinones were determined to be MK-9 (H6) (70.3 %), MK-9 (H4) (15.5 %) and MK-9 (H8) (7.2 %). The G+C content of the DNA of the type strain was determined to be 71.4 mol %. The 16S rRNA gene sequence has been deposited in GenBank with accession number FJ919811. Although the 16S rRNA gene sequence analysis revealed that strain NCL716(T) shares >99 % similarity with that of Streptomyces bohaiensis strain 11A07(T), DNA-DNA hybridization revealed only 33.2 ± 3.0 % relatedness between them. Moreover, these two strains can be readily distinguished by some distinct phenotypic characteristics. Hence, on the basis of phenotypic and genetic analyses, it is proposed that strain NCL716(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces lonarensis sp. nov., is proposed. The type strain is NCL 716(T) (=DSM 42084(T) = MTCC 11708(T) = KCTC 39684(T)).

  16. Streptomyces zhihengii sp. nov., isolated from rhizospheric soil of Psammosilene tunicoides.

    PubMed

    Huang, Mei-Juan; Fei, Jing-Jing; Salam, Nimaichand; Kim, Chang-Jin; Hozzein, Wael N; Xiao, Min; Huang, Hai-Quan; Li, Wen-Jun

    2016-10-01

    An actinomycete strain, designated YIM T102(T), was isolated from the rhizospheric soil of Psammosilene tunicoides W. C. Wu et C. Y. Wu collected from Lijiang, Yunnan Province, China. The taxonomic position of the new isolate was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM T102(T) belongs to the genus Streptomyces. Strain YIM T102(T) was most closely related to Streptomyces eurocidicus NRRL B-1676(T) with a pairwise 16S rRNA gene sequence similarity of 98.9 %. However, DNA-DNA relatedness value between strain YIM T102(T) and S. eurocidicus NBRC 13491(T) was found to be 37.8 ± 1.8 %. The menaquinone composition detected for strain YIM T102(T) was MK-9 (H6) and MK-9 (H8), while the major fatty acids were summed feature 4 (38.0 %), anteiso-C15:0 (13.1 %), iso-C16:0 (10.1 %), summed feature 3 (9.8 %) and C16:0 (9.0 %) and iso-C15:0 (5.2 %). The whole-cell hydrolysates contained galactose, glucose, ribose and mannose, along with LL-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The DNA G+C content was 70.7 mol%. Strain YIM T102(T) also exhibited antagonistic activity against Alternaria alternata, Alternaria brassicae and Colletotrichum nicotianae Averna, based on the findings from the comparative analyses of phenotypic and genotypic characteristics; it is proposed that strain YIM T102 represents a novel species of the genus Streptomyces, for which the name Streptomyces zhihengii sp. nov. is proposed. The type strain is YIM T102(T) (=KCTC 39115(T) = DSM 42176(T) = CGMCC 4.7248(T)).

  17. Exploring the Antimicrobial and Antitumor Potentials of Streptomyces sp. AGM12-1 Isolated from Egyptian Soil

    PubMed Central

    Ahmad, Maged S.; El-Gendy, Ahmed O.; Ahmed, Rasha R.; Hassan, Hossam M.; El-Kabbany, Hussein M.; Merdash, Ahmed G.

    2017-01-01

    The occurrence of extensive antibiotics resistant bacteria increased the demands for mining out new sources of antimicrobial agents. Actinomycetes, especially Streptomyces sp. have grasped considerable attention worldwide due to production of many useful bioactive metabolites. In the present study, a total of 52 actinomycetes were isolated from agricultural soil samples in Beni-Suef, Egypt. All isolates were characterized based on colony morphology, mycelium coloration, and pigment diffusion. They were screened for their capabilities to show antimicrobial activities against different indicator microorganisms, and only 20 isolates have shown significant antimicrobial activities against at least one of the tested indicator microorganisms. The isolate AGM12-1 was active against all tested microorganisms and showed a marked antitumor activity with IC50 3.3 and 1.1 μg/ml against HCT-116 and HepG-2 cell lines respectively. It was genotypically characterized as Streptomyces sp. with the presence of PKS Π biosynthetic gene cluster. Mannitol, ammonium sulfate, pH 7, 2% inoculum size and incubation for 11 days at 30°C were the optimum conditions that used to maximize the production and hence allowed purification of one active antimicrobial compound to homogeneity using high performance liquid chromatography with a molecular mass of m/z 488.05. Nuclear magnetic resonance structural elucidation showed that this compound was a diketopiperazine derivative. PMID:28348553

  18. Production and partial characterization of biosurfactant produced by Streptomyces sp. R1.

    PubMed

    Zambry, Nor Syafirah; Ayoib, Adilah; Md Noh, Nur Asshifa; Yahya, Ahmad Ramli Mohd

    2017-04-07

    The present study focused on developing a wild-type actinomycete isolate as a model for a non-pathogenic filamentous producer of biosurfactants. A total of 33 actinomycetes isolates were screened and their extracellular biosurfactants production was evaluated using olive oil as the main substrate. Out of 33 isolates, 32 showed positive results in the oil spreading technique (OST). All isolates showed good emulsification activity (E24) ranging from 84.1 to 95.8%. Based on OST and E24 values, isolate R1 was selected for further investigation in biosurfactant production in an agitated submerged fermentation. Phenotypic and genotypic analyses tentatively identified isolate R1 as a member of the Streptomyces genus. A submerged cultivation of Streptomyces sp. R1 was carried out in a 3-L stirred-tank bioreactor. The influence of impeller tip speed on volumetric oxygen transfer coefficient (k L a), growth, cell morphology and biosurfactant production was observed. It was found that the maximum biosurfactant production, indicated by the lowest surface tension measurement (40.5 ± 0.05 dynes/cm) was obtained at highest k L a value (50.94 h(-1)) regardless of agitation speed. The partially purified biosurfactant was obtained at a concentration of 7.19 g L(-1), characterized as a lipopeptide biosurfactant and was found to be stable over a wide range of temperature (20-121 °C), pH (2-12) and salinity [5-20% (w/v) of NaCl].

  19. New natural products identified by combined genomics-metabolomics profiling of marine Streptomyces sp. MP131-18

    PubMed Central

    Paulus, Constanze; Rebets, Yuriy; Tokovenko, Bogdan; Nadmid, Suvd; Terekhova, Larisa P.; Myronovskyi, Maksym; Zotchev, Sergey B.; Rückert, Christian; Braig, Simone; Zahler, Stefan; Kalinowski, Jörn; Luzhetskyy, Andriy

    2017-01-01

    Marine actinobacteria are drawing more and more attention as a promising source of new natural products. Here we report isolation, genome sequencing and metabolic profiling of new strain Streptomyces sp. MP131-18 isolated from marine sediment sample collected in the Trondheim Fjord, Norway. The 16S rRNA and multilocus phylogenetic analysis showed that MP131-18 belongs to the genus Streptomyces. The genome of MP131-18 isolate was sequenced, and 36 gene clusters involved in the biosynthesis of 18 different types of secondary metabolites were predicted using antiSMASH analysis. The combined genomics-metabolics profiling of the strain led to the identification of several new biologically active compounds. As a result, the family of bisindole pyrroles spiroindimicins was extended with two new members, spiroindimicins E and F. Furthermore, prediction of the biosynthetic pathway for unusual α-pyrone lagunapyrone isolated from MP131-18 resulted in foresight and identification of two new compounds of this family – lagunapyrones D and E. The diversity of identified and predicted compounds from Streptomyces sp. MP131-18 demonstrates that marine-derived actinomycetes are not only a promising source of new natural products, but also represent a valuable pool of genes for combinatorial biosynthesis of secondary metabolites. PMID:28186197

  20. New natural products identified by combined genomics-metabolomics profiling of marine Streptomyces sp. MP131-18.

    PubMed

    Paulus, Constanze; Rebets, Yuriy; Tokovenko, Bogdan; Nadmid, Suvd; Terekhova, Larisa P; Myronovskyi, Maksym; Zotchev, Sergey B; Rückert, Christian; Braig, Simone; Zahler, Stefan; Kalinowski, Jörn; Luzhetskyy, Andriy

    2017-02-10

    Marine actinobacteria are drawing more and more attention as a promising source of new natural products. Here we report isolation, genome sequencing and metabolic profiling of new strain Streptomyces sp. MP131-18 isolated from marine sediment sample collected in the Trondheim Fjord, Norway. The 16S rRNA and multilocus phylogenetic analysis showed that MP131-18 belongs to the genus Streptomyces. The genome of MP131-18 isolate was sequenced, and 36 gene clusters involved in the biosynthesis of 18 different types of secondary metabolites were predicted using antiSMASH analysis. The combined genomics-metabolics profiling of the strain led to the identification of several new biologically active compounds. As a result, the family of bisindole pyrroles spiroindimicins was extended with two new members, spiroindimicins E and F. Furthermore, prediction of the biosynthetic pathway for unusual α-pyrone lagunapyrone isolated from MP131-18 resulted in foresight and identification of two new compounds of this family - lagunapyrones D and E. The diversity of identified and predicted compounds from Streptomyces sp. MP131-18 demonstrates that marine-derived actinomycetes are not only a promising source of new natural products, but also represent a valuable pool of genes for combinatorial biosynthesis of secondary metabolites.

  1. Isolation, identification and screening of antimicrobial thermophilic Streptomyces sp. Al-Dhabi-1 isolated from Tharban hot spring, Saudi Arabia.

    PubMed

    Al-Dhabi, Naif Abdullah; Esmail, Galal Ali; Duraipandiyan, Veeramuthu; Valan Arasu, Mariadhas; Salem-Bekhit, Mounir M

    2016-01-01

    The strain Streptomyces sp. Al-Dhabi-1 was isolated from soil sediments collected from Tharban hot spring in the southern west of Saudi Arabia using actinomycetes isolation agar and starch casein agar at 55 °C. Identification of the isolate was done according to morphological, physiological and biochemical characteristics and 16S rRNA sequence similarity as well. 16S rRNA sequence and blast analyses confirmed that the isolate belonging to the genus Streptomyces. The sequence was submitted to GenBank with accession number (KF815080). Ethyl acetate extract of Streptomyces sp. Al-Dhabi-1 showed good antimicrobial activities against tested pathogenic microbes. Minimum inhibitory concentration results showed that the best values were observed against S. agalactiae (<0.039 mg/ml) and Klebsiella pneumonia (0.125 mg/ml). Minimum inhibitory concentration of Al-Dhabi-1 against fungi; Cryptococcus neoformans (0.078 mg/ml), C. albicans (0.156 mg/ml), A. niger (0.625 mg/ml), and T. mentagrophytes (0.156 mg/ml). GC-MS analysis was used for the chemical profile of ethyl acetate extract. Benzeneacetic acid (16.02 %) and acetic acid 2-phenylethyl ester (10.35 %) were the major compounds among 31 substances found the ethyl acetate extract. According to the results of antimicrobial activity against pathogenic microbes, it is clear that the actinomycetes from hot springs with extreme environments are promising source for antimicrobial compounds.

  2. Langkocyclines: novel angucycline antibiotics from Streptomyces sp. Acta 3034(*).

    PubMed

    Kalyon, Bahar; Tan, Geok-Yuan A; Pinto, John M; Foo, Cheau-Yee; Wiese, Jutta; Imhoff, Johannes F; Süssmuth, Roderich D; Sabaratnam, Vikineswary; Fiedler, Hans-Peter

    2013-10-01

    Langkocyclines A1-A3 and B1 and B2, five new angucycline antibiotics produced by Streptomyces sp. Acta 3034, were detected in the course of our HPLC-diode array screening. The producing strain was isolated from the rhizospheric soil of a Clitorea sp. collected from Burau Bay, Langkawi, Malaysia, and was characterized by morphological, physiological and chemotaxonomic features in addition to 16S ribosomal RNA gene sequence information. Strain Acta 3034 is closely related to Streptomyces psammoticus NBRC 13971(T) and Streptomyces lanatus NBRC 12787(T). Langkocyclines consist of an angular tetracyclic benz[a]anthracene skeleton and hydrolyzable O-glycosidic sugar moieties. The yellow-colored A-type langkocyclines differ in their aglycon from the blue-lilac-colored B-type langkocyclines. The A-type langkocycline aglycon is identical to that of aquayamycin and urdamycin A. The chemical structures of the langkocyclines were elucidated by HR-MS, 1D and 2D NMR experiments. They are biologically active against Gram-positive bacteria and exhibit a moderate antiproliferative activity against various human tumor cell lines.

  3. Antagonistic Effect of Streptomyces sp. BS062 against Botrytis Diseases

    PubMed Central

    Kim, Young-Sook; Lee, In-Kyoung

    2015-01-01

    The use of microorganisms and their secreted molecules to prevent plant diseases is considered an attractive alternative and way to supplement synthetic fungicides for the management of plant diseases. Strain BS062 was selected based on its ability to inhibit the mycelial growth of Botrytis cinerea, a major causal fungus of postharvest root rot of ginseng and strawberry gray mold disease. Strain BS062 was found to be closely related to Streptomyces hygroscopicus (99% similarity) on the basis of 16S ribosomal DNA sequence analysis. Postharvest root rot of ginseng and strawberry gray mold disease caused by B. cinerea were controlled up to 73.9% and 58%, respectively, upon treatment with culture broth of Streptomyces sp. BS062. These results suggest that strain BS062 may be a potential agent for controlling ginseng postharvest root rot and strawberry gray mold disease. PMID:26539052

  4. Biological Characteristics and Antimicrobial Activity of Endophytic Streptomyces sp. TQR12-4 Isolated from Elite Citrus nobilis Cultivar Ham Yen of Vietnam

    PubMed Central

    Mai-Linh, Nguyen Vu; Hong-Lien, Nguyen Thi; Van Hieu, Nguyen

    2016-01-01

    Ham Yen orange (Citrus nobilis Lour) is the highly valuable commercial fruit of Vietnam. With the blooming of fruit production and farming area, this specialty crop is facing threats from several serious diseases; therefore the search for new effective biocontrollers is required to prevent the existing excessive use of fertilizers and plant protection chemicals. Endophytic actinomycetes are of great scientific interest due to their high potential of application in agriculture and pharmaceutical research. In this work, endophytic actinomycetes were isolated from a native orange species of Northeast mountainous province Tuyen Quang. Among 49 isolates obtained, the isolate TQR12-4 strongly inhibited test pathogens Colletotrichum truncatum, Geotrichum candidum, Fusarium oxysporum, and F. udum. This isolate gave comparatively high biomass yields on different substrates, for example, carboxy methyl cellulose, starch, protein, and chitin, within a wide range of temperature from 15 to 45°C and pH from 4 to 10. Sequence analysis of 16S rDNA gene showed that TQR12-4 shared 99% similarity to Streptomyces prasinopilosus; however, it slightly differed from the latter in spore morphology and hence was named as Streptomyces sp. TQR12-4. A thermostable antifungal substance of nonpeptide nature produced by Streptomyces sp. TQR12-4 had MIC against Fusarium udum of 100 μg/mL and 400 μg/mL respective to extract fractions X4 and X5. PMID:27795709

  5. Draft Genome Sequence of the Streptothricin-Producing Strain Streptomyces sp. fd2-tb

    PubMed Central

    Tang, Biao; Yu, Yucong; Cen, Xufeng; Zhu, Yongqiang; Dai, Ruixue; Wang, Xianwei

    2015-01-01

    Streptomyces sp. fd2-tb can produce streptothricin class antibiotics with broad antimicrobial spectra. To better understand the mechanism of streptothricin biosynthesis and to assess the capacity of this strain in secondary metabolism, we report the draft genome sequence of Streptomyces sp. strain fd2-tb. PMID:26514767

  6. Thaxtomin A-deficient endophytic Streptomyces sp. enhances plant disease resistance to pathogenic Streptomyces scabies.

    PubMed

    Lin, Lan; Ge, Hui Ming; Yan, Tong; Qin, Yan Hua; Tan, Ren Xiang

    2012-12-01

    Each plant species in nature harbors endophytes, a community of microbes living within host plants without causing any disease symptom. However, the exploitation of endophyte-based phytoprotectants is hampered by the paucity of mechanistic understandings of endophyte-plant interaction. We here reported two endophytic Streptomyces isolates IFB-A02 and IFB-A03 recovered from a stress-tolerant dicotyledonous plant Artemisia annua L. After the determination of their non-pathogenicity at the genomic level and from the toxin (thaxtomin A, TXT) level, the endophytism of both isolates was supported by their successful colonization in planta. Of the two endophytes, IFB-A03 was further studied for the mechanism of endophyte-conferred phytoprotection owing to its plant growth promotion in model eudicot Arabidopsis thaliana. Using the endophyte-Arabidopsis co-cultivation system into which pathogenic Streptomyces scabies was introduced, we demonstrated that IFB-A03 pre-inoculation could activate the salicylic acid (SA)-mediated plant defense responses upon pathogen challenge. Moreover, IFB-A03 was shown to partially rescue the defense deficiency in eds5 (enhanced disease susceptibility 5) Arabidopsis mutants, putatively acting at the upstream of SA accumulation in the defense signaling pathway associated with the systemic acquired resistance (SAR). These data suggest that endophytic Streptomyces sp. IFB-A03 could be a promising candidate for biocontrol agents against S. scabies--a causative pathogen of common scab diseases prevailing in agronomic systems.

  7. Antagonistic activity of endo-β-1,3-glucanase from a novel isolate, Streptomyces sp. 9X166, against black rot in orchids.

    PubMed

    Sakdapetsiri, Chatsuda; Fukuta, Yasuhisa; Aramsirirujiwet, Yaovapa; Shirasaka, Norifumi; Kitpreechavanich, Vichien

    2016-05-01

    A total of 123 actinomycetes was isolated from 12 varieties of wild orchids and screened for potential antagonistic activity against Phytophthora, which causes black rot disease in orchids. In vitro and in vivo experimental results revealed that Streptomyces sp. strain 9X166 showed the highest antagonistic activity; its β-1,3-glucanase production ability was a key mechanism for growth inhibition of the pathogen. PCR amplification and DNA sequencing of the 16S ribosomal RNA gene allowed the identification of this strain, with high similarity (99.93%) to the novel species Streptomyces similaensis. The glucanase enzyme, purified to homogeneity by anion exchange and gel filtration chromatography, showed a specific activity of 58 U mg(-1) (a 3.9-fold increase) and yield of 6.4%. The molecular weight, as determined by SDS-PAGE and gel filtration, was approximately 99 and 80 kDa, respectively, suggesting that the enzyme was a monomer. The purified enzyme showed the highest substrate specificity to laminarin, indicating that it was β-1,3-glucanase. The hydrolyzed products of cello-oligosaccharides suggested that this enzyme was endo-type β-1,3-glucanase. Streptomyces sp. 9X166 culture filtrate, possessing β-1,3-glucanase activity, could degrade both freeze-dried and living mycelium. This is the first report on a β-1,3-glucanase-producing Streptomyces sp. that could be an effective biocontrol agent for black rot disease in orchids.

  8. Streptomyces leeuwenhoekii sp. nov., the producer of chaxalactins and chaxamycins, forms a distinct branch in Streptomyces gene trees.

    PubMed

    Busarakam, Kanungnid; Bull, Alan T; Girard, Geneviève; Labeda, David P; van Wezel, Gilles P; Goodfellow, Michael

    2014-05-01

    A polyphasic study was carried out to establish the taxonomic status of an Atacama Desert isolate, Streptomyces strain C34(T), which synthesises novel antibiotics, the chaxalactins and chaxamycins. The organism was shown to have chemotaxonomic, cultural and morphological properties consistent with its classification in the genus Streptomyces. Analysis of 16S rRNA gene sequences showed that strain C34(T) formed a distinct phyletic line in the Streptomyces gene tree that was very loosely associated with the type strains of several Streptomyces species. Multilocus sequence analysis based on five house-keeping gene alleles underpinned the separation of strain C34(T) from all of its nearest phylogenetic neighbours, apart from Streptomyces chiangmaiensis TA-1(T) and Streptomyces hyderabadensis OU-40(T) which are not currently in the MLSA database. Strain C34(T) was distinguished readily from the S. chiangmaiensis and S. hyderabadensis strains by using a combination of cultural and phenotypic data. Consequently, strain C34(T) is considered to represent a new species of the genus Streptomyces for which the name Streptomyces leeuwenhoekii sp. nov. is proposed. The type strain is C34(T) (= DSM 42122(T) = NRRL B-24963(T)). Analysis of the whole-genome sequence of S. leeuwenhoekii, with 6,780 predicted open reading frames and a total genome size of around 7.86 Mb, revealed a high potential for natural product biosynthesis.

  9. Draft genome sequences of three chemically rich actinomycetes isolated from Mediterranean sponges.

    PubMed

    Horn, Hannes; Cheng, Cheng; Edrada-Ebel, RuAngelie; Hentschel, Ute; Abdelmohsen, Usama Ramadan

    2015-12-01

    Metabolomic analysis has shown the chemical richness of the sponge-associated actinomycetes Streptomyces sp. SBT349, Nonomureae sp. SBT364, and Nocardiopsis sp. SBT366. The genomes of these actinomycetes were sequenced and the genomic potential for secondary metabolism was evaluated. Their draft genomes have sizes of 8.0, 10, and 5.8 Mb having 687, 367, and 179 contigs with a GC content of 71.6, 70.7, and 72.7%, respectively. Moreover, antiSMASH 3.0 predicted 108, 149, and 75 secondary metabolite gene clusters, respectively which highlight the metabolic capacity of the three actinomycete species to produce diverse classes of natural products.

  10. Violapyrones H and I, New Cytotoxic Compounds Isolated from Streptomyces sp. Associated with the Marine Starfish Acanthaster planci

    PubMed Central

    Shin, Hee Jae; Lee, Hwa-Sun; Lee, Jong Seok; Shin, Junho; Lee, Min Ah; Lee, Hyi-Seung; Lee, Yeon-Ju; Yun, Jieun; Kang, Jong Soon

    2014-01-01

    Two new α-pyrone derivatives, violapyrones H (1) and I (2), along with known violapyrones B (3) and C (4) were isolated from the fermentation broth of a marine actinomycete Streptomyces sp. The strain was derived from a crown-of-thorns starfish, Acanthaster planci, collected from Chuuk, Federated States of Micronesia. The structures of violapyrones were elucidated by the analysis of 1D and 2D NMR and HR-ESIMS data. Violapyrones (1–4) exhibited cytotoxicity against 10 human cancer cell lines with GI50 values of 1.10–26.12 μg/mL when tested using sulforhodamine B (SRB) assay. This is the first report on the cytotoxicity of violapyrones against cancer cell lines and the absolute configuration of violapyrone C. PMID:24886866

  11. Isolation, screening and partial purification of antimicrobial antibiotics from soil Streptomyces sp. SCA 7.

    PubMed

    Saravana Kumar, P; Duraipandiyan, V; Ignacimuthu, S

    2014-09-01

    Thirty-seven actinomycetes strains were isolated from soil samples collected from an agriculture field in Vengodu, Thiruvannamalai District, Tamil Nadu, India (latitude: 12° 54' 0033″, North; longitude: 79° 78' 5216″, East; elevation: 228.6/70.0 ft/m). The isolates were assessed for antagonistic activity against five Gram-positive bacteria, seven Gram-negative bacteria, and two pathogenic fungi. During the initial screening, 43% of the strains showed weak activity, 16% showed moderate activity, 5% showed good activity, and 35% showed no antagonistic activity. Among the strains tested, SCA 7 showed strong antimicrobial activity. Maximum biological activity was obtained on modified nutrient glucose agar (MNGA) medium. The mycelia of SCA 7 were extracted with methanol and tested against microbial pathogens using the disc diffusion method. The crude extract was purified partially using column chromatography and assessed for antimicrobial activity. Fraction 10 showed good activity against Staphylococcus epidermidis (31.25 μg/mL) and Malassezia pachydermatis (500 μg/mL) and the active principle (fraction 10) was identified as 2,4-bis (1,1-dimethylethyl) phenol. Based on morphological, physiological, biochemical, cultural, and molecular characteristics (16S rDNA sequencing), this strain was identified as Streptomyces sp. SCA 7. It could be used in the development of new substances for pharmaceutical or agricultural purposes.

  12. Phylum-specific regulation of resistomycin production in a Streptomyces sp. via microbial coculture.

    PubMed

    Carlson, Skylar; Tanouye, Urszula; Omarsdottir, Sesselja; Murphy, Brian T

    2015-03-27

    Actinomycete genomes are encoded with immense potential to produce secondary metabolites, however standard laboratory culture experiments rarely provide the conditions under which associated biosynthetic pathways are expressed. Despite years of research attempting to access these pathways and aside from a few well-studied bacterial quorum sensing systems, little is known about the specificity of secondary metabolite regulation in bacteria, such as the conditions under which a bacterium produces an antibiotic and the extent to which it does so in recognition of a particular species in the immediate environment. In the current study, we observed that the cocultivation of a Streptomyces sp. (strain B033) with four pathogenic strains of the phylum Proteobacteria resulted in the production of the antibiotic resistomycin. After further coculture experiments, we determined that Proteobacteria induced the production of resistomycin in B033 at significantly higher rates (65%) than strains from the phyla Firmicutes (5.9%) and Actinobacteria (9.1%), supporting that the regulation of secondary metabolism in bacteria can be dependent on the species present in the immediate environment. These results suggest a lack of promiscuity of antibiotic biosynthetic pathway regulation and indicate that it is feasible to mine existing microbial strain libraries for antibiotics in a phylum-specific manner.

  13. Streptomyces sp. TEM 33 possesses high lipolytic activity in solid-state fermentation in comparison with submerged fermentation.

    PubMed

    Cadirci, Bilge Hilal; Yasa, Ihsan; Kocyigit, Ali

    2016-01-01

    Solid-state fermentation (SSF) is a bioprocess that doesn't need an excess of free water, and it offers potential benefits for microbial cultivation for bioprocesses and product development. In comparing the antibiotic production, few detailed reports could be found with lipolytic enzyme production by Streptomycetes in SSF. Taking this knowledge into consideration, we prefer to purify Actinomycetes species as a new source for lipase production. The lipase-producing strain Streptomyces sp. TEM 33 was isolated from soil and lipase production was managed by solid-state fermentation (SSF) in comparison with submerged fermentation (SmF). Bioprocess-affecting factors like initial moisture content, incubation time, and various carbon and nitrogen additives and the other enzymes secreted into the media were optimized. Lipase activity was measured as 1.74 ± 0.0005 U/g dry substrate (gds) by the p-nitrophenylpalmitate (pNPP) method on day 6 of fermentation with 71.43% final substrate moisture content. In order to understand the metabolic priority in SSF, cellulase and xylanase activity of Streptomyces sp. TEM33 was also measured. The microorganism degrades the wheat bran to its usable form by excreting cellulases and xylanases; then it secretes the lipase that is necessary for degrading the oil in the medium.

  14. Exploitation of biological wastes for the production of value-added hydrolases by Streptomyces sp. MSWC1 isolated from municipal solid waste compost.

    PubMed

    Mokni-Tlili, Sonia; Ben Abdelmalek, Imen; Jedidi, Naceur; Belghith, Hafedh; Gargouri, Ali; Abdennaceur, Hassen; Marzouki, Mohamed Nejib

    2010-09-01

    Actinomycetes with the ability to degrade natural polysaccharides were isolated during a screening programme from soil, farmyard manure and municipal solid waste compost. One of the most potent isolates was identified as Streptomyces sp. MSWC1 using morphological and biochemical properties along with 16S rDNA partial sequence analysis. The highest enzyme production by Streptomyces was observed for the xylanase and chitinase activity on different carbon sources with an optimum of 12,100 IU ml(-1) and 110 IU ml(-1) at 3 days' culture on 1% of xylan and chitin, respectively. To meet the demand of industry, low-cost medium is required for the production of hydrolases by Streptomyces sp. Strain MSWC1 grown on manure, compost, and a natural carbon source was used to evaluate the re-utilisation of biological wastes for the production of value-added products. Despite the presence of a high amount of toxic heavy metals in the compost, Streptomyces produced interesting enzymes that have been biochemically characterized.

  15. Sannastatin, a novel toxic macrolactam polyketide glycoside produced by actinomycete Streptomyces sannanensis.

    PubMed

    Yang, Sheng-Xiang; Gao, Jin-Ming; Zhang, An-Ling; Laatsch, Hartmut

    2011-07-01

    A new rare 20-membered macrocyclic lactam incorporating a diene conjugated olefin, designated sannastatin (1), together with the known structurally related vicenistatin (2), has been isolated from the cultures of Streptomyces sannanensis, a bacteria found in the feces of Ailuropoda melanoleuca. The structure of the new compound was established on the basis of extensive spectroscopic analyses including 1D- and 2D-NMR ((1)H-(1)H COSY, TOCSY, HSQC, HMBC, and NOESY) experiments. Compounds 1 and 2 displayed significant growth inhibitory activity against the brine shrimp (Artemia salina) larvae.

  16. Actinomadura xylanilytica sp. nov., an actinomycete isolated from soil.

    PubMed

    Zucchi, Tiago Domingues; Kim, Byung-Yong; Bonda, Avinash Naga Venkata; Goodfellow, Michael

    2013-02-01

    The taxonomic position of a soil isolate, strain BK147(T), was established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Actinomadura. It formed a distinct phyletic line in the phylogenetic tree based on 16S rRNA gene sequences of members of the genus Actinomadura and was most closely, albeit loosely, related to Actinomadura bangladeshensis DSM 45347(T), Actinomadura meyerae DSM 44715(T) and Actinomadura napierensis NRRL B-24319(T) but was readily distinguished from these strains using a range of phenotypic properties. Based on the combined genotypic and phenotypic data it is proposed that isolate BK147(T) ( = KACC 20919(T) = NCIMB 14771(T) = NRRL B-24852(T)) be classified as the type strain of a novel species of the genus Actinomadura, for which the name Actinomadura xylanilytica sp. nov. is proposed.

  17. Study on bioactive compounds from Streptomyces sp. ANU 6277.

    PubMed

    Narayana, Kolla J P; Prabhakar, Peddikotla; Vijayalakshmi, Muvva; Venkateswarlu, Yenamandra; Krishna, Palakodety S J

    2008-01-01

    An attempt was made to study the bioactive compounds from a terrestrial Streptomyces sp. ANU 6277 isolated from laterite soil. Four active fractions were recovered from the solvent extracts obtained from the culture broth of five day-old strain. Three bioactive compounds were purified and identified as 3-phenylpropionic acid, anthracene-9,10-quinone and 8-hydroxyquinoline. The components of the partially purified fourth active fraction were analyzed by gas chromatography-mass spectrometry and identified as benzyl alcohol, phenylethyl alcohol and 2H-1, 4-benzoxazin-3 (4H)-one. Four active fractions were screened for antimicrobial activity against Gram-positive and Gram-negative bacteria, and fungi including phytopathogenic, toxigenic and dermatophytic genera. Among these metabolites, 8-hydroxyquinoline exhibited strong antibacterial and antifungal activity as compared to 3-phenylpropionic acid and anthracene-9,10-quinone.

  18. Bioactive Polycyclic Quinones from Marine Streptomyces sp. 182SMLY

    PubMed Central

    Liang, Ying; Xie, Xin; Chen, Lu; Yan, Shilun; Ye, Xuewei; Anjum, Komal; Huang, Haocai; Lian, Xiaoyuan; Zhang, Zhizhen

    2016-01-01

    Chemical investigation of the cultures of marine Streptomyces sp. 182SMLY led to the discovery of two new polycyclic anthraquinones, which were elucidated as N-acetyl-N-demethylmayamycin (1) and streptoanthraquinone A (2) based on the extensive spectroscopic analysis including 2D NMR, HRESIMS, and an electronic circular dichroism (ECD) calculation. Both anthraquinones remarkably suppressed the proliferation of four different glioma cell lines with IC50 values in a range from 0.5 to 7.3 μM and induced apoptosis in the glioma cells. The ratios of IC50 for normal human astrocytes to IC50 for glioma cells were 6.4–53 for 1 and >14–31 for 2. N-acetyl-N-demethylmayamycin (1) also inhibited the growth of methicillin-resistant Staphylococcus aureus with MIC 20.0 μM. PMID:26751456

  19. A novel PHB depolymerase from a thermophilic Streptomyces sp.

    PubMed

    Calabia, Buenaventurada P; Tokiwa, Yutaka

    2006-03-01

    A novel PHB depolymerase from a thermophilic Streptomyces sp. MG was purified to homogeneity by hydrophobic interaction chromatography and gel filtration. The molecular mass of the purified enzyme was 43 kDa as determined by size exclusion chromatography and 41 kDa by SDS-PAGE. The optimum pH and temperature were 8.5 and 60 degrees C respectively. The enzyme was stable at 50 degrees C and from pH 6.5-8.5. The enzyme hydrolyzed not only bacterial polyesters, i.e. poly(3-hydroxybutyric acid and poly(3-hydroxybutyrate-co-3-hydroxyvalerate), but also synthetic, aliphatic polyesters such as polypropiolactone, poly(ethylene adipate) and poly(ethylene succinate).

  20. A New Benzofuran Glycoside and Indole Alkaloids from a Sponge-Associated Rare Actinomycete, Amycolatopsis sp.

    PubMed Central

    Kwon, Yun; Kim, Seong-Hwan; Shin, Yoonho; Bae, Munhyung; Kim, Byung-Yong; Lee, Sang Kook; Oh, Ki-Bong; Shin, Jongheon; Oh, Dong-Chan

    2014-01-01

    Three new secondary metabolites, amycofuran (1), amycocyclopiazonic acid (2), and amycolactam (3), were isolated from the sponge-associated rare actinomycete Amycolatopsis sp. Based on combined spectroscopic analyses, the structures of 1–3 were determined to be a new benzofuran glycoside and new indole alkaloids related to cyclopiazonic acids, a class that has previously only been reported in fungi. The absolute configurations of 1 and 3 were deduced by ECD calculations, whereas that of 2 was determined using the modified Mosher method. Amycolactam (3) displayed significant cytotoxicity against the gastric cancer cell line SNU638 and the colon cancer cell line HCT116. PMID:24759001

  1. Rhodococcus kyotonensis sp. nov., a novel actinomycete isolated from soil.

    PubMed

    Li, Bing; Furihata, Keiko; Ding, Lin-Xian; Yokota, Akira

    2007-09-01

    A polyphasic study was undertaken to establish the taxonomic position of an isolate, strain DS472(T), from soil in Kyoto, Japan. Phylogenetic analysis, based on the 16S rRNA gene sequences, revealed that this strain constitutes a new subline within the genus Rhodococcus, with Rhodococcus yunnanensis YIM 70056(T) and Rhodococcus fascians DSM 20669(T) as its nearest phylogenetic neighbours (98.2 and 97.8 % sequence similarity, respectively). DNA-DNA hybridization experiments revealed 36 and 29 % relatedness between the isolate and its phylogenetic relatives, R. yunnanensis and R. fascians, respectively. Chemotaxonomic characteristics, including the major quinone MK-8(H(2)), predominant fatty acids C(16 : 0), C(18 : 1)omega9c and 10-methyl C(18 : 0), the presence of cell-wall chemotype IV and mycolic acids, were consistent with the properties of members of the genus Rhodococcus. The DNA G+C content was 64.5 mol%. On the basis of both phenotypic and genotypic evidence, strain DS472(T) represents a novel species of the genus Rhodococcus, for which the name Rhodococcus kyotonensis sp. nov. is proposed. The type strain is strain DS472(T) (=IAM 15415(T)=CCTCC AB206088(T)).

  2. Streptomyces fractus sp. nov., a novel streptomycete isolated from the gut of a South African termite.

    PubMed

    Rohland, Jeffrey; Meyers, Paul R

    2015-05-01

    An actinobacterial strain, MV32(T), was isolated from the paunch region of the hindgut of a South African termite, Amitermes hastatus, as part of an investigation of the actinobacterial population residing within this higher order termite species. Strain MV32(T) was chosen for further study from amongst the many potentially novel actinomycete isolates because of its strong antibacterial activity against Mycobacterium aurum A+. 16S rRNA gene phylogenetic analyses clearly placed strain MV32(T) within the genus Streptomyces, with 99.3% sequence similarity to its closest relative, Streptomyces endophyticus YIM 65594(T). Despite this high sequence similarity, DNA-DNA hybridisation analysis showed a DNA relatedness value of 62 ± 2%, to S. endophyticus DSM 41984(T) (indicating that strain MV32(T) belongs to a different genomic species), as well as values of 14.4 ± 0.8 and 10.4 ± 2.9%, respectively, to its next closest relatives, Streptomyces kunmingensis NRRL B-16240(T) and Streptomyces cinnabarinus NRRL B-12382(T). Based on these results and supported by both chemotaxonomic data and a number of phenotypic differences, strain MV32(T) is proposed to represent a new species within the genus Streptomyces, with the name Streptomyces fractus (= DSM 42163(T) = NRRL B-59159(T)).

  3. Biodegradation of anthracene by a novel actinomycete, Microbacterium sp. isolated from tropical hydrocarbon-contaminated soil.

    PubMed

    Salam, Lateef B; Obayori, Oluwafemi S; Olatoye, Nojeem O

    2014-01-01

    A novel anthracene-degrading Gram-positive actinomycete, Microbacterium sp. strain SL10 was isolated from a hydrocarbon-contaminated soil at a mechanical engineering workshop in Lagos, Nigeria. The polluted soil had an unusually high total hydrocarbon content of 157 g/kg and presence of various heavy metals. The isolate tolerated salt concentration of more than 4%. It resisted cefotaxime, streptomycin and ciprofloxacin, but susceptible to meropenem, linezolid and vancomycin. The isolate exhibited growth rate and doubling time of 0.82 days(-1) and 0.84 days, respectively on anthracene. It degraded 57.5 and 90.12% of anthracene within 12 and 21 days, respectively while the rate of anthracene utilization by the isolate was 4.79 mg l(-1) d(-1). To the best of our knowledge, this is the first report of isolation and characterization of anthracene-degrading Microbacterium sp.

  4. Cloning of the staurosporine biosynthetic gene cluster from Streptomyces sp. TP-A0274 and its heterologous expression in Streptomyces lividans.

    PubMed

    Onaka, Hiroyasu; Taniguchi, Shin-ichi; Igarashi, Yasuhiro; Furumai, Tamotsu

    2002-12-01

    Staurosporine is a representative member of indolocarbazole antibiotics. The entire staurosporine biosynthetic and regulatory gene cluster spanning 20-kb was cloned from Streptomyces sp. TP-A0274 and sequenced. The gene cluster consists of 14 ORFs and the amino acid sequence homology search revealed that it contains three genes, staO, staD, and staP, coding for the enzymes involved in the indolocarbazole aglycone biosynthesis, two genes, staG and staN, for the bond formation between the aglycone and deoxysugar, eight genes, staA, staB, staE, staJ, staI, staK, staMA, and staMB, for the deoxysugar biosynthesis and one gene, staR is a transcriptional regulator. Heterologous gene expression of a 38-kb fragment containing a complete set of the biosynthetic genes for staurosporine cloned into pTOYAMAcos confirmed its role in staurosporine biosynthesis. Moreover, the distribution of the gene for chromopyrrolic acid synthase, the key enzyme for the biosynthesis of indolocarbazole aglycone, in actinomycetes was investigated, and rebD homologs were shown to exist only in the strains producing indolocarbazole antibiotics.

  5. The Inhibition and Resistance Mechanisms of Actinonin, Isolated from Marine Streptomyces sp. NHF165, against Vibrio anguillarum

    PubMed Central

    Yang, Na; Sun, Chaomin

    2016-01-01

    Vibrio sp. is the most serious pathogen in marine aquaculture, and the development of anti-Vibrio agents is urgently needed. However, it is extreme lack of high-throughput screening (HTS) model for searching anti-Vibrio compounds. Here, we established a protein-based HTS screening model to identify agents targeting peptide deformylase (PDF) of Vibrio anguillarum. To find potential anti-Vibrio compounds, crude extracts derived from marine actinomycetes were applied for screening with this model. Notably, crude extract of strain Streptomyces sp. NHF165 inhibited dramatically both on V. anguillarum PDF (VaPDF) activity and V. anguillarum cell growth. And actinonin was further identified as the functional component. Anti-VaPDF and anti-V. anguillarum activities of actinonin were dose-dependent, and the IC50 values were 6.94 and 2.85 μM, respectively. To understand the resistance of V. anguillarum against actinonin, spontaneous V. anguillarum mutants with resistance against actinonin were isolated. Surprisingly, for the resistant strains, the region between 774 and 852 base pairs was found to be absent in the gene folD which produces 10-formyl-tetrahydrofolate, a donor of N-formyl to Met-tRNAfmet. When compared to the wild type strain, ΔfolD mutant showed eight times of minimum inhibition concentration on actinonin, however, the folD complementary strain could not grow on the medium supplemented with actinonin, which suggested that folD gene mutation was mainly responsible for the actinonin resistance. To our knowledge, this is the first report showing that marine derived Streptomyces sp. could produce actinonin with anti-VaPDF activity and the resistance against actinonin by V. anguillarum is mediated by mutation in folD gene. PMID:27679625

  6. Cytotoxic Activity of Bioactive Compound 1, 2- Benzene Dicarboxylic Acid, Mono 2- Ethylhexyl Ester Extracted from a Marine Derived Streptomyces sp. VITSJK8.

    PubMed

    Krishnan, Kannabiran; Mani, Abirami; Jasmine, Subashini

    2014-01-01

    Marine Streptomyces are prolific producers of majority of bioactive secondary metabolites which are used in pharmaceutical industry as effective drugs against life threatening diseases. The cytotoxic activity of the pure compound 1, 2- benzene dicarboxylic acid, mono 2- ethylhexyl ester (DMEHE) from marine derived actinomycete Streptomyces sp. VITSJK8 was investigated against mouse embryonic fibroblast (NIH 3T3) and human keratinocyte (HaCaT) normal cell lines, human hepatocellular liver carcinoma (HepG 2) and human breast adenocarcinoma (MCF-7) cell lines by using MTT assay. The compound DMEHE exhibited IC 50 values of 42, 100, 250 and 500 µg/ ml against HepG2, MCF-7, HaCaT and NIH 3T3 cell lines, respectively. The effect of DMEHE on the growth of cancer cell lines was expressed as the % of viability. Cell viability was recorded as 67.7%, 78.14%, 82.23% and 96. 11% in HepG2, MCF-7, HaCaT and NIH 3T3 cells, respectively. The results of the study conclude that the bioactive compound isolated from the potential isolate Streptomyces sp. VITSJK8 exhibited cytotoxic activity against HepG2 and MCF- 7 cancer cell lines and low toxicity against normal HaCaT and NIH 3T3 cell lines.

  7. Cytotoxic Activity of Bioactive Compound 1, 2- Benzene Dicarboxylic Acid, Mono 2- Ethylhexyl Ester Extracted from a Marine Derived Streptomyces sp. VITSJK8

    PubMed Central

    Krishnan, Kannabiran; Mani, Abirami; Jasmine, Subashini

    2014-01-01

    Marine Streptomyces are prolific producers of majority of bioactive secondary metabolites which are used in pharmaceutical industry as effective drugs against life threatening diseases. The cytotoxic activity of the pure compound 1, 2- benzene dicarboxylic acid, mono 2- ethylhexyl ester (DMEHE) from marine derived actinomycete Streptomyces sp. VITSJK8 was investigated against mouse embryonic fibroblast (NIH 3T3) and human keratinocyte (HaCaT) normal cell lines, human hepatocellular liver carcinoma (HepG 2) and human breast adenocarcinoma (MCF-7) cell lines by using MTT assay. The compound DMEHE exhibited IC 50 values of 42, 100, 250 and 500 µg/ ml against HepG2, MCF-7, HaCaT and NIH 3T3 cell lines, respectively. The effect of DMEHE on the growth of cancer cell lines was expressed as the % of viability. Cell viability was recorded as 67.7%, 78.14%, 82.23% and 96. 11% in HepG2, MCF-7, HaCaT and NIH 3T3 cells, respectively. The results of the study conclude that the bioactive compound isolated from the potential isolate Streptomyces sp. VITSJK8 exhibited cytotoxic activity against HepG2 and MCF- 7 cancer cell lines and low toxicity against normal HaCaT and NIH 3T3 cell lines. PMID:25635251

  8. Streptomyces mexicanus sp. nov., a xylanolytic micro-organism isolated from soil.

    PubMed

    Petrosyan, Pavel; García-Varela, Martin; Luz-Madrigal, Agustín; Huitrón, Carlos; Flores, María Elena

    2003-01-01

    The taxonomic position of a thermophilic actinomycete strain isolated from soil was examined using a polyphasic approach. The strain, designated CH-M-1035T, was assigned to the genus Streptomyces on the basis of chemical and morphological criteria. It formed Rectiflexibiles aerial hyphae that carried long chains of rounded, smooth spores. The almost complete nucleotide sequence of the 16S rRNA gene of strain CH-M-1035T was determined and its comparison with the 16S rDNA sequences of previously studied streptomycetes confirmed the assignment of the novel strain to the genus Streptomyces. Strain CH-M-1035T clustered with species belonging to the Streptomyces thermodiastaticus clade in the 1 6S-rDNA-based phylogenetic tree. However, the phenotypic properties of strain CH-M-1035T differed from those of the recognized species within this clade. Therefore, it is proposed that strain CH-M-1035T be classified as a novel species within the genus Streptomyces, as Streptomyces mexicanus (type strain CH-M-1035T =DSM 41796T =BM-B-384T =NRRL B-24196T).

  9. An endophytic Streptomyces sp. strain DHV3-2 from diseased root as a potential biocontrol agent against Verticillium dahliae and growth elicitor in tomato (Solanum lycopersicum).

    PubMed

    Cao, Peng; Liu, Chongxi; Sun, Pengyu; Fu, Xuepeng; Wang, Shaoxian; Wu, Fengzhi; Wang, Xiangjing

    2016-12-01

    Plant endophytes play important roles in biocontrol of plant diseases. Actinomycetes are used for biocontrol of fungal diseases caused by Verticillium dahliae. Many studies have focused on the endophytic actinomycetes isolated from the roots of healthy plants, but few on those from the roots of diseased plants. In the present research, actinomycetes were isolated from the roots of diseased and healthy tomato plants, respectively. The results showed that, in total, 86 endophytic actinomycetes were isolated for screening of their antimicrobial activities, 8 of which showed antagonism to V. dahliae in vitro. Among the 8 antagonistic strains, 5 (out of 36) were from the roots of diseased plants, with inhibition diameter zones ranging from 11.2 to 18.2 mm, whereas 3 (out of 50) were from the roots of healthy plants, with inhibition diameter zones ranging from 11.5 to 15.5 mm. Endophytic strain DHV3-2 was isolated from the root of a diseased plant and demonstrated a potent effect against V. dahliae and other pathogenic fungi by showing the largest inhibition diameter zones among all the eight antagonistic strains. Thus, strain DHV3-2 was chosen to investigate its biological control efficacies in vivo. Further study showed that the disease incidence and disease severity indices of tomato Verticillium wilt decreased significantly (P < 0.05). We also found that the plant shoot fresh weight and height increased greatly (P < 0.05) upon treatment with strain DHV3-2 compared to the plants uninoculated in greenhouse conditions. Root colonization showed that strain DHV3-2 had the higher root-colonizing capacity in the roots of infected plants compared with the roots of healthy plants. This isolate was identified as Streptomyces sp. based on morphological characteristics and 16S rRNA gene analysis. In conclusion, the roots of diseased tomato plants are a potential reservoir of biological control actinomycetes, and Streptomyces sp. strain DHV3-2 is a potential biocontrol

  10. Identification of actinomycete communities in Antarctic soil from Barrientos Island using PCR-denaturing gradient gel electrophoresis.

    PubMed

    Learn-Han, L; Yoke-Kqueen, C; Shiran, M S; Vui-Ling, C M W; Nurul-Syakima, A M; Son, R; Andrade, H M

    2012-02-08

    The diversity of specific bacteria taxa, such as the actinomycetes, has not been reported from the Antarctic island of Barrientos. The diversity of actinomycetes was estimated with two different strategies that use PCR-denaturing gradient gel electrophoresis. First, a PCR was applied, using a group-specific primer that allows selective amplification of actinomycete sequences. Second, a nested-PCR approach was used that allows the estimation of the relative abundance of actinomycetes within the bacterial community. Molecular identification, which was based on 16S rDNA sequence analysis, revealed eight genera of actinomycetes, Actinobacterium, Actinomyces, an uncultured Actinomycete, Streptomyces, Leifsonia, Frankineae, Rhodococcus, and Mycobacterium. The uncultured Actinomyces sp and Rhodococcus sp appear to be the prominent genera of actinomycetes in Barrientos Island soil. PCR-denaturing gradient gel electrophoresis patterns were used to look for correlations between actinomycete abundance and environmental characteristics, such as type of rookery and vegetation. There was a significant positive correlation between type of rookery and abundance of actinomycetes; soil samples collected from active chinstrap penguin rookeries had the highest actinomycete abundance. Vegetation type, such as moss, which could provide a microhabitat for bacteria, did not correlate significantly with actinomycete abundance.

  11. An ammonium sulfate sensitive chitinase from Streptomyces sp. CS501.

    PubMed

    Rahman, Md Arifur; Choi, Yun Hee; Pradeep, G C; Yoo, Jin Cheol

    2014-12-01

    A chitinase from Streptomyces sp. CS501 was isolated from the Korean soil sample, purified by single-step chromatography, and biochemically characterized. The extracellular chitinase (Ch501) was purified to 4.60 fold with yield of 28.74 % using Sepharose Cl-6B column. The molecular mass of Ch501 was approximately 43 kDa as estimated by SDS-PAGE and zymography. The enzyme (Ch501) was found to be stable over a broad pH range (5.0-10.0) and temperature (up to 50 °C), and have an optimum temperature of 60 °C. N-terminal sequence of Ch501 was AAYDDAAAAA. Intriguingly, Ch501 was highly sensitive to ammonium sulfate but it's completely suppressed activity was recovered after desalting out. TLC analysis of Ch501 showed the production of N-acetyl D-glucosamine (GlcNAc) and Diacetylchitobiose (GlcNAc)2, as a principal hydrolyzed product. Ch501 shows antifungal activity against Fusarium solani and Aspergillus brasiliensis, which can be used for the biological control of fungus. As has been simple in purification, stable in a broad range of pH, ability to produce oligosaccharides, and antifungal activity showed that Ch501 has potential applications in industries as for chitooligosaccharides production used as prebiotics and/or for the biological control of plant pathogens in agriculture.

  12. Switching antibiotics production on and off in actinomycetes by an IclR family transcriptional regulator from Streptomyces peucetius ATCC 27952.

    PubMed

    Chaudhary, Amit Kumar; Singh, Bijay; Maharjan, Sushila; Jha, Amit Kumar; Kim, Byung-Gee; Sohng, Jae Kyung

    2014-08-01

    Doxorubicin, produced by Streptomyces peucetius ATCC 27952, is tightly regulated by dnrO, dnrN, and dnrI regulators. Genome mining of S. peucetius revealed the presence of the IclR (doxR) type family of transcription regulator mediating the signal-dependent expression of operons at the nonribosomal peptide synthetase gene cluster. Overexpression of doxR in native strain strongly repressed the drug production. Furthermore, it also had a negative effect on the regulatory system of doxorubicin, wherein the transcript of dnrI was reduced to the maximum level in comparision with the other two. Interestingly, the overexpression of the same gene also had strong inhibitory effects on the production of actinorhodin (blue pigment) and undecylprodigiosin (red pigment) in Streptomyces coelicolor M145, herboxidiene production in Streptomyces chromofuscus ATCC 49982, and spinosyn production in Saccharopolyspora spinosa NRRL 18395, respectively. Moreover, DoxR exhibited pleiotropic effects on the production of blue and red pigments in S. coelicolor when grown in different agar media, wherein the production of blue pigment was inhibited in R2YE medium and the red pigment was inhibited in YEME medium. However, the production of both blue and red pigments from S. coelicolor harboring doxR was halted in ISP2 medium, whereas S. coelicolor produced both pigmented antibiotics in the same plate. These consequences demonstrate that the on and off production of these antibiotics was not due to salt stress or media compositions, but was selectively controlled in actinomycetes.

  13. Screening of mutant strain Streptomyces mediolani sp. AC37 for (-)-8-O-methyltetrangomycin production enhancement.

    PubMed

    Jiménez, Jakeline Trejos; Sturdíková, Maria; Brezová, Vlasta; Svajdlenka, Emil; Novotová, Marta

    2012-12-01

    Streptomyces mediolani sp. AC37 was isolated from the root system of higher plant Taxus baccata and produced metabolite identified as (-)-8-O-methyltetrangomycin according to LC/MS/MS analysis. In our screening program for improvements of bioactive secondary metabolites from plant associate streptomycetes, mutation was used as a tool for the induction of genetic variations for selection of higher (-)-8-O-methyltetrangomycin producers of isolates. S. mediolani sp. AC37 was treated with UV irradiation and chemical mutagenic treatment (N-nitroso-N-methyl-urea). The radical scavenging and antioxidant capacity of (-)-8-O-methyltetrangomycin and extracts isolated from mutants were tested using EPR spin trapping technique and ABTS(·+) assay. Comparison of electron microscopic images of Streptomyces sp. AC37 and mutant strains of Streptomyces sp. AC37 revealed substantial differences in morphology and ultrastructure.

  14. Actinopolyspora algeriensis sp. nov., a novel halophilic actinomycete isolated from a Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2012-09-01

    A halophilic actinomycete strain designated H19(T), was isolated from a Saharan soil in the Bamendil region (Ouargla province, South Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Actinopolyspora, and 16S rRNA gene sequence analysis confirmed that strain H19(T) was a novel species of the genus Actinopolyspora. DNA-DNA hybridization value between strain H19(T) and the nearest Actinopolyspora species, A. halophila, was clearly below the 70 % threshold. The genotypic and phenotypic data showed that the organism represents a novel species of the genus Actinopolyspora for which the name Actinopolyspora algeriensis sp. nov. is proposed, with the type strain H19(T) (= DSM 45476(T) = CCUG 62415(T)).

  15. Nocardia kroppenstedtii sp. nov., an actinomycete isolated from a lung transplant patient with a pulmonary infection.

    PubMed

    Jones, Amanda L; Fisher, Andrew J; Mahida, Rahul; Gould, Kate; Perry, John D; Hannan, Margaret M; Judge, Eoin P; Brown, Ros; Boagey, Kimberley; Goodfellow, Michael

    2014-03-01

    A novel actinomycete, strain N1286(T), isolated from a lung transplant patient with a pulmonary infection, was provisionally assigned to the genus Nocardia. The strain had chemotaxonomic and morphological properties typical of members of the genus Nocardia and formed a distinct phyletic line in the Nocardia 16S rRNA gene tree. Isolate N1286(T) was most closely related to Nocardia farcinica DSM 43665(T) (99.8% gene sequence similarity) but could be distinguished from the latter by the low level of DNA-DNA relatedness. These strains were also distinguishable on the basis of a broad range of phenotypic properties. It is concluded that strain N1286(T) represents a novel species of the genus Nocardia for which the name Nocardia kroppenstedtii sp. nov. is proposed. The type strain is N1286(T) ( = DSM 45810(T) = NCTC 13617(T)).

  16. Genome Sequence of Streptomyces sp. H-KF8, a Marine Actinobacterium Isolated from a Northern Chilean Patagonian Fjord.

    PubMed

    Undabarrena, Agustina; Ugalde, Juan Antonio; Castro-Nallar, Eduardo; Seeger, Michael; Cámara, Beatriz

    2017-02-09

    Streptomyces sp. H-KF8 is a fjord-derived marine actinobacterium capable of producing antimicrobial activity. Streptomyces sp. H-KF8 was isolated from sediments of the Comau fjord, located in the northern Chilean Patagonia. Here, we report the 7.7-Mb genome assembly, which represents the first genome of a Chilean marine actinobacterium.

  17. Genome Sequence of Streptomyces sp. H-KF8, a Marine Actinobacterium Isolated from a Northern Chilean Patagonian Fjord

    PubMed Central

    Undabarrena, Agustina; Ugalde, Juan Antonio; Castro-Nallar, Eduardo; Seeger, Michael

    2017-01-01

    ABSTRACT Streptomyces sp. H-KF8 is a fjord-derived marine actinobacterium capable of producing antimicrobial activity. Streptomyces sp. H-KF8 was isolated from sediments of the Comau fjord, located in the northern Chilean Patagonia. Here, we report the 7.7-Mb genome assembly, which represents the first genome of a Chilean marine actinobacterium. PMID:28183776

  18. Characterization and Optimization of Biosynthesis of Bioactive Secondary Metabolites Produced by Streptomyces sp. 8812.

    PubMed

    Rajnisz, Aleksandra; Guśpiel, Adam; Postek, Magdalena; Ziemska, Joanna; Laskowska, Anna; Rabczenko, Daniel; Solecka, Jolanta

    2016-01-01

    The nutritional requirements and environmental conditions for a submerged culture of Streptomyces sp. 8812 were determined. Batch and fed-batch Streptomyces sp. 8812 fermentations were conducted to obtain high activity of secondary metabolites. In the study several factors were examined for their influence on the biosynthesis of the active metabolites-7-hydroxy-6-oxo-2,3,4,6-tetrahydroisoquinoline-3-carboxy acid (C10H9NO4) and N-acetyl-3,4-dihydroxy-L-phenylalanine (C11H13NO5): changes in medium composition, pH of production medium, various growth phases of seed culture, amino acid supplementation and addition of anion exchange resin to the submerged culture. Biological activities of secondary metabolites were examined with the use of DD-carboxypeptidase 64-575 and horseradish peroxidase. Streptomyces sp. 8812 mycelium was evaluated under fluorescent microscopy and respiratory activity of the strain was analyzed. Moreover, the enzymatic profiles of the strain with the use of Api ZYM test were analyzed and genetic analysis made. Phylogenetic analysis of Streptomyces sp. 8812 revealed that its closest relative is Streptomyces capoamus JCM 4734 (98%), whereas sequence analysis for 16S rRNA gene using NCBI BLAST algorithm showed 100% homology between these two strains. Biosynthetic processes, mycelium growth and enzyme inhibitory activities of these two strains were also compared.

  19. Antibacterial activity of Pseudonocardia sp. JB05, a rare salty soil actinomycete against Staphylococcus aureus.

    PubMed

    Jafari, Nesa; Behroozi, Reza; Farajzadeh, Davoud; Farsi, Mohammad; Akbari-Noghabi, Kambiz

    2014-01-01

    Staphylococcus aureus is a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds against S. aureus and characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated as Pseudonocardia sp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL(-1) against S. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain of Pseudonocardia. The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

  20. Submerged culture screening of two strains of Streptomyces sp. with high keratinolytic activity.

    PubMed

    Garcia-Kirchner, O; Bautista-Ramirez, M E; Segura-Granados, M

    1998-01-01

    Keratinases can be used for the production of potentially important hydrolyzed proteins and chemicals. This study investigated the keratinolytic activity of Streptomyces sp on keratinaceous materials like wool. High levels of proteolytic and keratinolytic activity were obtained after 96 h of culture when two Streptomyces sp strains were grown on basal medium containing mineral salts and 3% (w/v) of defatted wool as a source of energy, carbon, and nitrogen. The cell-free culture filtrates exhibited rapid proteolytic digestion of keratin powder. Currently, the authors are testing whether the enzymatic activity obtained is in fact keratinolytic, and not only an alkaline protease activity.

  1. Streptomyces aridus sp. nov., isolated from a high altitude Atacama Desert soil and emended description of Streptomyces noboritoensis Isono et al. 1957.

    PubMed

    Idris, Hamidah; Labeda, David P; Nouioui, Imen; Castro, Jean Franco; Del Carmen Montero-Calasanz, Maria; Bull, Alan T; Asenjo, Juan A; Goodfellow, Michael

    2017-02-09

    A polyphasic study was undertaken to determine the taxonomic status of a Streptomyces strain which had been isolated from a high altitude Atacama Desert soil and shown to have bioactive properties. The strain, isolate H9(T), was found to have chemotaxonomic, cultural and morphological properties that place it in the genus Streptomyces. 16S rRNA gene sequence analyses showed that the isolate forms a distinct branch at the periphery of a well-delineated subclade in the Streptomyces 16S rRNA gene tree together with the type strains of Streptomyces crystallinus, Streptomyces melanogenes and Streptomyces noboritoensis. Multi-locus sequence analysis (MLSA) based on five house-keeping gene alleles showed that isolate H9(T) is closely related to the latter two type strains and to Streptomyces polyantibioticus NRRL B-24448(T). The isolate was distinguished readily from the type strains of S. melanogenes, S. noboritoensis and S. polyantibioticus using a combination of phenotypic properties. Consequently, the isolate is considered to represent a new species of Streptomyces for which the name Streptomyces aridus sp. nov. is proposed; the type strain is H9(T) (=NCIMB 14965(T)=NRRL B65268(T)). In addition, the MLSA and phenotypic data show that the S. melanogenes and S. noboritoensis type strains belong to a single species, it is proposed that S. melanogenes be recognised as a heterotypic synonym of S. noboritoensis for which an emended description is given.

  2. Antibiotics production by an actinomycete isolated from the termite gut.

    PubMed

    Matsui, Toru; Tanaka, Junichi; Namihira, Tomoyuki; Shinzato, Naoya

    2012-12-01

    As well as the search for new antibiotics, a new resource or strains for the known antibiotics is also important. Microbial symbionts in the gut of termites could be regarded as one of the feasible resource for such purpose. In this study, antibiotic-producing actinomycetes were screened from symbionts of the termite gut. 16SrRNA sequence analysis for the 10 isolates revealed that they belong to actinomycetes such as Streptomyces sp., Kitasatospora sp., and Mycobacterium sp. A culture broth from one of the isolate, namely strain CA1, belonging to the genera Streptomyces exhibited antagonistic activity against actinomycetes (Micrococcus spp.), gram-positive bacteria (Bacillus spp.), and yeast (Candida spp.). The structures of 2 compounds isolated from the culture broth of the strain CA1 were identified as those of actinomycin X2 and its analog, D. This study is the first to report that some symbionts of the termite gut are antibiotic-producing actinomycetes, and suggest that the termite gut is a feasible resource for bioprospecting.

  3. Discovery of C-Glycosylpyranonaphthoquinones in Streptomyces sp. MBT76 by a Combined NMR-Based Metabolomics and Bioinformatics Workflow.

    PubMed

    Wu, Changsheng; Du, Chao; Ichinose, Koji; Choi, Young Hae; van Wezel, Gilles P

    2017-02-24

    Mining of microbial genomes has revealed that actinomycetes harbor far more biosynthetic potential for bioactive natural products than anticipated. Activation of (cryptic) biosynthetic gene clusters and identification of the corresponding metabolites has become a focal point for drug discovery. Here, we applied NMR-based metabolomics combined with bioinformatics to identify novel C-glycosylpyranonaphthoquinones in Streptomyces sp. MBT76 and to elucidate the biosynthetic pathway. Following activation of the cryptic qin gene cluster for a type II polyketide synthase (PKS) by constitutive expression of its pathway-specific activator, bioinformatics coupled to NMR profiling facilitated the chromatographic isolation and structural elucidation of qinimycins A-C (1-3). The intriguing structural features of the qinimycins, including 8-C-glycosylation, 5,14-epoxidation, and 13-hydroxylation, distinguished these molecules from the model pyranonaphthoquinones actinorhodin, medermycin, and granaticin. Another novelty lies in the unusual fusion of a deoxyaminosugar to the pyranonaphthoquinone backbone during biosynthesis of the antibiotics BE-54238 A and B (4, 5). Qinimycins showed weak antimicrobial activity against Gram-positive bacteria. Our work shows the utility of combining bioinformatics, targeted activation of cryptic gene clusters, and NMR-based metabolic profiling as an effective pipeline for the discovery of microbial natural products with distinctive skeletons.

  4. Genome Sequence of the Mycorrhiza Helper Bacterium Streptomyces sp. Strain AcH 505

    PubMed Central

    Feldhahn, L.; Buscot, F.; Wubet, T.

    2015-01-01

    A draft genome sequence of Streptomyces sp. strain AcH 505 is presented here. The genome encodes 22 secondary metabolite gene clusters and a large arsenal of secreted proteins, and their comparative and functional analyses will help to advance our knowledge of symbiotic interactions and fungal and plant biomass degradation. PMID:25838498

  5. Draft Genome Sequence of an Anicemycin Producer, Streptomyces sp. TP-A0648

    PubMed Central

    Hosoyama, Akira; Kimura, Akane; Ichikawa, Natsuko; Igarashi, Yasuhiro

    2017-01-01

    ABSTRACT We report the draft genome sequence of Streptomyces sp. TP-A0648 isolated from a leaf of Aucuba japonica. This strain produces a new tumor cell growth inhibitor designated anicemycin. The genome harbors at least 12 biosynthetic gene clusters for polyketides and nonribosomal peptides, suggesting the potential to produce diverse secondary metabolites. PMID:28082502

  6. Production of induced secondary metabolites by a co-culture of sponge-associated actinomycetes, Actinokineospora sp. EG49 and Nocardiopsis sp. RV163.

    PubMed

    Dashti, Yousef; Grkovic, Tanja; Abdelmohsen, Usama Ramadan; Hentschel, Ute; Quinn, Ronald J

    2014-05-22

    Two sponge-derived actinomycetes, Actinokineospora sp. EG49 and Nocardiopsis sp. RV163, were grown in co-culture and the presence of induced metabolites monitored by ¹H NMR. Ten known compounds, including angucycline, diketopiperazine and β-carboline derivatives 1-10, were isolated from the EtOAc extracts of Actinokineospora sp. EG49 and Nocardiopsis sp. RV163. Co-cultivation of Actinokineospora sp. EG49 and Nocardiopsis sp. RV163 induced the biosynthesis of three natural products that were not detected in the single culture of either microorganism, namely N-(2-hydroxyphenyl)-acetamide (11), 1,6-dihydroxyphenazine (12) and 5a,6,11a,12-tetrahydro-5a,11a-dimethyl[1,4]benzoxazino[3,2-b][1,4]benzoxazine (13a). When tested for biological activity against a range of bacteria and parasites, only the phenazine 12 was active against Bacillus sp. P25, Trypanosoma brucei and interestingly, against Actinokineospora sp. EG49. These findings highlight the co-cultivation approach as an effective strategy to access the bioactive secondary metabolites hidden in the genomes of marine actinomycetes.

  7. Enhanced polyaromatic hydrocarbon degradation by adapted cultures of actinomycete strains.

    PubMed

    Bourguignon, Natalia; Isaac, Paula; Alvarez, Héctor; Amoroso, María J; Ferrero, Marcela A

    2014-12-01

    Fifteen actinomycete strains were evaluated for their potential use in removal of polycyclic aromatic hydrocarbons (PAH). Their capability to degrade of naphthalene, phenanthrene, and pyrene was tested in minimal medium (MM) and MM with glucose as another substrate. Degradation of naphthalene in MM was observed in all isolates at different rates, reaching maximum values near to 76% in some strains of Streptomyces, Rhodococcus sp. 016 and Amycolatopsis tucumanensis DSM 45259. Maximum values of degradation of phenanthrene in MM occurred in cultures of A. tucumanensis DSM 45259 (36.2%) and Streptomyces sp. A12 (20%), while the degradation of pyrene in MM was poor and only significant with Streptomyces sp. A12 (4.3%). Because of the poor performance when growing on phenanthrene and pyrene alone, Rhodococcus sp. 20, Rhodococcus sp. 016, A. tucumanensis DSM 45259, Streptomyces sp. A2, and Streptomyces sp. A12 were challenged to an adaptation schedule of successive cultures on a fresh solid medium supplemented with PAHs, decreasing concentration of glucose in each step. As a result, an enhanced degradation of PAHs by adapted strains was observed in the presence of glucose as co-substrate, without degradation of phenanthrene and pyrene in MM while an increase to up to 50% of degradation was seen with these strains in glucose amended media. An internal fragment of the catA gene, which codes for catechol 1,2-dioxygenase, was amplified from both Rhodococcus strains, showing the potential for degradation of aromatic compounds via salycilate. These results allow us to propose the usefulness of these actinomycete strains for PAH bioremediation in the environment.

  8. Streptomyces lacrimifluminis sp. nov., a novel actinobacterium that produces antibacterial compounds, isolated from soil.

    PubMed

    Zhang, Binglin; Tang, Shukun; Chen, Ximing; Zhang, Ling; Zhang, Gaoseng; Zhang, Wei; Liu, Guangxiu; Chen, Tuo; Li, Shiweng; Dyson, Paul

    2016-12-01

    A novel actinobacterial strain, designated Z1027T, was isolated from a soil sample collected near the Tuotuo River, Qinghai-Tibet Plateau (China). The strain exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. The taxonomic position of strain Z1027T was determined using a polyphasic approach. The organism had chemotaxonomic and morphological properties consistent with its classification in the genus Streptomyces and formed a distinct phyletic line in the 16S rRNA gene tree, together with Streptomyces turgidiscabies ATCC 700248T (99.19 % similarity), Streptomyces graminilatus JL-6T (98.84 %) and Streptomyces reticuliscabiei CFBP 4531T (98.36 %). The genomic DNA G+C content of strain Z1027T was 74±1 mol%. The DNA-DNA relatedness values between strain Z1027T and Streptomyces turgidiscabies ATCC 700248T and Streptomyces reticuliscabiei CFBP 4531T were 38.5±0.4 and 26.2±1.2 %, respectively, both of them significantly lower than 70 %. Chemotaxonomic data revealed that strain Z1027T possessed MK-9(H6) and MK-9(H8) as the major menaquinones, ll-diaminopimelic acid as the diagnostic diamino acid and galactose as a whole-cell sugar. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatydilinositol and seven other unknown polar lipids were detected; iso-C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0 were the major fatty acids. On the basis of these genotypic and phenotypic data, it is proposed that isolate Z1027T (=CGMCC 4.7272T=JCM 31054T) should be classified as the type strain of a novel species of the genus Streptomyces,Streptomyces lacrimifluminis sp. nov.

  9. Actinopolyspora mzabensis sp. nov., a halophilic actinomycete isolated from an Algerian Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-10-01

    A halophilic actinomycete strain, designated H55(T), was isolated from Saharan soil sampled in the Mzab region (Ghardaïa, southern Algeria) and was characterized in a taxonomic study using a polyphasic approach. The cell wall was determined to contain meso-diaminopimelic acid and the characteristic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-10(H4) and MK-9(H4). The predominant cellular fatty acids were determined to be anteiso-C17 : 0, iso-C16 : 0 and iso-C15 : 0. The diagnostic phospholipid detected was phosphatidylcholine. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Actinopolyspora, and 16S rRNA gene sequence analysis confirmed that strain H55(T) was a member of this genus. DNA-DNA hybridization values between strain H55(T) and the type strains of the nearest species of the genus Actinopolyspora, Actinopolyspora erythraea and A. alba, were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the organism represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora mzabensis sp. nov. is proposed, with the type strain H55(T) ( = DSM 45460(T) = CCUG 62965(T)).

  10. Saccharothrix saharensis sp. nov., an actinomycete isolated from Algerian Saharan soil.

    PubMed

    Boubetra, Dalila; Zitouni, Abdelghani; Bouras, Noureddine; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-10-01

    The taxonomic position of a novel actinomycete, strain SA152(T), isolated from a sample of Algerian Saharan soil, was determined using a polyphasic taxonomic approach. The strain produced abundant aerial mycelium and fragmented substrate mycelium on most media tested. Chemotaxonomically and phylogenetically, the strain was related to the members of the genus Saccharothrix. Results of 16S rRNA gene sequence comparison revealed that strain SA152(T) shared the highest degree of 16S rRNA gene sequence similarity with Saccharothrix xinjiangensis NBRC 101911(T) (99.3 %) and Saccharothrix texasensis NRRL B-16134(T) (98.9 %). However, DNA-DNA hybridization studies showed only 16.2 % relatedness with S. xinjiangensis DSM 44896(T) and 33.9 % relatedness with S. texasensis DSM 44231(T). Based upon genotypic and phenotypic differences from other members of the genus, a novel species, Saccharothrix saharensis sp. nov., is proposed, with SA152(T) ( = DSM 45456(T) = CCUG 60213(T)) as the type strain.

  11. Prauserella isguenensis sp. nov., a halophilic actinomycete isolated from desert soil.

    PubMed

    Saker, Rafika; Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-05-01

    Two actinomycete strains, designated H225(T) and H137, were isolated from two soil samples collected from the arid region of Ahbas at Béni-Isguen (Mzab), located in the Algerian Sahara. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the novel strains should be assigned to the genus Prauserella of the family Pseudonocardiaceae , and they were therefore subjected to a polyphasic taxonomic study. These two strains contained meso-diaminopimelic acid as the diagnostic diamino acid and arabinose and galactose as major whole-cell sugars. The diagnostic phospholipid was phosphatidylethanolamine. The predominant menaquinone was MK-9(H4), and the major fatty acid was iso-C16 : 0. DNA-DNA hybridization values between strain H225(T) and its closest phylogenetic neighbours, namely Prauserella flava DSM 45265(T), Prauserella alba DSM 44590(T), Prauserella aidingensis DSM 45266(T), Prauserella salsuginis DSM 45264(T) and Prauserella sediminis DSM 45267(T), were clearly below the 70% threshold used for species delineation. The genomic DNA G+C content of strains H225(T) and H137 was 70.4 mol%. On the basis of phenotypic and genotypic data, strains H225(T) and H137(T) are considered to represent a novel species of the genus Prauserella , for which the name Prauserella isguenensis sp. nov. is proposed. The type strain is H225(T) ( =DSM 46664(T) = CECT 8577(T)).

  12. Pseudonocardia antimicrobica sp. nov., a novel endophytic actinomycete associated with Artemisia annua L. (sweet wormwood).

    PubMed

    Zhao, Guo-Zhen; Li, Jie; Qin, Yu-Li; Miao, Cui-Ping; Wei, Da-Qiao; Zhang, Si; Xu, Li-Hua; Li, Wen-Jun

    2012-09-01

    A Gram-reaction-positive, non-motile, endophytic actinomycete, designated strain YIM 63235(T), was isolated from the surface-sterilized stems of Artemisia annua L., and characterized to determine its taxonomic position. The strain YIM 63235(T) formed well-differentiated aerial and substrate mycelia on media tested. The phylogenetic tree based on 16S rRNA gene sequences showed that the new isolate formed a distinct lineage within the genus Pseudonocardia, and the strain YIM 63235(T) was closely related to Pseudonocardia parietis 04-St-002(T) (99.1%). However, DNA-DNA relatedness demonstrated that strain YIM 63235(T) was distinct from the closest phylogenetic neighbor. The chemotaxonomic properties of strain YIM 63235(T) were consistent with those of the genus Pseudonocardia: the diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid and MK-8(H(4)) was the predominant menaquinone. The major fatty acids were iso-C(16:0) and iso-C(16:1) H. The DNA G+C content of strain YIM 63235(T) was 71.0 mol%. On the basis of the phenotypic and phylogenetic distinctiveness, the novel isolate was identified as representing a novel species of the genus Pseudonocardia, for which the name Pseudonocardia antimicrobica sp. nov. (type strain YIM 63235(T) =CCTCC AA 208080(T)=DSM 45303(T)) is proposed.

  13. Actinorugispora endophytica gen. nov., sp. nov., an actinomycete isolated from Daucus carota.

    PubMed

    Liu, Min-Jiao; Zhu, Wen-Yong; Li, Jie; Zhao, Guo-Zhen; Xiong, Zhi; Park, Dong-Jin; Hozzein, Wael N; Kim, Chang-Jin; Li, Wen-Jun

    2015-08-01

    An actinomycete strain, designated YIM 690008T, was isolated from Daucus carota collected from South Korea and its taxonomic position was investigated by using a polyphasic approach. The strain grew well on most media tested and no diffusible pigment was produced. The aerial mycelium formed wrinkled single spores and short spore chains, some of which were branched. The whole-cell hydrolysates contained meso-diaminopimelic acid, glucose, mannose, ribose, galactose and rhamnose. The predominant menaquinones were MK-10(H4), MK-10(H6), MK-10(H8) and MK-10(H2). The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides, some unknown phospholipids, glycolipids and polar lipids. The major fatty acids were i-C16 : 0, ai-C17 : 0 and C18 : 1ω9c. The DNA G+C content of the genomic DNA was 63.1 mol%. Phylogenetic analysis indicated that the isolate belongs to the family Nocardiopsaceae. However, based on phenotypic, chemotaxonomic and genotypic data, it was concluded that strain YIM 690008T represents a novel genus and novel species of the family Nocardiopsaceae, for which the name Actinorugispora endophytica gen. nov., sp. nov. (type strain YIM 690008T = DSM 46770T = JCM 30099T = KCTC 29480T) is proposed.

  14. Pseudonocardia hispaniensis sp. nov., a novel actinomycete isolated from industrial wastewater activated sludge.

    PubMed

    Cuesta, G; Soler, A; Alonso, J L; Ruvira, M A; Lucena, T; Arahal, D R; Goodfellow, M

    2013-01-01

    A novel actinomycete, designated PA3(T), was isolated from an oil refinery wastewater treatment plant, located in Palos de la Frontera, Huelva, Spain, and characterized taxonomically by using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate formed a distinct subclade in the Pseudonocardia tree together with Pseudonocardia asaccharolytica DSM 44247(T). The chemotaxonomic properties of the isolate, for example, the presence of MK-8 (H(4)) as the predominant menaquinone and iso-C(16:0) as the major fatty acid, are consistent with its classification in the genus Pseudonocardia. DNA:DNA pairing experiments between the isolate and the type strain of P. asaccharolytica DSM 44247(T) showed that they belonged to separate genomic species. The two strains were readily distinguished using a combination of phenotypic properties. Consequently, it is proposed that isolate PA3(T) represents a novel species for which the name Pseudonocardia hispaniensis sp. nov. is proposed. The type strain is PA3(T) (= CCM 8391(T) = CECT 8030(T)).

  15. Glycomyces tarimensis sp. nov., an actinomycete isolated from a saline-alkali habitat.

    PubMed

    Lv, Ling-Ling; Zhang, Yue-Feng; Zhang, Li-Li

    2015-05-01

    A novel actinomycete strain, designated TRM 45387(T), was isolated from a saline-alkali soil in Xinjiang Province (40° 22' N 79° 08' E), north-west China. The isolate was characterized using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain TRM 45387(T) belonged to the genus Glycomyces and was closely related to Glycomyces arizonensis DSM 44726(T) (96.59% 16S rRNA gene sequence similarity). The G+C content of the DNA was 71.26 mol%. The isolate contained meso-diaminopimelic acid as the diagnostic diamino acid, and xylose, glucose, galactose, arabinose and ribose as the major whole-cell sugars. The diagnostic phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides. The predominant menaquinone was MK-10(H6). The major fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. On the basis of the evidence from this polyphasic study, a novel species, Glycomyces tarimensis sp. nov., is proposed. The type strain of Glycomyces tarimensis is TRM 45387(T) ( =CCTCC AA 2014007(T) =JCM 30184(T)).

  16. Amycolatopsis thailandensis sp. nov., a poly(L-lactic acid)-degrading actinomycete, isolated from soil.

    PubMed

    Chomchoei, Atchareeya; Pathom-Aree, Wasu; Yokota, Akira; Kanongnuch, Chartchai; Lumyong, Saisamorn

    2011-04-01

    A novel actinomycete that was capable of degrading poly(l-lactic acid), strain CMU-PLA07(T), was isolated from soil in northern Thailand. Strain CMU-PLA07(T) had biochemical, chemotaxonomic, morphological and physiological properties that were consistent with its classification in the genus Amycolatopsis. 16S rRNA gene sequence analysis showed that the isolate formed a phyletic line within the genus Amycolatopsis. Strain CMU-PLA07(T) was most similar to Amycolatopsis coloradensis IMSNU 22096(T) (99.5 % 16S rRNA gene sequence similarity) and Amycolatopsis alba DSM 44262(T) (99.4 %). However, strain CMU-PLA07(T) was distinguishable from the type strains of species of the genus Amycolatopsis on the basis of DNA-DNA relatedness and phenotypic data. Therefore, strain CMU-PLA07(T) is considered to represent a novel species of the genus Amycolatopsis, for which the name Amycolatopsis thailandensis sp. nov. is proposed. The type strain is CMU-PLA07(T) ( = JCM 16380(T) = BCC 38279(T)).

  17. Characterization of dusts collected from swine confinement buildings. [Verticillium sp. ; Actinomycetes

    SciTech Connect

    Donham, K.J.; Scallon, L.J.; Popendorf, W.; Treauhaft, M.W.; Roberts, R.D.

    1986-07-01

    The air in 21 different swine confinement buildings was sampled with 37 mm cassette filters with and without cyclone preselectors and with cascade impactors. Filter results yielded a mean total aerosol of 6.3 mg/m/sup 3/, a mean respirable aerosol of 0.5 mg/m/sup 3/; the geometric mean diameter was 2.9 microns. Cascade impactor measurements revealed a mean total aerosol of 7.6 mg/m/sup 3/, a respirable aerosol of 2.5 mg/m/sup 3/ and a mass median diameter of 9.6 microns. The two major constituents in these aerosols were grain particles and dried fecal matter. The grain particles were larger than fecal particles and proportionately more abundant in finishing buildings where 50 kg-100 kg animals are housed. Therefore the respirable fraction was less in finishing buildings than in farrowing and nursery buildings. Culturing of settled dusts yielded six different mold species, with the highest counts for Verticillium sp. (5 x 10/sup 2/ cfu/mg dry dust) grown at 37/sup 0/C. Thermophilic Actinomycetes and both gram negative and gram positive bacteria were isolated.

  18. Biogenic gold nanotriangles from Saccharomonospora sp., an endophytic actinomycetes of Azadirachta indica A. Juss.

    NASA Astrophysics Data System (ADS)

    Verma, Vijay C.; Anand, Swechha; Ulrichs, Christian; Singh, Santosh K.

    2013-04-01

    Microbial biofabrication is emerging as eco-friendly, simpler, and reproducible alternative to chemical synthesis of metals and semiconductor nanoparticles, allowing generation of rare geometrical forms such as nanotriangles and nanoprisms. Highly confined nanostructures like triangles/prisms are interesting class of nanoparticles due to their unique optical properties exploitable in biomedical diagnostics and biosensors. Here, we report for the first time a single-step biological protocol for the synthesis of gold nanotriangles using extract of endophytic actinomycetes Saccharomonospora sp., isolated from surface sterilized root tissues of Azadirachta indica A. Juss., when incubated with an aqueous solution of chloroaurate ions (AuCl- 4/1 mM). Thin, flat occasionally prismatic gold nanotriangles were produced when aqueous chloroaurate ions reacted with the cell-free extract as well as with the biomass of endophytic Saccharomonospora. It was evidenced from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis that proteins of 42 and 50 kD were involved in biosynthesis as well as in stabilization of the nanoparticles. The particle growth process was monitored by UV-vis spectroscopy, and the morphological characterization was carried out by transmission electron microscopy and atomic force microscopy together with X-ray powder diffractions. Although the exact mechanism for this shape-oriented synthesis is not clear so far, the possibility of achieving nanoparticle shape control in a microbial system is exciting.

  19. Glycomyces endophyticus sp. nov., an endophytic actinomycete isolated from the root of Carex baccans Nees.

    PubMed

    Qin, Sheng; Wang, Hai-Bin; Chen, Hua-Hong; Zhang, Yu-Qin; Jiang, Cheng-Lin; Xu, Li-Hua; Li, Wen-Jun

    2008-11-01

    An actinomycete, designated strain YIM 56134(T), was isolated from the root of a Chinese medicinal plant, Carex baccans Nees, collected from Yunnan, south-west China, and subjected to a polyphasic taxonomic study. An analysis of 16S rRNA gene sequence similarities showed that strain YIM 56134(T) was a member of the genus Glycomyces, being most closely related to Glycomyces algeriensis NRRL B-16327(T) (99.0 % similarity), Glycomyces lechevalierae DSM 44724(T) (99.0 %), Glycomyces rutgersensis IFO 14488(T) (98.9 %) and Glycomyces harbinensis IFO 14487(T) (98.7 %). Strain YIM 56134(T) could be distinguished from other established Glycomyces species on the basis of relatively low sequence similarity (<97 %). Phenotypic and chemotaxonomic data supported the affiliation of this strain to the genus Glycomyces. The results of DNA-DNA hybridization and some physiological and biochemical tests allowed differentiation of the strain from related Glycomyces species. Therefore, strain YIM 56134(T) represents a novel species of the genus Glycomyces, for which the name Glycomyces endophyticus sp. nov. is proposed. The type strain is YIM 56134(T) (=KCTC 19152(T) =DSM 45002(T)).

  20. Actinomadura gamaensis sp. nov., a novel actinomycete isolated from soil in Gama, Chad.

    PubMed

    Abagana, Adam Yacoub; Sun, Pengyu; Liu, Chongxi; Cao, Tingting; Zheng, Weiwei; Zhao, Shanshan; Xiang, Wensheng; Wang, Xiangjing

    2016-06-01

    A novel single spore-producing actinomycete, designated strain NEAU-Gz5(T), was isolated from a soil sample from Gama, Chad. A polyphasic taxonomic study was carried out to establish the status of this strain. The diamino acid present in the cell wall is meso-diaminopimelic acid. Glucose, mannose and madurose occur in whole cell hydrolysates. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside and an unidentified glycolipid. The predominant menaquinones were identified as MK-9(H8) and MK-9(H6). The predominant cellular fatty acids were found to be C16:0, iso-C15:0, iso-C16:0 and C18:0 10-methyl. Phylogenetic analysis based on the 16S rRNA gene showed that strain NEAU-Gz5(T) belongs to the genus Actinomadura and is closely related to Actinomadura oligospora JCM 10648(T) (ATCC 43269(T); 98.3 % similarity). However, the low level of DNA-DNA relatedness and some different phenotypic characteristics allowed the strain to be distinguished from its close relatives. Therefore, it is concluded that strain NEAU-Gz5(T) represents a novel species of the genus of Actinomadura, for which the name Actinomadura gamaensis sp. nov. is proposed. The type strain is NEAU-Gz5(T) (= CGMCC 4.7301(T) = DSM 100815(T)).

  1. Nonomuraea solani sp. nov., an actinomycete isolated from eggplant root (Solanum melongena L.).

    PubMed

    Wang, Xiangjing; Zhao, Junwei; Liu, Chongxi; Wang, Jidong; Shen, Yue; Jia, Feiyu; Wang, Liang; Zhang, Ji; Yu, Chao; Xiang, Wensheng

    2013-07-01

    A novel actinomycete, designated strain NEAU-Z6(T), was isolated from eggplant (Solanum melongena L.) root. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain NEAU-Z6(T) belonged to the genus Nonomuraea, with highest sequence similarity to Nonomuraea monospora PT 708(T) (98.83 %), Nonomuraea rosea GW 12687(T) (98.55 %) and Nonomuraea rhizophila YIM 67092(T) (98.02 %). Sequence similarities between strain NEAU-Z6(T) and other species of the genus Nonomuraea ranged from 97.94 % (Nonomuraea candida HMC10(T)) to 96.30 % (Nonomuraea wenchangensis 210417(T)). Key morphological, physiological and chemotaxonomic characteristics of strain NEAU-Z6(T) were congruent with the description of the genus Nonomuraea. The G+C content of the genomic DNA was 64.51 mol%. DNA-DNA relatedness and comparative analysis of physiological, biochemical and chemotaxonomic data allowed genotypic and phenotypic differentiation of strain NEAU-Z6(T) from closely related species. Thus, strain NEAU-Z6(T) represents a novel species of the genus Nonomuraea, for which the name Nonomuraea solani sp. nov. is proposed. The type strain is NEAU-Z6(T) ( = CGMCC 4.7037(T) = DSM 45729(T)).

  2. Metabolomic Profiling and Genomic Study of a Marine Sponge-Associated Streptomyces sp

    PubMed Central

    Viegelmann, Christina; Margassery, Lekha Menon; Kennedy, Jonathan; Zhang, Tong; O’Brien, Ciarán; O’Gara, Fergal; Morrissey, John P.; Dobson, Alan D. W.; Edrada-Ebel, RuAngelie

    2014-01-01

    Metabolomics and genomics are two complementary platforms for analyzing an organism as they provide information on the phenotype and genotype, respectively. These two techniques were applied in the dereplication and identification of bioactive compounds from a Streptomyces sp. (SM8) isolated from the sponge Haliclona simulans from Irish waters. Streptomyces strain SM8 extracts showed antibacterial and antifungal activity. NMR analysis of the active fractions proved that hydroxylated saturated fatty acids were the major components present in the antibacterial fractions. Antimycin compounds were initially putatively identified in the antifungal fractions using LC-Orbitrap. Their presence was later confirmed by comparison to a standard. Genomic analysis of Streptomyces sp. SM8 revealed the presence of multiple secondary metabolism gene clusters, including a gene cluster for the biosynthesis of the antifungal antimycin family of compounds. The antimycin gene cluster of Streptomyces sp. SM8 was inactivated by disruption of the antimycin biosynthesis gene antC. Extracts from this mutant strain showed loss of antimycin production and significantly less antifungal activity than the wild-type strain. Three butenolides, 4,10-dihydroxy-10-methyl-dodec-2-en-1,4-olide (1), 4,11-dihydroxy-10-methyl-dodec-2-en-1,4-olide (2), and 4-hydroxy-10-methyl-11-oxo-dodec-2-en-1,4-olide (3) that had previously been reported from marine Streptomyces species were also isolated from SM8. Comparison of the extracts of Streptomyces strain SM8 and its host sponge, H. simulans, using LC-Orbitrap revealed the presence of metabolites common to both extracts, providing direct evidence linking sponge metabolites to a specific microbial symbiont. PMID:24893324

  3. Draft Genome Sequence of Streptomyces sp. Strain PTY087I2, Isolated from Styela canopus, a Panamanian Tunicate

    PubMed Central

    Gromek, Samantha M.; Sung, Anne A.

    2016-01-01

    Streptomyces sp. PTY087I2 is a marine bacterium isolated from Styela canopus, a tunicate collected in Bocas del Toro, Panama. Here, we report a draft genome sequence for this bacterium, found to have 94.7% average nucleotide identity (ANI) with Streptomyces roseosporus NRRL 11379, and containing a diverse suite of secondary metabolite gene clusters. PMID:27634989

  4. In vitro α-glucosidase inhibition and antioxidative potential of an endophyte species (Streptomyces sp. loyola UGC) isolated from Datura stramonium L.

    PubMed

    Nimal Christhudas, I V S; Praveen Kumar, P; Agastian, P

    2013-07-01

    Endophytic actinomycetes isolated from Datura stramonium L. was evaluated for its effects against in vitro α-glucosidase inhibition, antioxidant, and free radical scavenging activities. Based on microbial cultural characteristic and 16S rRNA sequencing, it was identified as Streptomyces sp. loyola UGC. The methanolic extract of endophytic actinomycetes (MeEA) shows remarkable inhibition of α-glucosidase (IC50 730.21 ± 1.33 μg/ml), scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) (IC50 435.31 ± 1.79 μg/ml), hydroxyl radical (IC50 350.21 ± 1.02 μg/ml), nitric oxide scavenging (IC50 800.12 ± 1.05 μg/ml), superoxide anion radical (IC50 220.31 ± 1.47 μg/ml), as well as a high and dose-dependent reducing power. The MeEA also showed a strong suppressive effect on rat liver lipid peroxidation. Antioxidants of β-carotene linoleate model system revels significantly lower than BHA. The total phenolic content of the extract was 176 mg of catechol equivalents/gram extract. Perusal of this study indicates MeEA can be used as natural resource of α-glucosidase inhibitor and antioxidants.

  5. Characterization of the iron-regulated desA promoter of Streptomyces pilosus as a system for controlled gene expression in actinomycetes.

    PubMed

    Flores, Francisco J; Rincón, Javier; Martín, Juan F

    2003-05-19

    BACKGROUND: The bioavailability of iron is quite low since it is usually present as insoluble complexes. To solve the bioavailability problem microorganisms have developed highly efficient iron-scavenging systems based on the synthesis of siderophores that have high iron affinity. The systems of iron assimilation in microorganisms are strictly regulated to control the intracellular iron levels since at high concentrations iron is toxic for cells. Streptomyces pilosus synthesizes the siderofore desferrioxamine B. The first step in desferrioxamine biosynthesis is decarboxylation of L-lysine to form cadaverine, a desferrioxamine B precursor. This reaction is catalyzed by the lysine decarboxylase, an enzyme encoded by the desA gene that is repressed by iron. RESULTS: The binding of the DmdR (acronym for divalent metal dependent repressor) to the desA promoter in presence of Fe2+ or other divalent ions has been characterized. A 51 bp DNA fragment of the desA promoter containing the 9 bp inverted repeat was sufficient for binding of the DmdR repressor, as observed by the electrophoretic mobility shift assay. The desA mobility shift was prevented by neutralizing DmdR with anti-DmdR antibodies or by chelating the divalent metal in the binding reaction with 2,2'-dipyridyl. Binding to the desA promoter was observed with purified DmdR repressors of Streptomyces coelicolor or Rhodococcus fascians suggesting that there is a common mechanism of iron-regulation in actinomycetes. The complete desA promoter region was coupled using transcriptional fusions to the amy reporter gene (encoding alpha-amylase) in low copy or multicopy Streptomyces vectors. The iron-regulated desA promoter was induced by addition of the iron chelating agent 2,2'-dipyridyl resulting in a strong expression of the reporter gene. CONCLUSIONS: The iron-regulated desA promoter can be used for inducible expression of genes in Streptomyces species, as shown by de-repression of the promoter when coupled to a

  6. Durhamycin, a Pentaene Antifungal Antibiotic from Streptomyces durhamensis sp. n

    PubMed Central

    Gordon, M. A.; Lapa, E. W.

    1966-01-01

    Durhamycin, derived from a previously undescribed soil isolate, is an apparently new antibiotic active against many of the fungi pathogenic for man and animals. Cultural and morphological characteristics and biochemical reactions of Streptomyces durhamensis are detailed. Methods of extraction of the antibiotic, physicochemical properties, antimicrobial spectrum, and results of mouse toxicity and protection tests are given. Images Fig. 1 Fig. 2 Fig. 3 PMID:16349666

  7. Streptomyces yunnanensis sp. nov., a mesophile from soils in Yunnan, China.

    PubMed

    Zhang, Qi; Li, Wen-Jun; Cui, Xiao-Long; Li, Ming-Gang; Xu, Li-Hua; Jiang, Cheng-Lin

    2003-01-01

    A strain was isolated from red soil from the suburb of Kunming in Yunnan, China, during the screening of agricultural antibiotics which prevented and cured wheat-stem rust. This isolate, designated YIM 41004T (= CGMCC 4.1004T = DSM 41793), was identified by a polyphasic approach. The test results suggested that this strain was clearly assigned to the genus Streptomyces and found to be marginally close to Williams cluster 32 based on the morphological and physiological data. The almost-complete 16S rRNA gene sequence of the strain was determined and compared with those of representative streptomycetes. The phylogenetic tree confirmed its membership in the genus Streptomyces and demonstrated that this strain represented a separate phyletic line in a clade encompassed by streptomycetes within cluster 32. Based on the polyphasic evidence, it is therefore proposed that strain YIM 41004T should be classified as Streptomyces yunnanensis sp. nov.

  8. 40 CFR 180.1120 - Streptomyces sp. strain K61; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... FOOD Exemptions From Tolerances § 180.1120 Streptomyces sp. strain K61; exemption from the requirement... of a tolerance in or on all raw agricultural commodities when used as a fungicide for the...

  9. 40 CFR 180.1120 - Streptomyces sp. strain K61; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... FOOD Exemptions From Tolerances § 180.1120 Streptomyces sp. strain K61; exemption from the requirement... of a tolerance in or on all raw agricultural commodities when used as a fungicide for the...

  10. Nocardia halotolerans sp. nov., a halotolerant actinomycete isolated from saline soil.

    PubMed

    Moshtaghi Nikou, Mahdi; Ramezani, Mohaddaseh; Ali Amoozegar, Mohammad; Rasooli, Mehrnoosh; Harirchi, Sharareh; Shahzadeh Fazeli, Seyed Abolhasan; Schumann, Peter; Spröer, Cathrin; Ventosa, Antonio

    2015-09-01

    A novel halotolerant actinomycete, strain Chem15(T), was isolated from soil around Inche-Broun hypersaline wetland; its taxonomic position was determined based on a polyphasic approach. Strain Chem15(T) was strictly aerobic and tolerated NaCl up to 12.5%. The optimum temperature and pH for growth were 28-30 °C and pH 7.0-7.5, respectively. The cell wall of strain Chem15(T) contained meso-diaminopimelic acid as diamino acid and galactose, arabinose and ribose as whole-cell sugars. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The cellular fatty acids profile consisted of C16 : 0, iso-C18 : 0, C18 : 0 10-methyl and C18 : 1ω9c, and the major respiratory quinone was MK-8(H4cycl). The G+C content of the genomic DNA was 68.0 mol%. The novel strain constituted a distinct phyletic line within the genus Nocardia, based on 16S rRNA gene sequence analysis, and was closely associated with Nocardia sungurluensis DSM 45714(T) and Nocardia alba DSM 44684(T) (98.2 and 98.1% 16S rRNA gene sequence similarity, respectively). However DNA-DNA relatedness and phenotypic data demonstrated that strain Chem15(T) was clearly different from closely related species of the genus Nocardia. It is concluded that the organism should be classified as a representative of a novel species of the genus Nocardia, for which the name Nocardia halotolerans sp. nov. is proposed. The type strain is Chem15(T) ( = IBRC-M 10490(T) = LMG 28544(T)).

  11. Micromonospora zeae sp. nov., a novel endophytic actinomycete isolated from corn root (Zea mays L.).

    PubMed

    Shen, Yue; Zhang, Yuejing; Liu, Chongxi; Wang, Xiangjing; Zhao, Junwei; Jia, Feiyu; Yang, Lingyu; Yang, Deguang; Xiang, Wensheng

    2014-11-01

    A novel actinomycete, designated strain NEAU-gq9(T), was isolated from corn root (Zea mays L.) and characterized using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of the genus Micromonospora. On the basis of 16S rRNA gene sequence similarity studies, strain NEAU-gq9(T) was most closely related to Micromonospora zamorensis CR38(T) (99.3%), Micromonospora jinlongensis NEAU-GRX11(T) (99.2%), Micromonospora saelicesensis Lupac 09(T) (99.2%), Micromonospora chokoriensis 2-19(6)(T) (98.9%), Micromonospora coxensis 2-30-b(28)(T) (98.6%) and Micromonospora lupini Lupac 14N(T) (98.5%). Phylogenetic analysis based on the 16S rRNA gene and gyrB gene demonstrated that strain NEAU-gq9(T) is a member of the genus Micromonospora and supported the closest phylogenetic relationship to M. zamorensis CR38(T), M. jinlongensis NEAU-GRX11(T), M. saelicesensis Lupac 09(T), M. chokoriensis 2-19(6)(T) and M. lupini Lupac 14N(T). A combination of DNA-DNA hybridization, morphological and physiological characteristics indicated that the novel strain could be readily distinguished from the closest phylogenetic relatives. Therefore, it is proposed that strain NEAU-gq9(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora zeae sp. nov. is proposed. The type strain is NEAU-gq9(T) (=CGMCC 4.7092(T)=DSM 45882(T)).

  12. Nocardiopsis algeriensis sp. nov., an alkalitolerant actinomycete isolated from Saharan soil.

    PubMed

    Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Mathieu, Florence; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-02-01

    An alkalitolerant actinomycete strain, designated B32(T), was isolated from a Saharan soil sample collected from Adrar province (South of Algeria), and then investigated using a polyphasic taxonomic approach. The strain was observed to produce short chains of spores on the dichotomous branched aerial mycelium and formed a fragmented substrate mycelium. The optimum NaCl concentration for growth was found to be 0-5 % (w/v) and the optimum growth temperature and pH were found to be 25-35 °C and 7.0-10.0 °C, respectively. The diagnostic diamino acid in the cell-wall peptidoglycan was identified as meso-diaminopimelic acid. The predominant menaquinones of strain B32(T) were identified as MK-10 (H4) and MK-11 (H4). The major fatty acids were found to be iso-C16:0 and anteiso-C15:0. The diagnostic phospholipids detected were phosphatidylcholine, phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The chemotaxonomic properties of strain B32(T) are consistent with those shared by members of the genus Nocardiopsis. 16S rRNA gene sequence analysis indicated that strain B32(T) is most closely related to Nocardiopsis alba DSM 43377(T) (98.7 %), Nocardiopsis lucentensis DSM 44048(T) (98.6 %), Nocardiopsis aegyptia DSM 44442(T) (98.6 %), Nocardiopsis sinuspersici HM6(T) (98.6 %) and Nocardiopsis arvandica HM7(T) (98.5 %). However, the DNA-DNA relatedness values between strain B32(T) and the closely related type strains were 17.9, 14.6, 31.1, 27.1 and 14.1 %, respectively. Based on the combined genotypic and phenotypic evidence, it is proposed that strain B32(T) should be classified as representative of a novel species, for which the name Nocardiopsis algeriensis sp. nov. is proposed. The type strain is B32(T) (=DSM 45462(T) = CECT 8712(T)).

  13. Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-09-01

    A novel halophilic actinomycete strain, H23(T), was isolated from a Saharan soil sample collected in Djamâa (Oued Righ region), El-Oued province, South Algeria. Strain H23(T) was identified as a member of the genus Actinopolyspora by a polyphasic approach. Phylogenetic analysis showed that strain H23(T) had 16S rRNA gene sequence similarities ranging from 97.8 % (Actinopolyspora xinjiangensis TRM 40136(T)) to 94.8 % (Actinopolyspora mortivallis DSM 44261(T)). The strain grew optimally at pH 6.0-7.0, 28-32 °C and in the presence of 15-25 % (w/v) NaCl. The substrate mycelium was well developed and fragmented with age. The aerial mycelium produced long, straight or flexuous spore chains with non-motile, smooth-surfaced and rod-shaped spores. Strain H23(T) had MK-10 (H4) and MK-9 (H4) as the predominant menaquinones. The whole micro-organism hydrolysates mainly consisted of meso-diaminopimelic acid, galactose and arabinose. The diagnostic phospholipid detected was phosphatidylcholine. The major cellular fatty acids were anteiso-C17:0 (37.4 %), iso-C17:0 (14.8 %), iso-C15:0 (14.2 %), and iso-C16:0 (13.9 %). The genotypic and phenotypic data show that the strain represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora righensis sp. nov. is proposed, with the type strain H23(T) (=DSM 45501(T) = CCUG 63368(T) = MTCC 11562(T)).

  14. Actinopolyspora saharensis sp. nov., a novel halophilic actinomycete isolated from a Saharan soil of Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Mathieu, Florence; Lebrihi, Ahmed; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2013-04-01

    A novel halophilic actinomycete, strain H32(T), was isolated from a Saharan soil sample collected in El-Oued province, south Algeria. The isolate was characterized by means of polyphasic taxonomy. Optimal growth was determined to occur at 28-32 °C, pH 6.0-7.0 and in the presence of 15-25 % (w/v) NaCl. The strain was observed to produce abundant aerial mycelium, which formed long chains of rod-shaped spores at maturity, and fragmented substrate mycelium. The cell wall was determined to contain meso-diaminopimelic acid and the characteristic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-10(H4) and MK-9(H4). The predominant cellular fatty acids were determined to be anteiso C17:0, iso-C15:0 and iso-C16:0. The diagnostic phospholipid detected was phosphatidylcholine. Phylogenetic analyses based on the 16S rRNA gene sequence showed that this strain formed a distinct phyletic line within the radiation of the genus Actinopolyspora. The 16S rRNA gene sequence similarity indicated that strain H32(T) was most closely related to 'Actinopolyspora algeriensis' DSM 45476(T) (98.8 %) and Actinopolyspora halophila DSM 43834(T) (98.5 %). Furthermore, the result of DNA-DNA hybridization between strain H32(T) and the type strains 'A. algeriensis' DSM 45476(T), A. halophila DSM 43834(T) and Actinopolyspora mortivallis DSM 44261(T) demonstrated that this isolate represents a different genomic species in the genus Actinopolyspora. Moreover, the physiological and biochemical data allowed the differentiation of strain H32(T) from its closest phylogenetic neighbours. Therefore, it is proposed that strain H32(T) represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora saharensis sp. nov. is proposed. The type strain is H32(T) (=DSM 45459(T)=CCUG 62966(T)).

  15. Streptomonospora algeriensis sp. nov., a halophilic actinomycete isolated from soil in Algeria.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Riba, Amar; Zitouni, Abdelghani; Mathieu, Florence; Rohde, Manfred; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2014-08-01

    A halophilic actinomycete strain, designated H27(T), was isolated from a soil sample collected from a hypersaline habitat in Djelfa Province (North-Central Algeria), and then investigated using a polyphasic taxonomic approach. The strain was observed to produce poor aerial mycelium, which formed short chains of oval to cylindrical-shaped spores at maturity, and non fragmented substrate mycelium. The optimum NaCl concentration for growth was found to be 10-15 % (w/v) and the optimum growth temperature and pH were found to be 28-37 °C and 6-7, respectively. The diagnostic diamino acid in the cell-wall peptidoglycan was identified as meso-diaminopimelic acid. The predominant menaquinones of strain H27(T) were identified as MK-11 (H4) and MK-10 (H6). The major fatty acids were found to be iso-C16:0, anteiso-C17:0, 10 methyl C17:0 and 10 methyl C16:0. The diagnostic phospholipids detected were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol. The chemotaxonomic properties of strain H27(T) are consistent with those shared by members of the genus Streptomonospora. 16S rRNA gene sequence analysis indicated that strain H27(T) is most closely related to Streptomonospora alba DSM 44588(T) (98.8 %) and Streptomonospora flavalba DSM 45155(T) (98.7 %) whereas the DNA-DNA relatedness values between strain H27(T) and the two type strains were 17.1 and 57.9 %, respectively. Based on the combined genotypic and phenotypic evidence, it is proposed that strain H27(T) should be classified as representative of a novel species, for which the name Streptomonospora algeriensis sp. nov. is proposed. The type strain is H27(T) (=DSM 45604(T) =CCUG 63369(T) =MTCC 11563(T)).

  16. Saccharopolyspora ghardaiensis sp. nov., an extremely halophilic actinomycete isolated from Algerian Saharan soil.

    PubMed

    Meklat, Atika; Bouras, Noureddine; Zitouni, Abdelghani; Sabaou, Nasserdine; Mathieu, Florence; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter

    2014-04-01

    A novel halophilic actinomycete, strain designated H53(T), was isolated from a Saharan soil sample collected from Chaâbet Ntissa, Béni-isguen, Ghardaïa (South of Algeria) and was characterized taxonomically by means of polyphasic approach. Optimal growth was found to occur at 30-35 °C, pH 6-7 and in the presence of 15-25% (w/v) NaCl. The strain was observed to produce abundant aerial mycelium, which formed long chains of rod-shaped spores at maturity, and well developed and fragmented substrate mycelium. The cell wall was determined to contain meso-diaminopimelic acid; the diagnostic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-9(H₄) and MK-9(H₆). The predominant cellular fatty acids were determined to be iso- and anteiso-C17:0, iso-C15:0, and cis9 iso-C17:1. The diagnostic phospholipid detected was phosphatidylcholine. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Saccharopolyspora. Phylogenetic analyses on the basis of the 16S ribosomal RNA (rRNA) gene sequence showed that this strain formed a distinct phyletic line within the radiation of the genus Saccharopolyspora. The 16S rRNA sequence similarities between strain H53(T) and other members of the genus Saccharopolyspora ranged from 92.1 to 94.3%. The DNA G+C content of strain H53(T) was 72.6%. The genotypic and phenotypic data showed that the strain H53(T) represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora ghardaiensis sp. nov. is proposed, with the type strain H53(T) (=DSM 45606(T)=CCUG 63370(T)=CECT 8304(T)).

  17. Actinokineospora mzabensis sp. nov., a novel actinomycete isolated from Saharan soil.

    PubMed

    Aouiche, Adel; Bouras, Noureddine; Mokrane, Salim; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Sabaou, Nasserdine; Klenk, Hans-Peter

    2015-01-01

    A novel actinomycete strain, designated PAL84, was isolated from a Saharan soil sample collected from Béni-Isguen, Ghardaïa (South of Algeria). This strain was studied for its taxonomic position using a polyphasic approach and was identified as a member of the genus Actinokineospora. Phylogenetic analysis showed that strain PAL84 had 16S rRNA gene sequence similarities with members of the genus Actinokineospora ranging from 96.2 % (Actinokineospora inagensis DSM 44258(T)) to 97.8 % (Actinokineospora baliensis NBRC 104211(T)). The strain was observed to produce pinkish-purple aerial mycelium and purplish red substrate mycelium, which fragmented readily into chains of non-motile elements. The optimum growth temperature and pH were found to be 25-30 °C and 5.0-7.0, respectively. The cell-wall hydrolysate of strain PAL84 was found to contain meso-diaminopimelic acid and the diagnostic whole-cell sugars were identified as arabinose and galactose. The predominant menaquinone was identified as MK-9 (H4). The major fatty acids were found to be iso-C16:0, iso-C15:0, iso-C16:1 H and iso-C16:0 2OH. The diagnostic phospholipid detected was phosphatidylethanolamine. The genotypic and phenotypic data show that the strain represents a novel species of the genus Actinokineospora, for which the name Actinokineospora mzabensis sp. nov. is proposed, with the type strain PAL84(T) (=DSM 45961(T) = CECT 8578(T)).

  18. Nonomuraea flavida sp. nov., a novel species of soil actinomycete isolated from Aconitum napellus rhizosphere.

    PubMed

    Chen, Shaofeng; Shi, Jindi; Li, Dan; Wu, Yingying; Huang, Yaojian

    2015-11-01

    A novel actinomycete strain, YN-5-1T, isolated from the rhizosphere soil of a medicinal plant, Aconitum napellus, was characterized by a polyphasic approach to determine its taxonomic position. The strain showed highest 16S rRNA gene sequence similarities of 97.3, 97.2 and 97.1 % to Nonomuraea turkmeniaca DSM 43926T, Nonomuraea ferruginea DSM 43553T and Nonomuraea candida DSM 45086T, respectively. A wide range of genotypic and phenotypic characteristics, as well as levels of DNA-DNA relatedness between strain YN-5-1T and N. turkmeniaca DSM 43926T (57.46 %), N. ferruginea DSM 43553T (53.50 %) and N. candida DSM 45086T (48.80 %), distinguished the novel isolate from its closest phylogenetic neighbours. The morphological characteristics of strain YN-5-1T were typical of the genus Nonomuraea. Chemotaxonomic characteristics, such as diagnostic diamino acid of the peptidoglycan, whole-cell sugars, phospholipid type, major menaquinone and major fatty acids, further supported the assignment of strain YN-5-1T to the genus Nonomuraea. The G+C content of the genomic DNA was 72.1 mol%. Based on the above data, strain YN-5-1T is considered to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea flavida sp. nov. is proposed. The type strain is YN-5-1T ( = CCTCC AB 2012909T = KCTC 29143T).

  19. Saccharopolyspora griseoalba sp. nov., a novel actinomycete isolated from the Dead Sea.

    PubMed

    Jiang, Yingying; Wei, Xiaomin; Chen, Xiu; Jiang, Yi; Xue, Quanhong; Lai, Hangxian; Jiang, Chenglin

    2016-12-01

    A novel halotolerant actinomycete, designated strain AFM 10238(T), was isolated from a sediment sample collected from the Dead Sea of Israel. The isolate grew at 15-45 °C, pH 6-12 and with 0-15 % (w/v) NaCl. Strain AFM 10238(T) contains meso-diaminopimelic acid as cell wall diamino acid, and galactose and arabinose as the whole cell sugars. The major polar lipids are phosphatidylcholine, phosphatidylglycerol, and diphosphatidylglycerol. Major fatty acids are iso-C16:0, iso-C17:0, iso-C15:0, anteiso-C17:0 and C17:1 ω8c. MK-9(H4) is the predominant menaquinone and the DNA G + C content is 72.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM10238(T) belongs to the genus Saccharopolyspora. The 16S rRNA gene sequence similarity between strain AFM 10238(T) and its close neighbours, Saccharopolyspora halophila YIM 90500(T) , Saccharopolyspora spinosa DSM 44228(T), Saccharopolyspora dendranthemae KLBMP 1305(T) and Saccharopolyspora cebuensis DSM 45019(T) were 98.2, 97.2, 97.1 and 97.0 %, respectively. Sequence similarities to other type strains of this genus were below 97 %. DNA-DNA relatedness data, together with phenotypic and chemotaxonomic differences, clearly distinguished the isolate from its close neighbours. On the basis of the data from this polyphasic analysis, a novel species Saccharopolyspora griseoalba sp. nov. is proposed. The type strain is AFM 10238(T) (= DSM 46,663 = CGMCC 4.7124).

  20. Amycolatopsis flava sp. nov., a halophilic actinomycete isolated from Dead Sea.

    PubMed

    Wei, Xiaomin; Jiang, Yingying; Chen, Xiu; Jiang, Yi; Lai, Hangxian

    2015-10-01

    A novel halophilic, filamentous actinomycete, designated strain AFM 10111(T), was isolated from a sediment sample collected from the Dead Sea of Israel and its taxonomic position was established by using a polyphasic taxonomic approach. The isolate grew at 20-35 °C, pH 5-12 and with 1-30 % NaCl. The substrate mycelium is white or yellow, well developed, branched and fragments into squarish, rod-like elements. The isolate contained meso-diaminopimelic acid as cell-wall diamino acid, and arabinose and galactose as whole-cell sugars. The major menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine phosphatidylmethylethanolamine and one unidentified phospholipid. Major fatty acids were iso-C16:0, iso-C16:1 H, C17:1 ω6c. The DNA G + C content was 67.7 mol %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM 10111(T) belongs to the genus Amycolatopsis, and formed a distinct clade with Amycolatopsis marina CGMCC 4.3568(T) and Amycolatopsis palatopharyngis CGMCC 4.1729(T), with the sequence similarity 98.4 and 98.6 %. The level of DNA-DNA relatedness between the strain AFM 10111(T) and A. marina CGMCC 4.3568(T) and A. palatopharyngis CGMCC 4.1729(T) were 46.9 ± 3.08 and 49.4 ± 1.25 %. The combined genotypic and phenotypic data indicate that strain AFM 10111(T) represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis flava sp. nov. is proposed. The type strain is AFM 10111(T) (= DSM 46658(T) = CGMCC 4.7123(T)).

  1. Actinoallomurus bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Wang, Haiyan; Jin, Pinjiao; Zheng, Weijia; Chu, Liyang; Liu, Chongxi; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2015-08-01

    A novel endophytic actinomycete, strain NEAU-TX1-15(T), was isolated from moss, collected from Wuchang, Heilongjiang province, north China. A polyphasic taxonomic study was carried out to establish the status of strain NEAU-TX1-15(T). Morphological and chemotaxonomic properties of strain NEAU-TX1-15(T) are consistent with the description of the genus Actinoallomurus. Strain NEAU-TX1-15(T) was observed to form short spiral or looped spore chains on aerial hyphae. The cell wall peptidoglycan was found to contain lysine and meso-diaminopimelic acid. The major menaquinones were identified as MK-9(H6) and MK-9(H8). The only phospholipid identified was phosphatidylglycerol. The major fatty acid was identified as iso-C16:0. Analysis of the 16S rRNA gene sequence supports the assignment of the novel strain to the genus Actinoallomurus, as it exhibits 99.2 % gene sequence similarity to that of Actinoallomurus yoronensis NBRC 103686(T). However, the low level of DNA-DNA relatedness allowed the strain to be differentiated from its close relative. Moreover, strain NEAU-TX1-15(T) could also be differentiated from A. yoronensis NBRC 103686(T) and other Actinoallomurus species showing high 16S rRNA gene sequence similarity (>98.0 %) by cultural and physiological characteristics. Therefore, the combination of phenotypic and chemotaxonomic data, and the DNA-DNA hybridization value, indicated that strain NEAU-TX1-15(T) represents a novel species of the genus Actinoallomurus for which the name Actinoallomurus bryophytorum sp. nov. is proposed. The type strain is NEAU-TX1-15(T) (=CGMCC 4.7200(T) = JCM 30340(T)).

  2. Actinopolyspora biskrensis sp. nov., a novel halophilic actinomycete isolated from Northern Sahara.

    PubMed

    Saker, Rafika; Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2015-03-01

    A novel halophilic, filamentous actinomycete, designated H254(T), was isolated from a Saharan soil sample collected from Biskra (Northern Sahara), and subjected to a polyphasic taxonomic characterization. The strain is Gram-positive, aerobic, and halophilic, and the optimum NaCl concentration for growth is 15-20 % (w/v). The cell-wall hydrolysate contained meso-diaminopimelic acid, and the diagnostic whole-cell sugars were arabinose and galactose. The diagnostic phospholipid detected was phosphatidylcholine, and MK-9(H4) was the predominant menaquinone. The major fatty acid profiles were anteiso-C17:0 (32.8 %), C15:0 (28 %), and iso-C17:0 (12.3 %). Comparative analysis of the 16S rRNA gene sequences revealed that the strain H254(T) formed a well-separated sub-branch within the radiation of the genus Actinopolyspora, and the microorganism was most closely related to Actinopolyspora saharensis DSM 45459(T) (99.2 %), Actinopolyspora halophila DSM 43834(T) (99.1 %), and Actinopolyspora algeriensis DSM 45476(T) (99.0 %). Nevertheless, the strain had relatively lower mean values for DNA-DNA relatedness with the above strains (57.2, 68.4, and 45.6 %, respectively). Based on phenotypic features and phylogenetic position, we propose that strain H254(T) represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora biskrensis sp. nov. is proposed. The type strain of A. biskrensis is strain H254(T) (=DSM 46684(T) =CECT 8576(T)).

  3. Plantactinosporasoyae sp. nov., an endophytic actinomycete isolated from soybean root [Glycine max (L.) Merr].

    PubMed

    Guo, Xiaowei; Guan, Xuejiao; Liu, Chongxi; Jia, Feiyu; Li, Jiansong; Li, Jinmeng; Jin, Pinjiao; Li, Wenchao; Wang, Xiangjing; Xiang, Wensheng

    2016-07-01

    A novel actinomycete, designated strain NEAU-gxj3T, was isolated from soybean root [Glycine max (L.) Merr.] collected from Harbin, Heilongjiang Province, China, and characterized using a polyphasic approach. The 16S rRNA gene sequence of strain NEAU-gxj3T showed highest similarity to those of Micromonospora equina Y22T (98.2 %) and Plantactinospora endophytica YIM 68255T (98.0 %). Phylogenetic analysis based on the 16S rRNA gene and gyrB gene demonstrated that the isolate clustered with the members of the genus Plantactinospora. The chemotaxonomic properties of strain NEAU-gxj3Twere also consistent with those of members of the genus Plantactinospora. The cell wall contained meso-diaminopimelic acid and whole-cell sugars were xylose, glucose and galactose. The predominant menaquinones were MK-10(H6), MK-9(H8), MK-10(H2) and MK-10(H4). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were identified as anteiso-C17 : 0, iso-C16 : 0, iso-C15 : 0 and C15 : 0. A combination of DNA-DNA hybridization result and some phenotypic characteristics indicated that strain NEAU-gxj3Tcould be differentiated clearly from its closest phylogenetic relatives. Therefore, the strain is concluded to represent a novel species of the genus Plantactinospora, for which the name Plantactinospora soyae sp. nov. is proposed. The type strain is NEAU-gxj3T (=CGMCC 4.7221T=DSM 46832T).

  4. Sphaerisporangium dianthi sp. nov., an endophytic actinomycete isolated from a root of Dianthus chinensis L.

    PubMed

    Xing, Jia; Liu, Chongxi; Zhang, Yuejing; He, Hairong; Zhou, Ying; Li, Lianjie; Zhao, Junwei; Liu, Shuanghe; Wang, Xiangjing; Xiang, Wensheng

    2015-01-01

    A novel actinomycete, designated strain NEAU-CY18(T), was isolated from the root of a Chinese medicinal plant Dianthus chinensis L and subjected to a polyphasic taxonomic study. The novel strain was found to develop spherical sporangia with non-motile spores on aerial mycelium. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as madurose, mannose, ribose, galactose and glucose. The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified phospholipid. The predominant menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H6). The major fatty acids were identified as C17:0 10-methyl, iso-C16:0 and C16:0. EzTaxon-e analysis of the 16S rRNA gene sequence indicated that the strain belongs to the genus Sphaerisporangium and was most closely related to Sphaerisporangium cinnabarinum JCM 3291(T) (98.9 %) and Sphaerisporangium melleum JCM 13064(T) (98.3 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-CY18(T) forms a monophyletic clade with S. cinnabarinum JCM 3291(T), an association that was supported by a bootstrap value of 97 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. Comparisons of some phenotypic properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. cinnabarinum JCM 3291(T) and S. melleum JCM 13064(T). Therefore, it is concluded that strain NEAU-CY18(T) represents a novel Sphaerisporangium species, for which the name Sphaerisporangium dianthi sp. nov. is proposed. The type strain is NEAU-CY18(T) ( = CGMCC 4.7132(T) = DSM 46736(T)).

  5. 3-Methoxy-2-methyl-carbazole-1,4-quinone, carbazomycins D and F from Streptomyces sp. CMU-JT005.

    PubMed

    Ruanpanun, Pornthip; Dame, Zerihun Teklemariam; Laatsch, Hartmut; Lumyong, Saisamorn

    2011-09-01

    3-Methoxy-2-methyl-carbazole-1,4-quinone (1) together with carbazomycins D (2) and F (3) were isolated from the crude extract of Streptomyces CMU-JT005, an actinomycete with nematicidal activity. 3-Methoxy-2-methyl-carbazole-1,4-quinone is reported here for the first time from nature. In this paper, we describe the isolation and structure elucidation of the compounds together with the characterization of the Streptomyces strain CMU-JT005.

  6. Biosynthesis of silver nanoparticles by a new strain of Streptomyces sp. compared with Aspergillus fumigatus.

    PubMed

    Alani, Faiez; Moo-Young, Murray; Anderson, William

    2012-03-01

    Locally isolated strains of a thermoalkalotolerant Streptomyces sp. and Aspergillus fumigatus were used for the in vitro biosynthesis of silver nanoparticles from AgNO(3) solutions. An autolysed cell-free culture filtrate from each strain was used, indicating that the formation mechanism depends on intra-cellular components for both organisms, since culture broths had no significant nanoparticle formation potential. Nanoparticle formation was indicated by a change of the solution from colourless or light brown to dark brown after 24 h or more, and UV-visible spectroscopy and x-ray diffraction analysis confirmed the formation by both organisms. The initial formation kinetics were faster with Aspergillus, but formation continued for a longer period with Streptomyces, resulting in higher concentrations after 48 h. Transmission electron microscope images revealed well dispersed nanoparticles with diameters ranging from 15 to 45 nm from A. fumigatus, while those from Streptomyces sp. had a narrower size distribution of 15-25 nm. The higher productivity and preferred narrower size distribution of Streptomyces, together with its well established industrial use, may make it the preferred choice for further optimization studies.

  7. Isolation, Screening, and Identification of Novel Isolates of Actinomycetes from India for Antimicrobial Applications.

    PubMed

    Singh, Vineeta; Haque, Shafiul; Singh, Harshita; Verma, Jyoti; Vibha, Kumari; Singh, Rajbir; Jawed, Arshad; Tripathi, C K M

    2016-01-01

    The search for novel bioactive compounds from the natural environment has rapidly been gaining momentum with the increase in multi-drug resistant (MDR) pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569, and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes.

  8. Isolation, Screening, and Identification of Novel Isolates of Actinomycetes from India for Antimicrobial Applications

    PubMed Central

    Singh, Vineeta; Haque, Shafiul; Singh, Harshita; Verma, Jyoti; Vibha, Kumari; Singh, Rajbir; Jawed, Arshad; Tripathi, C. K. M.

    2016-01-01

    The search for novel bioactive compounds from the natural environment has rapidly been gaining momentum with the increase in multi-drug resistant (MDR) pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569, and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes. PMID:27999566

  9. Antimicrobial compounds from endophytic Streptomyces sp. BCC72023 isolated from rice (Oryza sativa L.).

    PubMed

    Supong, Khomsan; Thawai, Chitti; Choowong, Wilunda; Kittiwongwattana, Chokchai; Thanaboripat, Dusanee; Laosinwattana, Chamroon; Koohakan, Prommart; Parinthawong, Nonglak; Pittayakhajonwut, Pattama

    2016-05-01

    An endophytic actinomycete strain BCC72023 was isolated from rice (Oryza sativa L.) and identified as the genus Streptomyces, based on phenotypic, chemotaxonomic and 16S rRNA gene sequence analyses. The strain showed 99.80% similarity compared with Streptomyces samsunensis M1463(T). Chemical investigation led to the isolation of three macrolides, efomycins M (1), G (2) and oxohygrolidin (3), along with two polyethers, abierixin (4) and 29-O-methylabierixin (5). To our knowledge, this is the first report of efomycin M being isolated from a natural source. The compounds were identified using spectroscopic techniques and comparison with previously published data. All compounds exhibited antimalarial activity against the Plasmodium falciparum, K-1 strain, a multidrug-resistant strain, with IC50 values in a range of 1.40-5.23 μg/ml. In addition, these compounds were evaluated for biological activity against Mycobacterium tuberculosis, Bacillus cereus, Colletotrichum gloeosporioides and Colletotrichum capsici, as well as cytotoxicity against both cancerous (MCF-7, KB, NCI-H187) and non-cancerous (Vero) cells.

  10. Detoxification of Atrazine by Endophytic Streptomyces sp. Isolated from Sugarcane and Detection of Nontoxic Metabolite.

    PubMed

    Mesquini, Josiane A; Sawaya, Alexandra C H F; López, Begonã G C; Oliveira, Valéria M; Miyasaka, Natalia R S

    2015-12-01

    Atrazine is still one of the most used agricultural pesticides worldwide and it has been recognized as a major contaminant of surface and ground water. The aims of this research were to isolate an endophytic microorganism from leaves of sugarcane, evaluate its ability to degrade atrazine, and investigate the formation of metabolites. By sequencing of the 16S rRNA gene, the endophytic isolate atz2 was identified as Streptomyces sp. The reduction in atrazine concentration by Streptomyces sp. atz2 was 98 % and UHPLC-MS/MS analyses showed the appearance of an unknown metabolite observed as m/z 311. Ecotoxicity tests with an aquatic organism, Daphnia similis, confirmed that this metabolite was nontoxic. This mechanism of detoxification of atrazine is different from the ones of other free-living microorganisms that inhabit the soil or rhizosphere. The results show new aspects of atrazine detoxification, highlighting a new role of endophytic bacteria in plants.

  11. Nocardiopsis oceani sp. nov. and Nocardiopsis nanhaiensis sp. nov., actinomycetes isolated from marine sediment of the South China Sea.

    PubMed

    Pan, Hua-Qi; Zhang, Dao-Feng; Li, Li; Jiang, Zhao; Cheng, Juan; Zhang, Yong-Guang; Wang, Hong-Fei; Hu, Jiang-Chun; Li, Wen-Jun

    2015-10-01

    Two actinomycete strains, designated 10A08AT and 10A08BT, were isolated from marine sediment samples of the South China Sea and their taxonomic positions were determined by a polyphasic approach. The two Gram-stain-positive, aerobic strains produced branched substrate mycelium and aerial hyphae, and no diffusible pigment was produced in the media tested. At maturity, spore chains were formed on aerial hyphae and all mycelium fragmented with age. Whole-cell hydrolysates of both strains contained meso-diaminopimelic acid and no diagnostic sugars. Their predominant menaquinones (>10 %) were MK-9(H4), MK-9(H6) and MK-10(H6) for strain 10A08AT and MK-9(H4), MK-9(H6), MK-10(H4) and MK-10(H6) for strain 10A08BT. The polar lipids detected from the two strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and unknown phosphoglycolipids and phospholipids. The major fatty acids (>10 %) of both strains were iso-C16 : 0 and summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B). The genomic DNA G+C contents of strains 10A08AT and 10A08BT were 70.9 and 71.6 mol%, respectively. On the basis of 16S rRNA gene sequence similarities, the two strains were shown to be most closely related to species of the genus Nocardiopsis. DNA–DNA hybridization relatedness values of < 70 % between these two isolates and their closest neighbour, Nocardiopsis terrae YIM 90022T, and between the two strains supported the conclusion that they represent two novel species. Based on phylogenetic analysis and phenotypic and genotypic data, it is concluded that the two isolates belong to the genus Nocardiopsis, and the names Nocardiopsis oceani sp. nov. (type strain 10A08AT = DSM 45931T = BCRC 16951T) and Nocardiopsis nanhaiensis sp. nov. (type strain 10A08BT = CGMCC 47227T = BCRC 16952T) are proposed.

  12. Investigation on antimicrobial agents of the terrestrial Streptomyces sp. BCC71188.

    PubMed

    Supong, Khomsan; Sripreechasak, Paranee; Tanasupawat, Somboon; Danwisetkanjana, Kannawat; Rachtawee, Pranee; Pittayakhajonwut, Pattama

    2017-01-01

    The terrestrial actinomycete strain BCC71188 was identified as Streptomyces by its morphology (having spiral chain spore on the aerial mycelium), chemotaxonomy (containing LL-diaminopimelic acid in the cell wall), and 16S rRNA gene sequence analysis [showing high similarity values compared with Streptomyces samsunensis M1463(T) (99.85 %) and Streptomyces malaysiensis NBRC 16446(T) (99.40 %)]. The crude extract exhibited antimalarial against Plasmodium falciparum (IC50 0.19 μg/ml), anti-TB against Mycobacterial tuberculosis (MIC 6.25 μg/ml), and antibacterial against Bacillus cereus (MIC 1.56 μg/ml) activities. Therefore, chemical investigation was conducted by employing bioassay-guided method and led to the isolation of 19 compounds including two cyclic peptides (1-2), five macrolides (3-7), new naphthoquinone (8), nahuoic acid C (9), geldanamycin derivatives (10-13), cyclooctatin (14), germicidins A (15) and C (16), actinoramide A (17), abierixin, and 29-O-methylabierixin. These isolated compounds were evaluated for antimicrobial activity, such as antimalarial, anti-TB, and antibacterial activities, and for cytotoxicity against both cancerous (MCF-7, KB, NCI-H187) and non-cancerous (Vero) cells. Compounds 1-7, 10-14 exhibited antimalarial (IC50 0.22-7.14 μg/ml), and elaiophylin analogs (4-6) displayed anti-TB (MIC 0.78-12.00 μg/ml) and B. cereus (MIC 0.78-3.13 μg/ml) activities. Compounds 1, 2, 14, and abierixin displayed weak cytotoxicity, indicating a potential for antimicrobial agents.

  13. Broad Spectrum Antimicrobial Activity of Forest-Derived Soil Actinomycete, Nocardia sp. PB-52

    PubMed Central

    Sharma, Priyanka; Kalita, Mohan C.; Thakur, Debajit

    2016-01-01

    A mesophilic actinomycete strain designated as PB-52 was isolated from soil samples of Pobitora Wildlife Sanctuary of Assam, India. Based on phenotypic and molecular characteristics, the strain was identified as Nocardia sp. which shares 99.7% sequence similarity with Nocardia niigatensis IFM 0330 (NR_112195). The strain is a Gram-positive filamentous bacterium with rugose spore surface which exhibited a wide range of antimicrobial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA), Gram-negative bacteria, and yeasts. Optimization for the growth and antimicrobial activity of the strain PB-52 was carried out in batch culture under shaking condition. The optimum growth and antimicrobial potential of the strain were recorded in GLM medium at 28°C, initial pH 7.4 of the medium and incubation period of 8 days. Based on polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS) gene-targeted PCR amplification, the occurrence of both of these biosynthetic pathways was detected which might be involved in the production of antimicrobial compounds in PB-52. Extract of the fermented broth culture of PB-52 was prepared with organic solvent extraction method using ethyl acetate. The ethyl acetate extract of PB-52 (EA-PB-52) showed lowest minimum inhibitory concentration (MIC) against S. aureus MTCC 96 (0.975 μg/mL) whereas highest was recorded against Klebsiella pneumoniae ATCC 13883 (62.5 μg/mL). Scanning electron microscopy (SEM) revealed that treatment of the test microorganisms with EA-PB-52 destroyed the targeted cells with prominent loss of cell shape and integrity. In order to determine the constituents responsible for its antimicrobial activity, EA-PB-52 was subjected to chemical analysis using gas chromatography-mass spectrometry (GC-MS). GC-MS analysis showed the presence of twelve different chemical constituents in the extract, some of which are reported to possess diverse biological activity. These

  14. Exploring the diversity and metabolic potential of actinomycetes from temperate marine sediments from Newfoundland, Canada.

    PubMed

    Duncan, K R; Haltli, B; Gill, K A; Correa, H; Berrué, F; Kerr, R G

    2015-01-01

    Marine sediments from Newfoundland, Canada were explored for biotechnologically promising Actinobacteria using culture-independent and culture-dependent approaches. Culture-independent pyrosequencing analyses uncovered significant actinobacterial diversity (H'-2.45 to 3.76), although the taxonomic diversity of biotechnologically important actinomycetes could not be fully elucidated due to limited sampling depth. Assessment of culturable actinomycete diversity resulted in the isolation of 360 actinomycetes representing 59 operational taxonomic units, the majority of which (94 %) were Streptomyces. The biotechnological potential of actinomycetes from NL sediments was assessed by bioactivity and metabolomics-based screening of 32 representative isolates. Bioactivity was exhibited by 41 % of isolates, while 11 % exhibited unique chemical signatures in metabolomics screening. Chemical analysis of two isolates resulted in the isolation of the cytotoxic metabolite 1-isopentadecanoyl-3β-D-glucopyranosyl-X-glycerol from Actinoalloteichus sp. 2L868 and sungsanpin from Streptomyces sp. 8LB7. These results demonstrate the potential for the discovery of novel bioactive metabolites from actinomycetes isolated from Atlantic Canadian marine sediments.

  15. Streptomyces caldifontis sp. nov., isolated from a hot water spring of Tatta Pani, Kotli, Pakistan.

    PubMed

    Amin, Arshia; Ahmed, Iftikhar; Khalid, Nauman; Osman, Ghenijan; Khan, Inam Ullah; Xiao, Min; Li, Wen-Jun

    2017-01-01

    A Gram-staining positive, non-motile, rod-shaped, catalase positive and oxidase negative bacterium, designated NCCP-1331(T), was isolated from a hot water spring soil collected from Tatta Pani, Kotli, Azad Jammu and Kashmir, Pakistan. The isolate grew at a temperature range of 18-40 °C (optimum 30 °C), pH 6.0-9.0 (optimum 7.0) and with 0-6 % NaCl (optimum 2 % NaCl (w/v)). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain NCCP-1331(T) belonged to the genus Streptomyces and is closely related to Streptomyces brevispora BK160(T) with 97.9 % nucleotide similarity, followed by Streptomyces drosdowiczii NRRL B-24297(T) with 97.8 % nucleotide similarity. The DNA-DNA relatedness values of strain NCCP-1331(T) with S. brevispora KACC 21093(T) and S. drosdowiczii CBMAI 0498(T) were 42.7 and 34.7 %, respectively. LL-DAP was detected as diagnostic amino acid along with alanine, glycine, leucine and glutamic acid. The isolate contained MK-9(H8) as the predominant menaquinone. Major polar lipids detected in NCCP-1331(T) were phosphatidylethanolamine, phosphatidylinositol and unidentified phospholipids. Major fatty acids were iso-C16: 0, summed feature 8 (18:1 ω7c/18:1 ω6c), anteiso-C15:0 and C16:0. The genomic DNA G + C content was 69.8 mol %. On the basis of phylogenetic, phenotypic and chemotaxonomic analysis, it is concluded that strain NCCP-1331(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces caldifontis sp. nov. is proposed. The type strain is NCCP-1331(T) (=KCTC 39537(T) = CPCC 204147(T)).

  16. Streptomyces humi sp. nov., an actinobacterium isolated from soil of a mangrove forest.

    PubMed

    Zainal, Nurullhudda; Ser, Hooi-Leng; Yin, Wai-Fong; Tee, Kok-Keng; Lee, Learn-Han; Chan, Kok-Gan

    2016-03-01

    A novel Streptomyces strain, MUSC 119(T), was isolated from a soil collected from a mangrove forest. Cells of MUSC 119(T) stained Gram-positive and formed light brownish grey aerial mycelium and grayish yellowish brown substrate mycelium on ISP 2 medium. A polyphasic approach was used to determine the taxonomic status of strain MUSC 119(T), which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Streptomyces. The cell wall peptidoglycan consisted of LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9(H8), MK-9(H6) and MK-9(H4). The polar lipid profile consisted of phosphatidylinositol, phosphatidylethanolamine, glycolipids, diphosphatidylglycerol and four phospholipids. The predominant cellular fatty acids were anteiso-C15:0, iso-C16:0, and anteiso-C17:0. The cell wall sugars were glucose, mannose, ribose and rhamnose. The phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain MUSC119(T) to be closely related to Streptomyces rhizophilus JR-41(T) (99.0 % sequence similarity), S. panaciradicis 1MR-8(T) (98.9 %), S. gramineus JR-43(T) (98.8 %) and S. graminisoli JR-19(T) (98.7 %). These results suggest that MUSC 119(T) should be placed within the genus Streptomyces. DNA-DNA relatedness values between MUSC 119(T) to closely related strains ranged from 14.5 ± 1.3 to 27.5 ± 0.7 %. The G+C content was determined to be 72.6 mol %. The polyphasic study of MUSC 119(T) showed that this strain represents a novel species, for which the name Streptomyces humi sp. nov. is proposed. The type strain of S. humi is MUSC 119(T) (=DSM 42174(T) = MCCC 1K00505(T)).

  17. The Electrospray Ionization - Mass Spectra of Erythromycin A Obtained from a Marine Streptomyces sp. Mutant

    PubMed Central

    El-Bondkly, A. M.; Abd-Alla, Howaida I.; Shaaban, M.; Shaaban, K. A.

    2008-01-01

    In our ongoing search for production improvements of bioactive secondary metabolites from marine Streptomyces through the induction of mutations using UV light, out of 145 isolates, mutant 10/14 was able to produce potent antibacterial metabolites other than the parent strain as established by chromatographic analysis. Up-scaling fermentation of mutant 10/14, followed by working up and isolation delivered five metabolites, phenazine, 1-acetyl-β -carboline, perlolyrin and erythromycin A, along with an oily substance. The latter two compounds were responsible for the antibacterial activity of the strain. In this article, we discuss with the mutation of the marine Streptomyces sp. AH2, bioactivity evaluation, fermentation and isolation of the microbial metabolites. Moreover, we study to first time in detail the 1D and 2D NMR and ESI MS data including ESI MS2 and MS3 patterns combined with HRESI MS of erythromycin A. PMID:20046738

  18. Versatility of Streptomyces sp. M7 to bioremediate soils co-contaminated with Cr(VI) and lindane.

    PubMed

    Aparicio, JuanDaniel; Solá, María Zoleica Simón; Benimeli, Claudia Susana; Amoroso, María Julia; Polti, Marta Alejandra

    2015-06-01

    The aim of this work was to study the impact of environmental factors on the bioremediation of Cr(VI) and lindane contaminated soil, by an actinobacterium, Streptomyces sp. M7, in order to optimize the process. Soil samples were contaminated with 25 µg kg(-1) of lindane and 50 mg kg(-1) of Cr(VI) and inoculated with Streptomyces sp. M7. The lowest inoculum concentration which simultaneously produced highest removal of Cr(VI) and lindane was 1 g kg(-1). The influence of physical and chemical parameters was assessed using a full factorial design. The factors and levels tested were: Temperature: 25, 30, 35°C; Humidity: 10%, 20%, 30%; Initial Cr(VI) concentration: 20, 50, 80 mg kg(-1); Initial lindane concentration: 10, 25, 40 µg kg(-1). Streptomyces sp. M7 exhibited strong versatility, showing the ability to bioremediate co-contaminated soil samples at several physicochemical conditions. Streptomyces sp. M7 inoculum size was optimized. Also, it was fitted a model to study this process, and it was possible to predict the system performance, knowing the initial conditions. Moreover, optimum temperature and humidity conditions for the bioremediation of soil with different concentrations of Cr(VI) and lindane were determined. Lettuce seedlings were a suitable biomarker to evaluate the contaminants mixture toxicity. Streptomyces sp. M7 carried out a successful bioremediation, which was demonstrated through ecotoxicity test with Lactuca sativa.

  19. The variation of antioxidant defense system of Streptomyces sp. M4018 with respect to carbon sources.

    PubMed

    Kayalı, Hulya Ayar; Sazak, Anıl; Sahin, Nevzat; Tarhan, Leman

    2012-01-01

    The effect of glycerol, glucose, and starch as carbon sources on the antioxidant defense system such as superoxide dismutase (SOD) and catalase (CAT) activities, pyruvate levels, and membrane lipid peroxidation (LPO) levels of Streptomyces sp. M4018, after isolation from the rhizosphere samples of Colutea arborescens and identification as a strain of S. hiroshimensis based on phenotypic and genotypic characteristics, were investigated. As an antioxidant defense enzyme, SOD activities increased up to 20 g/L of glycerol and 15 g/L of starch, while they showed negative correlation with glucose concentration. CAT activity variations of glycerol- and glucose-supplemented mediums showed significant positive correlations with the trend of SOD activities. However, CAT activity, in contrast to SOD, in Streptomyces sp. M4018 tended to decrease as the starch concentration increased. The production of pyruvate increased with respect to glycerol and starch up to 15 g/L, while it was positively correlated with glucose concentration. The highest pyruvate production was seen at 20 g/L glucose. Membrane LPO levels were negatively correlated with the activities of SOD and CAT enzymes, and the minimum LPO level was determined at 5 g/L of glucose, where SOD and CAT activities reached their maximum levels. Nevertheless, the higher SOD and CAT activities in a wider range of incubation period compared to the beginning by resulting in insignificant increases in membrane LPO levels showed the unusual antioxidant response capacities of the in Streptomyces sp. M4018 against the potentially deleterious effects of reactive oxygen species (ROS) for glycerol, glucose, and starch as carbon sources.

  20. Biodiversity of Actinomycetes associated with Caribbean sponges and their potential for natural product discovery.

    PubMed

    Vicente, Jan; Stewart, Allison; Song, Bongkeun; Hill, Russell T; Wright, Jeffrey L

    2013-08-01

    Marine actinomycetes provide a rich source of structurally unique and bioactive secondary metabolites. Numerous genera of marine actinomycetes have been isolated from marine sediments as well as several sponge species. In this study, 16 different species of Caribbean sponges were collected from four different locations in the coastal waters off Puerto Rico in order to examine diversity and bioactive metabolite production of marine actinomycetes in Caribbean sponges. Sediments were also collected from each location, in order to compare actinomycete communities between these two types of samples. A total of 180 actinomycetes were isolated and identified based on 16S rRNA gene analysis. Phylogenetic analysis revealed the presence of at least 14 new phylotypes belonging to the genera Micromonospora, Verruscosispora, Streptomyces, Salinospora, Solwaraspora, Microbacterium and Cellulosimicrobium. Seventy-eight of the isolates (19 from sediments and 59 from sponges) shared 100 % sequence identity with Micromonospora sp. R1. Despite having identical 16S rRNA sequences, the bioactivity of extracts and subsequent fractions generated from the fermentation of both sponge- and sediment-derived isolates identical to Micromonospora sp. R1 varied greatly, with a marked increase in antibiotic metabolite production in those isolates derived from sponges. These results indicate that the chemical profiles of isolates with high 16S rRNA sequence homology to known strains can be diverse and dependent on the source of isolation. In addition, seven previously reported dihydroquinones produced by five different Streptomyces strains have been purified and characterized from one Streptomyces sp. strain isolated in this study from the Caribbean sponge Agelas sceptrum.

  1. Partial characterization of cold active amylases and proteases of Streptomyces sp. from Antarctica

    PubMed Central

    Cotârleţ, Mihaela; Negoiţă, Teodor Gh.; Bahrim, Gabriela E.; Stougaard, Peter

    2011-01-01

    The aim of this study was to isolate novel enzyme-producing bacteria from vegetation samples from East Antarctica and also to characterize them genetically and biochemically in order to establish their phylogeny. The ability to grow at low temperature and to produce amylases and proteases cold-active was also tested. The results of the 16S rRNA gene sequence analysis showed that the 4 Alga rRNA was 100% identical to the sequences of Streptomyces sp. rRNA from Norway and from the Solomon Islands. The Streptomyces grew well in submerged system at 20°C, cells multiplication up to stationary phase being drastically increased after 120 h of submerged cultivation. The beta-amylase production reached a maximum peak after seven days, while alpha-amylase and proteases were performing biosynthesis after nine days of submerged cultivation at 20°C. Newly Streptomyces were able to produce amylase and proteases in a cold environment. The ability to adapt to low temperature of these enzymes could make them valuable ingredients for detergents, the food industry and bioremediation processes which require low temperatures. PMID:24031702

  2. Streptomyces antioxidans sp. nov., a Novel Mangrove Soil Actinobacterium with Antioxidative and Neuroprotective Potentials

    PubMed Central

    Ser, Hooi-Leng; Tan, Loh Teng-Hern; Palanisamy, Uma D.; Abd Malek, Sri N.; Yin, Wai-Fong; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2016-01-01

    A novel strain, Streptomyces antioxidans MUSC 164T was recovered from mangrove forest soil located at Tanjung Lumpur, Malaysia. The Gram-positive bacterium forms yellowish-white aerial and brilliant greenish yellow substrate mycelium on ISP 2 agar. A polyphasic approach was used to determine the taxonomy status of strain MUSC 164T. The strain showed a spectrum of phylogenetic and chemotaxonomic properties consistent with those of the members of the genus Streptomyces. The cell wall peptidoglycan was determined to contain LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9(H6) and MK-9(H8), while the identified polar lipids consisted of aminolipid, diphosphatidylglycerol, glycolipid, hydroxyphosphatidylethanolamine, phospholipid, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol and lipid. The cell wall sugars consist of galactose, glucose and ribose. The predominant cellular fatty acids (>10.0%) were identified as iso-C15:0 (34.8%) and anteiso-C15:0(14.0%). Phylogenetic analysis identified that closely related strains for MUSC 164T as Streptomyces javensis NBRC 100777T (99.6% sequence similarity), Streptomyces yogyakartensis NBRC 100779T (99.6%) and Streptomyces violaceusniger NBRC 13459T (99.6%). The DNA–DNA relatedness values between MUSC 164T and closely related type strains ranged from 23.8 ± 0.3% to 53.1 ± 4.3%. BOX-PCR fingerprints comparison showed that MUSC 164T exhibits a unique DNA profile, with DNA G + C content determined to be 71.6 mol%. Based on the polyphasic study of MUSC 164T, it is concluded that this strain represents a novel species, for which the name Streptomyces antioxidans sp. nov. is proposed. The type strain is MUSC 164T (=DSM 101523T = MCCC 1K01590T). The extract of MUSC 164T showed potent antioxidative and neuroprotective activities against hydrogen peroxide. The chemical analysis of the extract revealed that the strain produces pyrazines and phenolic-related compounds that could explain

  3. Natural occurrence of organofluorine and other constituents from Streptomyces sp. TC1.

    PubMed

    Jaivel, Nanjundan; Uvarani, Chokkalingam; Rajesh, Ramasamy; Velmurugan, Devadasan; Marimuthu, Ponnusamy

    2014-01-24

    Antioxidant-directed fractionation of an ethyl acetate extract of Streptomyces sp. TC1 resulted in the isolation of a novel secondary metabolite with an aromatic organofluorine scaffold (1), an atypical tripod-type triallyl phenol (2), and a leucine residue comprised polyamine (3). Their structures were established by comprehensive spectroscopic analysis of 1D and 2D NMR data, and compound 1 was confirmed by (19)F NMR and single-crystal X-ray diffraction studies. The absolute configuration of compound 3 was assigned by comparison of its ECD spectra and quantum chemical ECD calculations. Of these, compound 1 displayed antioxidant and DNA and protein binding properties.

  4. Structure of a putative fluorinated natural product from Streptomyces sp. TC1.

    PubMed

    Aldemir, Hülya; Kohlhepp, Stefanie V; Gulder, Tanja; Gulder, Tobias A M

    2014-11-26

    Fluorine-containing natural products are extremely rare. The recent report on the isolation and biological activity of the bacterial secondary metabolite 3-(3,5-di-tert-butyl-4-fluorophenyl)propionic acid was thus highly remarkable. The compound contained the first aromatic fluorine substituent known to date in any natural product. The promise to discover an enzyme capable of aromatic fluorination in the producing strain Streptomyces sp. TC1 prompted our immediate interest. A close inspection of the originally reported analytical data of the fluoro metabolite revealed inconsistencies that triggered us to validate the reported structure. The results of these efforts are presented in this communication.

  5. Leupeptazin, a highly modified tripeptide isolated from cultures of a Streptomyces sp. inhibits cathepsin K.

    PubMed

    Kruglyak, Natasha; Williams, David E; Chen, Henry; Law, Simon; Kaleta, Jadwiga; Villanueva, Ivan; Davies, Julian E; Andersen, Raymond J; Brömme, Dieter

    2017-03-15

    Using a human cathepsin K-targeting inhibitor screen, a new leupeptin analogue, leupeptazin (1), containing an unprecedented piperidinotriazine moiety, was isolated from a liquid culture of soil Streptomyces sp. IS2-4 collected in northern Italy. The structure of leupeptazin was established using HRESIMS as well as 1D and 2D NMR data. The inhibitory activity of the compound towards the collagenase cathepsin K was tested in vitro to reveal moderate activity with an inhibition constant, Ki, of 44μM.

  6. Isolation, identification, and cytotoxicity of a new isobenzofuran derivative from marine Streptomyces sp. W007

    NASA Astrophysics Data System (ADS)

    Zhang, Hongyu; Xie, Zeping; Lou, Tingting; Jiang, Peng

    2016-03-01

    A new isobenzofuran derivative ( 1) was isolated from the marine Streptomyces sp. W007 and its structure was determined through extensive spectroscopic analyses, including 1D-NMR, 2D-NMR, and ESI-MS. The absolute configuration of compound 1 was determined by a combination of experimental analyses and comparison with reported data, including biogenetic reasoning, J-coupling analysis, NOESY, and 1H-1HCOSY. Compound 1 exhibited no cytotoxicity against human cells of gastric cancer BGC-823, lung cancer A549, and breast cancer MCF7.

  7. New α-glucosidase inhibitors from marine algae-derived Streptomyces sp. OUCMDZ-3434

    PubMed Central

    Chen, Zhengbo; Hao, Jiejie; Wang, Liping; Wang, Yi; Kong, Fandong; Zhu, Weiming

    2016-01-01

    Wailupemycins H (1) and I (2) with a new skeleton coupled two 6-(2-phenylnaphthalene-1-yl)pyrane-2-one nuclei to a –CH2– linkage were identified from the culture of Streptomyces sp. OUCMDZ-3434 associated with the marine algae, Enteromorpha prolifera. Compounds 1 and 2 are two new α-glucosidase inhibitors with the Ki/IC50 values of 16.8/19.7 and 6.0/8.3 μM, respectively. In addition, the absolute configurations of wailupemycins D (3) and E (4) are also resolved in this paper for the first time. PMID:26822662

  8. A new diketopiperazine derivative from a deep sea-derived Streptomyces sp. SCSIO 04496.

    PubMed

    Luo, Minghe; Tang, Guiling; Ju, Jianhua; Lu, Laichun; Huang, Hongbo

    2016-01-01

    A new diketopiperazine (DKP) derivative, (6R,3Z)-3-benzylidene-6-isobutyl-1-methyl piperazine-2,5-dione (1), as well as five known DKPs 2-6 was isolated from a deep sea-derived Streptomyces sp. SCSIO 04496. The structure of 1 was elucidated using a combination of 1D and 2D NMR, HR-ESI-MS and chiral-phase HPLC techniques. Compounds 1-6 did not show cytotoxic activity at a concentration of 100 μM in bioactivity assay.

  9. Draft genome sequence of Streptomyces sp. strain F1, a potential source for glycoside hydrolases isolated from Brazilian soil.

    PubMed

    Melo, Ricardo Rodrigues de; Persinoti, Gabriela Felix; Paixão, Douglas Antonio Alvaredo; Squina, Fábio Márcio; Ruller, Roberto; Sato, Helia Harumi

    2017-02-10

    Here, we show the draft genome sequence of Streptomyces sp. F1, a strain isolated from soil with great potential for secretion of hydrolytic enzymes used to deconstruct cellulosic biomass. The draft genome assembly of Streptomyces sp. strain F1 has 69 contigs with a total genome size of 8,142,296bp and G+C 72.65%. Preliminary genome analysis identified 175 proteins as Carbohydrate-Active Enzymes, being 85 glycoside hydrolases organized in 33 distinct families. This draft genome information provides new insights on the key genes encoding hydrolytic enzymes involved in biomass deconstruction employed by soil bacteria.

  10. Streptomyces cameroonensis sp. nov., a Geldanamycin Producer That Promotes Theobroma cacao Growth.

    PubMed

    Boudjeko, Thaddée; Tchinda, Romaric Armel Mouafo; Zitouni, Mina; Nana, Joëlle Aimée Vera Tchatchou; Lerat, Sylvain; Beaulieu, Carole

    2017-03-31

    The taxonomy of an actinobacterial strain, designated JJY4(T), was established using a polyphasic approach. JJY4(T) was isolated from the rhizosphere of Chromolaena odorata in Yaoundé (Cameroon) during a project for the selection of biological control agents. Strain JJY4(T) exhibited antimicrobial activities against bacteria, fungi, and oomycetes. Strain JJY4(T) also exhibited the traits of plant growth-promoting rhizobacteria such as the solubilization of inorganic phosphate, production of siderophores and indole-3-acetic acid, and 1-aminocyclopropane-1-carboxylate deaminase activity. In planta assays performed on cocoa plantlets confirmed that strain JJY4(T) exhibited strong abilities to promote plant growth and protect against Phytophthora megakarya, the main causal agent of cocoa pod rot. The formation of rugose-ornamented spores in spiral spore chains by strain JJY4(T) is a typical feature of members found in the Streptomyces violaceusniger clade and, similar to some members of the clade, strain JJY4(T) produces geldanamycin. A phylogenetic analysis based on 16S rRNA gene sequences confirmed this classification and suggests that strain JJY4(T) be added to the subclade constituted of the type strains Streptomyces malaysiensis DSM 41697(T) and Streptomyces samsunensis DSM 42010(T). However, DNA-DNA relatedness and physiological characteristics allowed for the differentiation of strain JJY4(T) from its closest phylogenetic relatives. Based on these results, strain JJY4(T) (=NRRL B-65369, =NBRC 112705) appears to represent a novel species in the S. violaceusniger clade for which the proposed name is Streptomyces cameroonensis sp. nov.

  11. Streptomyces cameroonensis sp. nov., a Geldanamycin Producer That Promotes Theobroma cacao Growth

    PubMed Central

    Boudjeko, Thaddée; Tchinda, Romaric Armel Mouafo; Zitouni, Mina; Nana, Joëlle Aimée Vera Tchatchou; Lerat, Sylvain; Beaulieu, Carole

    2017-01-01

    The taxonomy of an actinobacterial strain, designated JJY4T, was established using a polyphasic approach. JJY4T was isolated from the rhizosphere of Chromolaena odorata in Yaoundé (Cameroon) during a project for the selection of biological control agents. Strain JJY4T exhibited antimicrobial activities against bacteria, fungi, and oomycetes. Strain JJY4T also exhibited the traits of plant growth-promoting rhizobacteria such as the solubilization of inorganic phosphate, production of siderophores and indole-3-acetic acid, and 1-aminocyclopropane-1-carboxylate deaminase activity. In planta assays performed on cocoa plantlets confirmed that strain JJY4T exhibited strong abilities to promote plant growth and protect against Phytophthora megakarya, the main causal agent of cocoa pod rot. The formation of rugose-ornamented spores in spiral spore chains by strain JJY4T is a typical feature of members found in the Streptomyces violaceusniger clade and, similar to some members of the clade, strain JJY4T produces geldanamycin. A phylogenetic analysis based on 16S rRNA gene sequences confirmed this classification and suggests that strain JJY4T be added to the subclade constituted of the type strains Streptomyces malaysiensis DSM 41697T and Streptomyces samsunensis DSM 42010T. However, DNA–DNA relatedness and physiological characteristics allowed for the differentiation of strain JJY4T from its closest phylogenetic relatives. Based on these results, strain JJY4T (=NRRL B-65369, =NBRC 112705) appears to represent a novel species in the S. violaceusniger clade for which the proposed name is Streptomyces cameroonensis sp. nov. PMID:28260703

  12. Actinomadurol, an Antibacterial Norditerpenoid from a Rare Actinomycete, Actinomadura sp. KC 191.

    PubMed

    Shin, Bora; Kim, Byung-Yong; Cho, Eunji; Oh, Ki-Bong; Shin, Jongheon; Goodfellow, Michael; Oh, Dong-Chan

    2016-07-22

    A new secondary metabolite, actinomadurol (1), was isolated along with the known compound JBIR-65 (2) from a rare actinomycete, Actinomadura strain KC 191. The structure of 1 was established as a rare member of the bacterial C-19 norditerpenoid class by NMR data and ECD calculations. The absolute configuration of 2, which was previously reported without stereochemical analysis, was determined by using the modified Mosher's method and ECD calculations. Actinomadurol (1) exhibited potent antibacterial activity against pathogenic strains, such as Staphylococcus aureus, Kocuria rhizophila, and Proteus hauseri (MIC = 0.39-0.78 μg/mL), whereas JBIR-65 (2) showed no antibacterial activity.

  13. Taxonomic evaluation of Streptomyces albus and related species using multilocus sequence analysis and proposals to emend the description of Streptomyces albus and describe Streptomyces pathocidini sp. nov

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In phylogenetic analyses of the genus Streptomyces using 16S rRNA gene sequences, Streptomyces albus subsp. albus NRRL B-1811T forms a cluster with 5 other species having identical or nearly identical 16S rRNA gene sequences. Moreover, the morphological and physiological characteristics of these oth...

  14. -Genomic data mining of the marine actinobacteria Streptomyces sp. H-KF8 unveils insights into multi-stress related genes and metabolic pathways involved in antimicrobial synthesis.

    PubMed

    Undabarrena, Agustina; Ugalde, Juan A; Seeger, Michael; Cámara, Beatriz

    2017-01-01

    Streptomyces sp. H-KF8 is an actinobacterial strain isolated from marine sediments of a Chilean Patagonian fjord. Morphological characterization together with antibacterial activity was assessed in various culture media, revealing a carbon-source dependent activity mainly against Gram-positive bacteria (S. aureus and L. monocytogenes). Genome mining of this antibacterial-producing bacterium revealed the presence of 26 biosynthetic gene clusters (BGCs) for secondary metabolites, where among them, 81% have low similarities with known BGCs. In addition, a genomic search in Streptomyces sp. H-KF8 unveiled the presence of a wide variety of genetic determinants related to heavy metal resistance (49 genes), oxidative stress (69 genes) and antibiotic resistance (97 genes). This study revealed that the marine-derived Streptomyces sp. H-KF8 bacterium has the capability to tolerate a diverse set of heavy metals such as copper, cobalt, mercury, chromate and nickel; as well as the highly toxic tellurite, a feature first time described for Streptomyces. In addition, Streptomyces sp. H-KF8 possesses a major resistance towards oxidative stress, in comparison to the soil reference strain Streptomyces violaceoruber A3(2). Moreover, Streptomyces sp. H-KF8 showed resistance to 88% of the antibiotics tested, indicating overall, a strong response to several abiotic stressors. The combination of these biological traits confirms the metabolic versatility of Streptomyces sp. H-KF8, a genetically well-prepared microorganism with the ability to confront the dynamics of the fjord-unique marine environment.

  15. Minimal Streptomyces sp. strain C5 daunorubicin polyketide biosynthesis genes required for aklanonic acid biosynthesis.

    PubMed Central

    Rajgarhia, V B; Strohl, W R

    1997-01-01

    The structure of the Streptomyces sp. strain C5 daunorubicin type II polyketide synthase (PKS) gene region is different from that of other known type II PKS gene clusters. Directly downstream of the genes encoding ketoacylsynthase alpha and beta (KS alpha, KS beta) are two genes (dpsC, dpsD) encoding proteins of unproven function, both absent from other type II PKS gene clusters. Also in contrast to other type II PKS clusters, the gene encoding the acyl carrier protein (ACP), dpsG, is located about 6.8 kbp upstream of the genes encoding the daunorubicin KS alpha and KS beta. In this work, we demonstrate that the minimal genes required to produce aklanonic acid in heterologous hosts are dpsG (ACP), dauI (regulatory activator), dpsA (KS alpha), dpsB (KS beta), dpsF (aromatase), dpsE (polyketide reductase), and dauG (putative deoxyaklanonic acid oxygenase). The two unusual open reading frames, dpsC (KASIII homolog lacking a known active site) and dpsD (acyltransferase homolog), are not required to synthesize aklanonic acid. Additionally, replacement of dpsD or dpsCD in Streptomyces sp. strain C5 with a neomycin resistance gene (aphI) results in mutant strains that still produced anthracyclines. PMID:9098068

  16. An extremely alkaline mannanase from Streptomyces sp. CS428 hydrolyzes galactomannan producing series of mannooligosaccharides.

    PubMed

    Pradeep G C; Cho, Seung Sik; Choi, Yun Hee; Choi, Yun Seok; Jee, Jun-Pil; Seong, Chi Nam; Yoo, Jin Cheol

    2016-05-01

    An alkaline-thermostable mannanase from Streptomyces sp. CS428 was produced, purified, and biochemically characterized. The extracellular mannanase (Mn428) was purified to homogeneity with 12.4 fold, specific activity of 2406.7 U/mg, and final recovery of 37.6 %. The purified β-mannanase was found to be a monomeric protein with a molecular mass of approximately 35 kDa as analyzed by SDS-PAGE and zymography. The first N-terminal amino acid sequences of mannanase enzyme were HIRNGNHQLPTG. The optimal temperature and pH for enzyme were 60 °C and 12.5, respectively. The mannanase activities were significantly affected by the presence of metal ions, modulators, and detergents. Km and Vmax values of Mn428 were 1.01 ± 3.4 mg/mL and 5029 ± 85 µmol/min mg, respectively when different concentrations (0.6-10 mg/mL) of locust bean gum galactomannan were used as substrate. The substrate specificity of enzyme showed its highest specificity towards galactomannan which was further hydrolyzed to produce mannose, mannobiose, mannotriose, and a series of mannooligosaccharides. Mannooligosaccharides can be further converted to ethanol production, thus the purified β-mannanase isolated from Streptomyces sp. CS428 was found to be attractive for biotechnological applications.

  17. Mangrove Streptomyces sp. BDUKAS10 as nanofactory for fabrication of bactericidal silver nanoparticles.

    PubMed

    Sivalingam, Periyasamy; Antony, Jacob Joe; Siva, Durairaj; Achiraman, Shanmugam; Anbarasu, Kumarasamy

    2012-10-01

    Biosynthesis has led to the development of various biomimetic approaches for the fabrication of nanoscale materials. The present study reveals a unique procedure for the biosynthesis of bactericidal silver nanoparticles (AgNPs) using a novel Streptomyces sp. BDUKAS10, an isolate of mangrove sediment. Aqueous silver nitrate (AgNO(3)) solution was treated with cell free supernatant (CFS) of the isolate to synthesize bactericidal silver nanoparticles. Initial characterization was performed by visual observation for color change to intense brown color. UV-visible spectrophotometry (UV-vis) for measuring surface plasmon resonance indicated a maximum absorption peak at 441 nm. Fourier Transform Infrared Spectroscopy (FTIR) analysis provides evidence for proteins as possible reducing, and capping agents. Energy dispersive X-ray (EDAX) spectroscopy analysis showed elemental silver as major signal. Transmission Electron Microscopy (TEM) study indicated spherical silver nanoparticles in the size range of 21-48 nm. Compared to the CFS, the biosynthesized AgNPs exemplified superior bactericidal efficacy towards the tested bacterial strains. Results from this study suggested that Streptomyces sp. BDUKAS10 can be advantageous for the synthesis of AgNPs by extracellular method in the view of sustainable and ecofriendly approach.

  18. Sesquiterpenes and an intermediate 1alpha, 6beta, 11-eudesmanetriol in the biosynthesis of geosmin from Streptomyces sp.

    PubMed

    Yang, Ya-Bin; Yang, Zhi; Yang, Xue-Qiong; Zhang, Yong; Zhao, Li-Xing; Xu, Li-Hua; Ding, Zhong-Tao

    2012-03-01

    One new sesquiterpene was isolated from the fermentation broth of Streptomyces sp. and the structure was elucidated by spectral analysis as caryolane-1, 6beta-diol (1). An intermediate 1alpha, 6beta, 11-eudesmanetriol (2) in the biosynthesis of geosmin was also found in this strain which proved sequence for the reactions, especially bicyclization preceding dealkylation.

  19. Complete genome sequence of the Streptomyces sp. strain CdTB01, a bacterium tolerant to cadmium.

    PubMed

    Zhou, Geng; Yang, Hui; Zhou, Hui; Wang, Chong; Fu, Fuhua; Yu, Ye; Lu, Xiangyang; Tian, Yun

    2016-07-10

    Streptomyces sp. Strain CdTB01, which is tolerant to high concentrations of heavy metals, particularly cadmium, was isolated from soil contaminated with heavy metals. Two contigs with total genome size of 10.19Mb were identified in the whole genome sequencing and assembly, and numerous homologous genes known to be involved in heavy metal resistance were found in the genome.

  20. Isolation and characterization of soil Streptomyces species as potential biological control agents against fungal plant pathogens.

    PubMed

    Evangelista-Martínez, Zahaed

    2014-05-01

    The use of antagonist microorganisms against fungal plant pathogens is an attractive and ecologically alternative to the use of chemical pesticides. Streptomyces are beneficial soil bacteria and potential candidates for biocontrol agents. This study reports the isolation, characterization and antagonist activity of soil streptomycetes from the Los Petenes Biosphere Reserve, a Natural protected area in Campeche, Mexico. The results showed morphological, physiological and biochemical characterization of six actinomycetes and their inhibitory activity against Curvularia sp., Aspergillus niger, Helminthosporium sp. and Fusarium sp. One isolate, identified as Streptomyces sp. CACIS-1.16CA showed the potential to inhibit additional pathogens as Alternaria sp., Phytophthora capsici, Colletotrichum sp. and Rhizoctonia sp. with percentages ranging from 47 to 90 %. This study identified a streptomycete strain with a broad antagonist activity that could be used for biocontrol of plant pathogenic fungi.

  1. Evidence of α-, β- and γ-HCH mixture aerobic degradation by the native actinobacteria Streptomyces sp. M7.

    PubMed

    Sineli, P E; Tortella, G; Dávila Costa, J S; Benimeli, C S; Cuozzo, S A

    2016-05-01

    The organochlorine insecticide γ-hexachlorocyclohexane (γ-HCH, lindane) and its non-insecticidal α- and β-isomers continue to pose serious environmental and health concerns, although their use has been restricted or completely banned for decades. In this study we report the first evidence of the growth ability of a Streptomyces strain in a mineral salt medium containing high doses of α- and β-HCH (16.6 mg l(-1)) as a carbon source. Degradation of HCH isomers by Streptomyces sp. M7 was investigated after 1, 4, and 7 days of incubation, determining chloride ion release, and residues in the supernatants by GC with µECD detection. The results show that both the α- and β-HCH isomers were effectively metabolized by Streptomyces sp. M7, with 80 and 78 % degradation respectively, after 7 days of incubation. Moreover, pentachlorocyclohexenes and tetrachlorocyclohexenes were detected as metabolites. In addition, the formation of possible persistent compounds such as chlorobenzenes and chlorophenols were studied by GC-MS, while no phenolic compounds were detected. In conclusion, we have demonstrated for the first time that Streptomyces sp. M7 can degrade α- and β-isomers individually or combined with γ-HCH and could be considered as a potential agent for bioremediation of environments contaminated by organochlorine isomers.

  2. Comprehensive analysis of the cellulolytic system reveals its potential for deconstruction of lignocellulosic biomass in a novel Streptomyces sp.

    PubMed

    Pinheiro, Guilherme L; de Azevedo-Martins, Allan C; Albano, Rodolpho M; de Souza, Wanderley; Frases, Susana

    2017-01-01

    The giant snail Achatina fulica is considered an invasive species in most territories in which it was introduced, due to its ability to process a large amount of lignocellulose as a consequence of the presence of a cellulolytic-associated microflora. Streptomyces are well known as crucial agents in the decomposition of complex polymers in soil environments and also as cellulolytic symbionts commonly associated with herbivore insects. Here, we employed a combination of genomic and biochemical tools for a detailed evaluation of the cellulolytic potential of Streptomyces sp. I1.2, an aerobic bacterium isolated from the intestinal lumen of A. fulica in a screening for cellulolytic bacteria. Genomic analysis revealed that the ratio and diversity of CAZy domains and GH families coded by Streptomyces sp. I1.2 are comparable to those present in other highly cellulolytic bacteria. After growth on crystalline cellulose or sugarcane bagasse as sole carbon sources, the functionality of several genes encoding endoglucanases, cellobiohydrolases, xylanases, CBMs, and one β-glucosidase were confirmed by the combination of enzymatic activity measurements, zymography, TLC, and cellulose-binding assays. The endoglucanases secreted by this isolate were stable at 50 °C and exhibited activity over a broad pH range between 4.0 and 8.0. The endoglucanases and cellobiohydrolases secreted by Streptomyces sp. I1.2 exhibited specific activities that were similar to the levels present in a commercial cellulase preparation from Trichoderma reesei, while I1.2 xylanase levels were even 350 % higher. The results presented here show that Streptomyces sp. I1.2 is promising for future biotechnological applications, since it is able to produce endoglucanases, cellobiohydrolases, and xylanases in appreciable amounts when grown on a low-cost residue such as sugarcane bagasse.

  3. Structure and activity of lobophorins from a turrid mollusk-associated Streptomyces sp.

    PubMed

    Lin, Zhenjian; Koch, Michael; Pond, Christopher D; Mabeza, Gaiselle; Seronay, Romell A; Concepcion, Gisela P; Barrows, Louis R; Olivera, Baldomero M; Schmidt, Eric W

    2014-01-01

    A novel lumun-lumun sampling methodology was used to obtain a large diversity of micromollusks, including the new species Lienardia totopotens. In turn, from L. totopotens we cultivated a Streptomyces sp. strain that contained new and known spirotetronate polyketides, lobophorins (1-5). The structures were elucidated using spectroscopy, and the compounds were evaluated for cytotoxicity to human cells and activity against Mycobacterium tuberculosis, Bacillus subtilis, Pseudomonas aeruginosa and Burkholderia cepacia. Compounds 2-5 showed varying degrees of activity against human cells, M. tuberculosis and B. subtilis in the low μM to mid nM range but were inactive against the other strains, while 1 lacking digitoxose was inactive. Very slight structural changes in 2-5 led to varying antibacterial:cytotoxicity ratios, providing a possible basis to synthesize more selective derivatives.

  4. Lobophorins with antimycobacterial activity from a turrid mollusk-associated Streptomyces sp

    PubMed Central

    Lin, Zhenjian; Koch, Michael; Pond, Christopher D.; Mabeza, Gaiselle; Seronay, Romell A.; Concepcion, Gisela P.; Barrows, Louis R.; Olivera, Baldomero M.; Schmidt, Eric W.

    2015-01-01

    A novel lumun-lumun sampling methodology was used to obtain a large diversity of micromollusks, including the new species Lienardia totopotens. In turn, from L. totopotens we cultivated a Streptomyces sp. strain that contained new and known spirotetronate polyketides, lobophorins (1–5). The structures were elucidated using spectroscopy, and the compounds were evaluated for cytotoxicity to human cells and activity against Mycobacterium tuberculosis. A structure-activity relationship was discerned, wherein the lack of digitoxose in 1 led to lack of both cytotoxic and antibacterial activity. For compounds 2–5 both activities were in the low μM to mid nM range. Although this likely precludes their direct application in tuberculosis therapy due to possible poor therapeutic index, very slight changes in structure led to widely varying antibacterial:cytotoxicity ratios, providing a possible basis to synthesize more selective derivatives. PMID:24220110

  5. Aranciamycins I and J, Antimycobacterial Anthracyclines from an Australian Marine-Derived Streptomyces sp.

    PubMed

    Khalil, Zeinab G; Raju, Ritesh; Piggott, Andrew M; Salim, Angela A; Blumenthal, Antje; Capon, Robert J

    2015-04-24

    Chemical analysis of an Australian marine-derived Streptomyces sp. (CMB-M0150) yielded two new anthracycline antibiotics, aranciamycins I (1) and J (2), as well as the previously reported aranciamycin A (3) and aranciamycin (4). The aranciamycins 1-4, identified by detailed spectroscopic analysis, were noncytotoxic when tested against selected Gram-negative bacteria and fungi (IC50 >30 μM) and exhibited moderate and selective cytotoxicity against Gram-positive bacteria (IC50 >1.1 μM) and a panel of human cancer cell lines (IC50 > 7.5 μM). Significantly, 1-4 were cytotoxic (IC50 0.7-1.7 μM) against the Mycobacterium tuberculosis surrogate M. bovis bacille Calmette-Guérin.

  6. Antifungal Substances from Streptomyces sp. A3265 Antagonistic to Plant Pathogenic Fungi

    PubMed Central

    Van Minh, Nguyen; Woo, E-Eum; Kim, Ji-Yul; Kim, Dae-Won; Hwang, Byung Soon; Lee, Yoon-Ju; Lee, In-Kyoung

    2015-01-01

    In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria. PMID:26539051

  7. Ionofore antibiotic polynactin produced by Streptomyces sp. 156A isolated from Lake Baikal.

    PubMed

    Shishlyannikova, Tatyana A; Kuzmin, Anton V; Fedorova, Galina A; Shishlyannikov, Sergey M; Lipko, Irina A; Sukhanova, Elena V; Belkova, Natalia L

    2017-03-01

    The potential antibacterial activity of secondary metabolites produced by Streptomyces sp. 156A isolated from Lake Baikal was investigated. The selective liquid-liquid extraction method was applied to obtain a mixture of nactins (polynactin) produced by the strain. The polynactin consisted of nonactin (3%), monactin (18%), dinactin (36%), trinactin (31%) and tetranactin (12%). The compounds were identified by MS/MS, (1)H and (13)C NMR methods. The loss of neutral 184 and 198 Da fragments from a sodiated molecular ion, [M + Na](+), of nactins was observed in the MS/MS spectrum. The polynactin was shown to possess the antibiotic activity against Gram-positive strains including opportunistic strains and strains isolated from various ecosystems of Lake Baikal.

  8. Modeling of competitive mutualistic relationships. Application to cellulose degradation by Streptomyces sp. strains.

    PubMed

    Thierie, Jacques; Penninckx, Michel J

    2007-12-01

    A "cascade" model depicts microbial degradation of a complex nutrient/substrate through a succession of intermediate compounds. Each stage is characterized by a particular species producing a typical degradation enzyme induced by its own degradation product. The final compound of the cascade consists of a single assimilable substrate used by all species. This results in a competition situation, whereas the contribution of all strains to the production of a complete set of efficient enzymes generates a mutualistic relationship. The model was shown to be appropriate to describe degradation of cellulose by a consortium of Streptomyces sp. strains. The simplicity and the model capacity for generalization are promising and could be used for various degradation processes both at laboratory and environmental scales.

  9. Optimization of Inulinase Production from Garlic by Streptomyces sp. in Solid State Fermentation Using Statistical Designs

    PubMed Central

    Dilipkumar, M.; Rajasimman, M.; Rajamohan, N.

    2011-01-01

    Plackett-Burman design was employed for screening 18 nutrient components for the production of inulinase using Garlic as substrate by Streptomyces sp. in solid-state fermentation (SSF). From the experiments, 4 nutrients, namely, NH4NO3, MnSO4·7H2O, Soya bean cake, and K2HPO4 were found to be most significant nutrient components. Hence, these 4 components are selected. The selected components were optimized using response surface methodology (RSM). The optimum conditions are NH4NO3—6.63 mg/gds, MnSO4·7H2O—26.16 mg/gds, Soya bean cake—60.6 mg/gds, and K2HPO4—5.24 mg/gds. Under these conditions, the production of inulinase was found to be 76 U/gds. PMID:21541216

  10. Lower homologues of ahpatinin, aspartic protease inhibitors, from a marine Streptomyces sp.

    PubMed

    Sun, Yi; Takada, Kentaro; Nogi, Yuichi; Okada, Shigeru; Matsunaga, Shigeki

    2014-07-25

    Two linear peptides, ahpatinin Ac (1) and ahpatinin Pr (2), were isolated together with the known ahpatinin (i)Bu, pepstatin Ac, pepstatin Pr, and pepsinostreptin from a Streptomyces sp. derived from a deep-sea sediment. The structure of ahpatinin Pr (2) was assigned by interpretation of NMR data and HPLC analysis of the hydrolysate after converting to the DNP-L-Val derivative. During the LCMS analysis of the acid hydrolysate, products arising from the retro-aldol cleavage of the statine and Ahppa units in 2 were observed and could facilitate the determination of the absolute configuration of the statine class of nonproteinogenic amino acids. Both ahpatinin Ac (1) and ahpatinin Pr (2) potently inhibited pepsin and moderately inhibited cathepsin B.

  11. A new prenylated indole derivative from endophytic actinobacteria Streptomyces sp. neau-D50.

    PubMed

    Zhang, Ji; Wang, Ji-Dong; Liu, Chong-Xi; Yuan, Jia-Hui; Wang, Xiang-Jing; Xiang, Wen-Sheng

    2014-01-01

    A new prenylated indole derivative 3-acetonylidene-7-prenylindolin-2-one (1) was isolated from the endophytic actinobacterium Streptomyces sp. neau-D50, together with four known hybrid isoprenoids, 7-isoprenylindole-3-carboxylic acid (2), 3-cyanomethyl-6-prenylindole (3), 6-isoprenylindole-3-carboxylic acid (4) and 7,4'-dihydroxy-5-methoxy-8-(γ,γ-dimethylallyl)-flavanone (5). The structures of these compounds were elucidated on the basis of extensive spectroscopic analysis and comparison with data from the literature. Compounds 1 and 2 demonstrated strong cytotoxic activities against human lung adenocarcinoma cell line A549 with an IC50 of 3.3 and 5.1 μg mL(- 1), respectively, which are comparable to that of the positive control doxorubicin (4.2 μg mL(- 1)). Furthermore, compounds 1-4 exhibited potent antifungal activity against phytopathogenic fungi Colletotrichum orbiculare, Phytophthora capsici, Corynespora cassiicola and Fusarium oxysporum.

  12. Artificial Intelligence versus Statistical Modeling and Optimization of Cholesterol Oxidase Production by using Streptomyces Sp.

    PubMed Central

    Niwas, Ram; Osama, Khwaja; Khan, Saif; Haque, Shafiul; Tripathi, C. K. M.; Mishra, B. N.

    2015-01-01

    Cholesterol oxidase (COD) is a bi-functional FAD-containing oxidoreductase which catalyzes the oxidation of cholesterol into 4-cholesten-3-one. The wider biological functions and clinical applications of COD have urged the screening, isolation and characterization of newer microbes from diverse habitats as a source of COD and optimization and over-production of COD for various uses. The practicability of statistical/ artificial intelligence techniques, such as response surface methodology (RSM), artificial neural network (ANN) and genetic algorithm (GA) have been tested to optimize the medium composition for the production of COD from novel strain Streptomyces sp. NCIM 5500. All experiments were performed according to the five factor central composite design (CCD) and the generated data was analysed using RSM and ANN. GA was employed to optimize the models generated by RSM and ANN. Based upon the predicted COD concentration, the model developed with ANN was found to be superior to the model developed with RSM. The RSM-GA approach predicted maximum of 6.283 U/mL COD production, whereas the ANN-GA approach predicted a maximum of 9.93 U/mL COD concentration. The optimum concentrations of the medium variables predicted through ANN-GA approach were: 1.431 g/50 mL soybean, 1.389 g/50 mL maltose, 0.029 g/50 mL MgSO4, 0.45 g/50 mL NaCl and 2.235 ml/50 mL glycerol. The experimental COD concentration was concurrent with the GA predicted yield and led to 9.75 U/mL COD production, which was nearly two times higher than the yield (4.2 U/mL) obtained with the un-optimized medium. This is the very first time we are reporting the statistical versus artificial intelligence based modeling and optimization of COD production by Streptomyces sp. NCIM 5500. PMID:26368924

  13. Gene cloning, recombinant expression, purification and characterization of l-methionine decarboxylase from Streptomyces sp. 590.

    PubMed

    Hayashi, Masaya; Okada, Akane; Yamamoto, Kumiko; Okugochi, Tomomi; Kusaka, Chika; Kudou, Daizou; Nemoto, Michiko; Inagaki, Junko; Hirose, Yuu; Okajima, Toshihide; Tamura, Takashi; Soda, Kenji; Inagaki, Kenji

    2016-12-21

    l-Methionine decarboxylase (MetDC) from Streptomyces sp. 590 depends on pyridoxal 5'-phosphate and catalyzes the non-oxidative decarboxylation of l-methionine to produce 3-methylthiopropylamine and carbon dioxide. MetDC gene (mdc) was determined to consist of 1,674 bp encoding 557 amino acids, and the amino acid sequence is similar to that of l-histidine decarboxylases and l-valine decarboxylases from Streptomyces sp. strains. The mdc gene was cloned and recombinant MetDC was heterologously expressed by Escherichia coli The purification of recombinant MetDC was carried out by DEAE-Toyopearl and Ni-NTA agarose column chromatography. The recombinant enzyme was homodimeric with a molecular mass of 61,000 Da and showed optimal activity between 45 to 55 °C and at pH 6.6, and the stability below 30 °C and between pH 4.6 to 7.0. l-Methionine and l-norleucine were good substrates for MetDC. The Michaelis constants for l-methionine and l-norleucine were 30 and 73 mM, respectively. The recombinant MetDC (0.50 U/ml) severely inhibited growth of human tumour cells A431 (epidermoid ovarian carcinoma cell line) and MDA-MB-231 (breast cancer cell line), however showed relatively low cytotoxicity for human normal cell NHDF-Neo (dermal fibroblast cell line from neonatal foreskin). This study revealed the properties of the gene and the protein sequence of MetDC for the first time.

  14. Artificial Intelligence versus Statistical Modeling and Optimization of Cholesterol Oxidase Production by using Streptomyces Sp.

    PubMed

    Pathak, Lakshmi; Singh, Vineeta; Niwas, Ram; Osama, Khwaja; Khan, Saif; Haque, Shafiul; Tripathi, C K M; Mishra, B N

    2015-01-01

    Cholesterol oxidase (COD) is a bi-functional FAD-containing oxidoreductase which catalyzes the oxidation of cholesterol into 4-cholesten-3-one. The wider biological functions and clinical applications of COD have urged the screening, isolation and characterization of newer microbes from diverse habitats as a source of COD and optimization and over-production of COD for various uses. The practicability of statistical/ artificial intelligence techniques, such as response surface methodology (RSM), artificial neural network (ANN) and genetic algorithm (GA) have been tested to optimize the medium composition for the production of COD from novel strain Streptomyces sp. NCIM 5500. All experiments were performed according to the five factor central composite design (CCD) and the generated data was analysed using RSM and ANN. GA was employed to optimize the models generated by RSM and ANN. Based upon the predicted COD concentration, the model developed with ANN was found to be superior to the model developed with RSM. The RSM-GA approach predicted maximum of 6.283 U/mL COD production, whereas the ANN-GA approach predicted a maximum of 9.93 U/mL COD concentration. The optimum concentrations of the medium variables predicted through ANN-GA approach were: 1.431 g/50 mL soybean, 1.389 g/50 mL maltose, 0.029 g/50 mL MgSO4, 0.45 g/50 mL NaCl and 2.235 ml/50 mL glycerol. The experimental COD concentration was concurrent with the GA predicted yield and led to 9.75 U/mL COD production, which was nearly two times higher than the yield (4.2 U/mL) obtained with the un-optimized medium. This is the very first time we are reporting the statistical versus artificial intelligence based modeling and optimization of COD production by Streptomyces sp. NCIM 5500.

  15. Glucose(xylose) isomerase production by Streptomyces sp. CH7 grown on agricultural residues.

    PubMed

    Chanitnun, Kankiya; Pinphanichakarn, Pairoh

    2012-07-01

    Streptomyces sp. CH7 was found to efficiently produce glucose(xylose) isomerase when grown on either xylan or agricultural residues. This strain produced a glucose(xylose) isomerase activity of roughly 1.8 U/mg of protein when it was grown in medium containing 1% xylose as a carbon source. Maximal enzymatic activities of about 5 and 3 U/mg were obtained when 1% xylan and 2.5% corn husks were used, respectively. The enzyme was purified from a mycelial extract to 16-fold purity with only two consecutive column chromatography steps using Macro-prep DEAE and Sephacryl-300, respectively. The approximate molecular weight of the purified enzyme is 170 kDa, and it has four identical subunits of 43.6 kDa as estimated by SDS-PAGE. Its K m values for glucose and xylose were found to be 258.96 and 82.77 mM, respectively, and its V max values are 32.42 and 63.64 μM/min/mg, respectively. The purified enzyme is optimally active at 85°C and pH 7.0. It is stable at pH 5.5-8.5 and at temperatures up to 60°C after 30 min. These findings indicate that glucose(xylose) isomerase from Streptomyces sp. CH7 has the potential for industrial applications, especially for high-fructose syrup production and bioethanol fermentation from hemicellulosic hydrolysates by Saccharomyces cerevisiae.

  16. Nonomuraea monospora sp. nov., an actinomycete isolated from cave soil in Thailand, and emended description of the genus Nonomuraea.

    PubMed

    Nakaew, Nareeluk; Sungthong, Rungroch; Yokota, Akira; Lumyong, Saisamorn

    2012-12-01

    A novel actinomycete, designated strain PT708(T), was isolated from cave soil collected in Pha Tup Cave Forest Park, Nan province, Thailand. It produced compounds with antimicrobial and anticancer activities. Its chemotaxonomic properties were consistent with those of members of the genus Nonomuraea. The major menaquinone was MK-9(H(4)), with minor amounts of MK-9(H(6)), MK-9(H(2)), MK-10(H(2)) and MK-8(H(4)). The polar lipid profile contained phosphatidylmonomethylethanolamine, diphosphatidylglycerol, hydroxy-phosphatidylmonomethylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol mannoside and phosphatidylinositol. The major fatty acids were iso-C(16:0), 10-methyl C(17:0), C(16:0) and C(17:1)ω6c. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain PT708(T) belonged to the genus Nonomuraea and was most closely related to Nonomuraea rhizophila YIM 67092(T) (98.50% sequence similarity) and Nonomuraea rosea GW 12687(T) (98.30%). The genomic DNA G+C content of strain PT708(T) was 73.3 mol%. Unlike the recognized members of the genus Nonomuraea, the novel strain formed single spores at the tips of aerial hyphae. Based on the phenotypic, phylogenetic and genotypic evidence, strain PT708(T) represents a novel species of the genus Nonomuraea, for which the name Nonomuraea monospora sp. nov. is proposed. The type strain is PT708(T) ( = TISTR 1910(T) = JCM 16114(T)).

  17. Verrucosispora maris sp. nov., a novel deep-sea actinomycete isolated from a marine sediment which produces abyssomicins.

    PubMed

    Goodfellow, Michael; Stach, James E M; Brown, Roselyn; Bonda, Avinash Naga Venkata; Jones, Amanda L; Mexson, Joanne; Fiedler, Hans-Peter; Zucchi, Tiago Domingues; Bull, Alan T

    2012-01-01

    Verrucosispora isolate AB-18-032(T), the abyssomicin- and proximicin-producing actinomycete, has chemotaxonomic and morphological properties consistent with its classification in the genus Verrucosispora. The organism formed a distinct phyletic line in the Verrucosispora 16S rRNA gene tree sharing similarities of 99.7%, 98.7% and 98.9% with Verrucosispora gifhornensis DSM 44337(T), Verrucosispora lutea YIM 013(T) and Verrucosispora sediminis MS 426(T), respectively. It was readily distinguished from the two latter species using a range of phenotypic features and from V. gifhornensis DSM 44337(T), its nearest phylogenetic neighbor, by a DNA G+C content of 65.5 mol% obtained by thermal denaturation and fluorometry and DNA:DNA relatedness values of 64.0% and 65.0% using renaturation and fluorometric methods, respectively. It is apparent from the combined genotypic and phenotypic data that strain AB-18-032(T) should be classified in the genus Verrucosispora as a new species. The name Verrucosispora maris sp. nov. is proposed for this taxon with isolate AB-18-032(T) (= DSM 45365(T) = NRRL B-24793(T)) as the type strain.

  18. Streptomyces leeuwenhoekii sp. nov., the producer of chaxalactins and chaxamycins, forms a distinct branch in Streptomyces gene trees

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A polyphasic study was carried out to establish the taxonomic status of an Atacama Desert isolate, Streptomyces strain C34T, which synthesises novel antibiotics, the chaxalactins and chaxamycins. The organism was shown to have chemotaxonomic, cultural, and morphological properties consistent with it...

  19. Geodermatophilus poikilotrophi sp. nov.: A Multitolerant Actinomycete Isolated from Dolomitic Marble

    PubMed Central

    Montero-Calasanz, Maria del Carmen; Hofner, Benjamin; Göker, Markus; Rohde, Manfred; Spröer, Cathrin; Hezbri, Karima; Gtari, Maher; Schumann, Peter; Klenk, Hans-Peter

    2014-01-01

    A novel Gram-reaction-positive, aerobic actinobacterium, tolerant to mitomycin C, heavy metals, metalloids, hydrogen peroxide, desiccation, and ionizing- and UV-radiation, designated G18T, was isolated from dolomitic marble collected from outcrops in Samara (Namibia). The growth range was 15–35°C, at pH 5.5–9.5 and in presence of 1% NaCl, forming greenish-black coloured colonies on GYM Streptomyces agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for other representatives of the genus Geodermatophilus. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, and small amount of diphosphatidylglycerol. MK-9(H4) was the dominant menaquinone and galactose was detected as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids iso-C16:0 and iso-C15:0 and the unsaturated C17:1ω8c and C16:1ω7c. The 16S rRNA gene showed 97.4–99.1% sequence identity with the other representatives of genus Geodermatophilus. Based on phenotypic results and 16S rRNA gene sequence analysis, strain G18T is proposed to represent a novel species, Geodermatophilus poikilotrophi. Type strain is G18T (= DSM 44209T = CCUG 63018T). The INSDC accession number is HF970583. The novel R software package lethal was used to compute the lethal doses with confidence intervals resulting from tolerance experiments. PMID:25114928

  20. Heterologous expression of metK1-sp and afsR-sp in Streptomyces venezuelae for the production of pikromycin.

    PubMed

    Maharjan, Sushila; Oh, Tae-Jin; Lee, Hei Chan; Sohng, Jae Kyung

    2008-09-01

    Two regulator genes, metK1-sp and afsR-sp, from Streptomyces peucetius ATCC 27952 were heterologously expressed in S. venezuelae ATCC 15439, to produce 14-membered pikromycin antibiotics. The production of pikromycin was increased by 1.6-fold and 2.6-fold by the expression of metK1-sp and afsR-sp, respectively. The overexpression of metK1-sp and afsR-sp in S. venezuelae stimulated the expression of the pathway-specific regulatory genes, pikD and ketosynthase, as demonstrated by RT-PCR. The elevated transcripts of the pikD and ketosynthase genes were consistent with the enhanced production of pikromycin.

  1. Genome Sequence of Streptomyces caatingaensis CMAA 1322, a New Abiotic Stress-Tolerant Actinomycete Isolated from Dried Lake Bed Sediment in the Brazilian Caatinga Biome.

    PubMed

    Santos, Suikinai Nobre; Gacesa, Ranko; Taketani, Rodrigo Gouvêa; Long, Paul F; Melo, Itamar Soares

    2015-09-10

    The genome sequence of the first Streptomyces species isolated from the Brazilian Caatinga is reported here. Genes related to environmental stress tolerance were prevalent and included many secondary metabolic gene clusters.

  2. A promising strain of Streptomyces sp. with agricultural traits for growth promotion and disease management.

    PubMed

    Alam, Mansoor; Dharni, Seema; Abdul-Khaliq; Srivastava, Santosh Kumar; Samad, Abdul; Gupta, Mahesh Kumar

    2012-08-01

    A bacterial strain, Streptomyces sp. CIMAP- A1 was isolated from Geranium rhizosphere and identified by morphological, physiological, biochemical and molecular characters (16S rDNA gene sequence). Phylogenetically, it was found most closely related to S. vinacendrappus, strain NRRL-2363 with 99% sequence similarity. The strain had potential antagonistic activity (in vitro) against wide range of phytopathogenic fungi like Stemphylium sp., Botrytis cinerea, Sclerotinia sclerotiorum, Colletotrichum spp., Curvularia spp., Corynespora cassicola and Thielavia basicola. The extracellular secondary metabolites produced by the strain in the culture filtrates significantly inhibited the spore germination, growth of germ tube of the germinated spores and radial growth of Alternaria alternata, Colletotrichum acutatum, Curvularia andropogonis and Fusarium moniliforme. The extraction of culture filtrate with solvents and purification by following VLC and PTLC methods always yielded a 10th fraction antifungal compound showing activity against wide range of phytopathogenic fungi. The strain was able to produce siderophores and indole-3-acetic acid. The strain was found to enhance the growth and biomass production of Geranium. It increased 11.3% fresh shoot biomass of Geranium and 21.7% essential oil yield.

  3. Purification and biological evaluation of the metabolites produced by Streptomyces sp. TK-VL_333.

    PubMed

    Kavitha, Alapati; Prabhakar, Peddikotla; Vijayalakshmi, Muvva; Venkateswarlu, Yenamandra

    2010-06-01

    An Actinobacterium strain isolated from laterite soils of the Guntur region was identified as Streptomyces sp. TK-VL_333 by 16S rRNA analysis. Cultural, morphological and physiological characteristics of the strain were recorded. The secondary metabolites produced by the strain cultured on galactose-tyrosine broth were extracted and concentrated followed by defatting of the crude extract with cyclohexane to afford polar and non-polar residues. Purification of the two residues by column chromatography led to isolation of five polar and one non-polar fraction. Bioactivity- guided fractions were rechromatographed on a silica gel column to obtain four compounds, namely 1H-indole-3-carboxylic acid, 2,3-dihydroxy-5-(hydroxymethyl) benzaldehyde, 4-(4-hydroxyphenoxy) butan-2-one and acetic acid-2-hydroxy-6-(3-oxo-butyl)-phenyl ester from three active polar fractions and 8-methyl decanoic acid from one non-polar fraction. The structure of the compounds was elucidated on the basis of FT-IR, mass and NMR spectroscopy. The antimicrobial activity of the bioactive compounds produced by the strain was tested against the bacteria and fungi and expressed in terms of minimum inhibitory concentration. Antifungal activity of indole-3-carboxylic acid was further evaluated under in vitro and in vivo conditions. This is the first report of 2,3-dihydroxy-5-(hydroxymethyl) benzaldehyde, 4-(4-hydroxyphenoxy) butan-2-one, acetic acid-2-hydroxy-6-(3-oxo-butyl)-phenyl ester and 8-methyl decanoic acid from the genus Streptomyces.

  4. Antifungal and antibacterial activities of Streptomyces polymachus sp. nov. isolated from soil.

    PubMed

    Nguyen, Tuan Manh; Kim, Jaisoo

    2015-08-01

    Strain T258T was isolated from forest soil at Bongnae Falls, South Korea. The strain exhibited antimicrobial and antifungal activity against the following strains: Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Paenibacillus larvae, Escherichia coli, Candida albicans and Aspergillus niger. Growth occurred on all ISP media tested (2, 3, 4, 5, 6 and 7), Czapek-Dox agar, potato dextrose agar, trypticase soy agar, Bennett's modified agar and nutrient agar at 28 °C. Aerial spores were produced solely on ISP Medium 4; the colour of the aerial mycelium was white and the substrate mycelium was ivory. Melanin production was negative on peptone-yeast extract iron agar (ISP Medium 6). The cell-wall peptidoglycan contained ll-diaminopimelic acid, glutamic acid, alanine and glycine. Whole-cell hydrolysates contained glucose, ribose and galactose. The predominant menaquinones were MK-9(H6) and MK-9(H8) while the minor menaquinone was MK-10(H2). The polar lipids included diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The major fatty acids (>10%) were C16 : 0 (29.8%), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (15.1%), anteiso-C15 : 0 (13.5%) and iso-C15 : 0 (10.3%). DNA-DNA similarity with other strains ranged between 37.84 ± 1.15% and 50.25 ± 1.91 %. On the basis of these data, we suggest that strain T258T represents a novel species that belong to the genus Streptomyces, for which we propose a name Streptomyces polymachus sp. nov. The type strain is T258T ( = KACC 18247T = KEMB 9005-212T = NBRC 110905T).

  5. Sulfotanone, a new alkyl sulfonic acid derivative from Streptomyces sp. IFM 11694 with TRAIL resistance-overcoming activity.

    PubMed

    Abdelfattah, Mohamed S; Ishikawa, Naoki; Karmakar, Utpal K; Ishibashi, Masami

    2016-04-01

    One new alkyl sulfonic acid derivative, sulfotanone (1), and the known panosialin wA (2) were isolated from the methanolic extract of mycelium of Streptomyces sp. 11694. The structure of the new compound (1) was established by a combination of spectroscopic techniques, including HRESIMS, IR, 1D and 2D NMR measurements. Compound 1 (40 µM) in combination with TRAIL showed synergistic activity in sensitizing TRAIL-resistance in human gastric adenocarcinoma cell lines.

  6. Maniwamycins: new quorum-sensing inhibitors against Chromobacterium violaceum CV026 were isolated from Streptomyces sp. TOHO-M025.

    PubMed

    Fukumoto, Atsushi; Murakami, Chikana; Anzai, Yojiro; Kato, Fumio

    2016-05-01

    Quorum sensing is an important microbial signaling system that controls the expression of many virulence genes. Maniwamycins C-F, new compounds and quorum-sensing inhibitors, were isolated from the culture broth of Streptomyces sp. TOHO-M025 using a silica gel column and preparative HPLC. The structures of maniwamycins were elucidated by spectroscopic analyses, including NMR. The compounds each have an azoxy moiety. All maniwamycins inhibited violacein synthesis, which is controlled by quorum sensing, in Chromobacterium violaceum CV026.

  7. Extracellular biosynthesis of silver nanoparticle using Streptomyces sp. 09 PBT 005 and its antibacterial and cytotoxic properties

    NASA Astrophysics Data System (ADS)

    Saravana Kumar, P.; Balachandran, C.; Duraipandiyan, V.; Ramasamy, D.; Ignacimuthu, S.; Al-Dhabi, Naif Abdullah

    2015-02-01

    The application of microorganisms for the synthesis of nanoparticles as an eco-friendly and promising approach is welcome due to its non-toxicity and simplicity. The aim of this study was to synthesize silver nanoparticle using Streptomyces sp. (09 PBT 005). 09 PBT 005 was isolated from the soil sample of the agriculture field in Vengodu, Thiruvannamalai district, Tamil Nadu, India. 09 PBT 005 was subjected to molecular characterization by 16S rRNA sequence analysis. It was found that 09 PBT 005 belonged to Streptomyces sp. The isolate Streptomyces sp. 09 PBT 005 was inoculated in fermentation medium and incubated at 30 ºC for 12 days in different pH conditions. The 0.02 molar concentration showed good antibacterial activity against Gram-positive and Gram-negative bacteria at pH-7. The synthesis of silver nanoparticles was investigated by UV-Vis spectroscopy, scanning electron microscopy and Fourier Transform Infrared analysis. The synthesized AgNPs sizes were found to be in the dimensions ranging between 198 and 595 nm. The cytotoxicity of the synthesized nanoparticles was studied against A549 adenocarcinoma lung cancer cell line. It showed 83.23 % activity at 100 μl with IC 50 value of 50 μl. This method will be useful in the biosynthesis of nanoparticles.

  8. Identification and characterization of a Streptomyces sp. isolate exhibiting activity against multidrug-resistant coagulase-negative Staphylococci.

    PubMed

    Sadigh-Eteghad, Saeed; Dehnad, Alireza; Shanebandi, Dariush; Khalili, Iraj; Razmarayii, Nasser; Namvaran, Ali

    2011-12-01

    The resistance of 220 coagulase-negative Staphylococci (CNS) (associated with animal disease) to 13 antibiotics were determined using the disk diffusion method. 35.9% of multidrug-resistant coagulase-negative Staphylococci (MR-CNS) exhibited resistance to five or more than five antibiotics; all of these bacteria were resistant to methicillin too. The new Streptomyces sp. ABRIINW111 was isolated from the Zagros Mountains Hamadan, Iran. The 16S rDNA sequence of the isolate indicated that it has 98% similarity to S. levis, but some mutations in the alpha and gamma regions of the 16S rDNA sequence emphasize the probability of the existence of a new species. Preliminary and secondary antibacterial screenings revealed that the isolate is active against gram negative and positive bacteria. The diethyl ether extracted metabolite of the Streptomyces sp. ABRIINW111 showed an effective antibacterial activity against MR-CNS. So the diethyl ether extract of the new Streptomyces sp. strain ABRIINW111 can inhibit the MR-CNS in vitro, and it can offer a new approach to treat MR-CNS infectious patients.

  9. Micromonospora taraxaci sp. nov., a novel endophytic actinomycete isolated from dandelion root (Taraxacum mongolicum Hand.-Mazz.).

    PubMed

    Zhao, Junwei; Guo, Lifeng; He, Hairong; Liu, Chongxi; Zhang, Yuejing; Li, Chuang; Wang, Xiangjing; Xiang, Wensheng

    2014-10-01

    A novel actinomycete, designated strain NEAU-P5(T), was isolated from dandelion root (Taraxacum mongolicum Hand.-Mazz.). Strain NEAU-P5(T) showed closest 16S rRNA gene sequence similarity to Micromonospora chokoriensis 2-19/6(T) (99.5%), and phylogenetically clustered with Micromonospora violae NEAU-zh8(T) (99.3%), M. saelicesensis Lupac 09(T) (99.0%), M. lupini Lupac 14N(T) (98.8%), M. zeae NEAU-gq9(T) (98.4%), M. jinlongensis NEAU-GRX11(T) (98.3%) and M. zamorensis CR38(T) (97.9%). Phylogenetic analysis based on the gyrB gene sequence also indicated that the isolate clustered with the above type strains except M. violae NEAU-zh8(T). The cell-wall peptidoglycan consisted of meso-diaminopimelic acid and glycine. The major menaquinones were MK-9(H8), MK-9(H6) and MK-10(H2). The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The major fatty acids were C(16:0), iso-C(15:0) and C(17:0). Furthermore, some physiological and biochemical properties and low DNA-DNA relatedness values enabled the strain to be differentiated from members of closely related species. Therefore, it is proposed that strain NEAU-P5(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora taraxaci sp. nov. is proposed. The type strain is NEAU-P5(T) (=CGMCC 4.7098(T) = DSM 45885(T)).

  10. Actinomycetospora rhizophila sp. nov., an actinomycete isolated from rhizosphere soil of a peace lily (Spathi phyllum Kochii).

    PubMed

    He, Hairong; Zhang, Yuejing; Ma, Zhaoxu; Li, Chuang; Liu, Chongxi; Zhou, Ying; Li, Lianjie; Wang, Xiangjing; Xiang, Wensheng

    2015-05-01

    A novel actinomycete, designated strain NEAU-B-8(T), was isolated from the rhizosphere soil of a peace lily (Spathi phyllum Kochii) collected from Heilongjiang province, north-east China. Key morphological and physiological characteristics as well as chemotaxonomic features of strain NEAU-B-8(T) were congruent with the description of the genus Actinomycetospora , such as the major fatty acids, the whole-cell hydrolysates, the predominant menaquinone and the phospholipid profile. The 16S rRNA gene sequence analysis revealed that strain NEAU-B-8(T) shared the highest sequence similarities with Actinomycetospora lutea JCM 17982(T) (99.3% 16S rRNA gene sequence similarity), Actinomycetospora chlora TT07I-57(T) (98.4 %), Actinomycetospora straminea IY07-55(T) (98.3%) and Actinomycetospora chibensis TT04-21(T) (98.2%); similarities to type strains of other species of this genus were lower than 98%. The phylogenetic tree based on 16S rRNA gene sequences showed that strain NEAU-B-8(T) formed a distinct branch with A. lutea JCM 17982(T) that was supported by a high bootstrap value of 97% in the neighbour-joining tree and was also recovered with the maximum-likelihood algorithm. However, the DNA-DNA relatedness between strain NEAU-B-8(T) and A. lutea JCM 17982(T) was found to be 50.6 ± 1.2%. Meanwhile, strain NEAU-B-8(T) differs from other most closely related strains in phenotypic properties, such as maximum NaCl tolerance, hydrolysis of aesculin and decomposition of urea. On the basis of the morphological, physiological, chemotaxonomic, phylogenetic and DNA-DNA hybridization data, we conclude that strain NEAU-B-8(T) represents a novel species of the genus Actinomycetospora , named Actinomycetospora rhizophila sp. nov. The type strain is NEAU-B-8(T). ( = CGMCC 4.7134(T) =DSM 46673(T)).

  11. Cryptoendolithic actinomycetes from antarctic sandstone rock samples: Micromonospora endolithica sp. nov. and two isolates related to Micromonospora coerulea Jensen 1932.

    PubMed

    Hirsch, Peter; Mevs, Udo; Kroppenstedt, Reiner M; Schumann, Peter; Stackebrandt, Erko

    2004-03-01

    Three cryptoendolithic, aerobic actinomycetes (AA-459T, AA-319 and AA-321) from antarctic sandstone were characterised phenotypically and by molecular taxonomic methods. The isolates had single spores on substrate mycelium, meso-diaminopimelic acid (m-DAP) and glycine (cell wall type II), a whole cell sugar pattern D (galactose, xylose, arabinose, glucose or rhamnose) and phospholipids of type PII (diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol). Their predominant fatty acids were iso-16:0 and iso-15:0 or 17:1omega8c, the menaquinone profile was complex with mainly MK10 (H4) and MK10 (H6). A wide variety of sugars and several acids were utilised for growth. The isolates were sensitive to a few antibiotics, but formation and excretion of antibiotics was not observed. Phenotypically, isolates AA-319 and AA-321 were similar. Phylogenetic analysis of 16S rRNA gene sequences revealed close relationship of strains AA-319 and AA-321 with each other (99.5%) and clustering (98.5%) with Micromonospora coerulea DSM 43143T. DNA-DNA hybridisation showed both strains to be genomically highly similar to strain DSM 43143T. Phenotypically they could be viewed as separate taxa, but presently they will be considered as strains of Micromonospora coerulea. Strain AA-459T was phylogenetically close to Micromonospora chersina DSM 44151T (99.1%) and to Micromonospora rosaria DSM 803T, but DNA-DNA similarity with M. chersina DSM 44151T was low with 28.9/33.5 %, indicating the presence of a different and new species. Consequently, isolate AA-459T (DSM 44398T NRRL B-24248T) is described as the type strain of Micromonospora endolithica sp. nov.

  12. Actinomadura syzygii sp. nov., an endophytic actinomycete isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels).

    PubMed

    Rachniyom, Hathairat; Matsumoto, Atsuko; Indananda, Chantra; Duangmal, Kannika; Takahashi, Yoko; Thamchaipenet, Arinthip

    2015-06-01

    The taxonomic position of an endophytic actinomycete, strain GKU 157T, isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels) collected at Khao Khitchakut National Park, Chantaburi province, Thailand, was determined using a polyphasic taxonomic approach. 16S rRNA gene sequence analysis revealed that strain GKU 157T belongs to the genus Actinomadura and formed a distinct phyletic line with Actinomadura chibensis NBRC 106107T (98.6 % similarity). Strain GKU 157T formed an extensively branched, non-fragmenting substrate mycelium and aerial hyphae that differentiated into hooked to short spiral chains of about 20 non-motile spores with a warty surface. The cell wall contained meso-diaminopimelic acid and the whole-cell sugars were galactose, glucose, madurose, mannose and ribose. The N-acyl type of muramic acid was acetyl. Mycolic acids were absent. The phospholipids included phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylinositol (PI), phosphatidylinositolmannoside (PIM) and two unknown phospholipids (PLs). The major menaquinone was MK-9(H6) and the predominant fatty acids were C16:0, iso-C16:0, C18:1ω9c, C18:0 and 10-methyl C18:0 (tuberculostearic acid). The genomic DNA G+C content was 73.1 mol%. A combination of DNA-DNA hybridization results and significant differences from related species in cultural, physiological and chemical characteristics indicated that strain GKU 157T represents a novel species of the genus Actinomadura, for which the name Actinomadura syzygii sp. nov. is proposed. The type strain is GKU 157T ( = BCC 70456T = NBRC 110399T).

  13. Nonomuraea syzygii sp. nov., an endophytic actinomycete isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels).

    PubMed

    Rachniyom, Hathairat; Matsumoto, Atsuko; Indananda, Chantra; Duangmal, Kannika; Takahashi, Yoko; Thamchaipenet, Arinthip

    2015-04-01

    A novel endophytic actinomycete, designated strain GKU 164(T), was isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels), collected at Khao Khitchakut National Park, Chantaburi province, Thailand. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain formed a distinct clade within the genus Nonomuraea , and was most closely related to Nonomuraea monospora PT708(T) (98.77% 16S rRNA gene sequence similarity) and Nonomuraea thailandensis KC-061(T) (98.73%). Strain GKU 164(T) formed a branched substrate and aerial hyphae that generated single spores with rough surfaces. The cell wall contained meso-diaminopimelic acid. The whole-cell sugars were madurose, galactose, mannose, ribose, rhamnose and glucose. The N-acyl type of muramic acid was acetyl. The predominant menaquinone was MK-9(H4) with minor amounts of MK-9(H6), MK-9(H2) and MK-9(H0). The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositolmannosides, phosphatidylmonomethylethanolamine, hydroxy-phosphatidylmonomethylethanolamine, an unidentified aminophosphoglycolipid and four unknown phospholipids. The major fatty acids were iso-C(16 : 0) and 10-methyl C(17 : 0). The genomic DNA G+C content was 70.4 mol%. Significant differences in the morphological, chemotaxonomical, and biochemical data together with DNA-DNA relatedness values between strain GKU 164(T) and type strains of closely related species, clearly demonstrated that strain GKU 164(T) represents a novel species of the genus Nonomuraea , for which the name Nonomuraea syzygii sp. nov. is proposed. The type strain is GKU 164(T) ( = BCC 70457(T) = NBRC 110400(T)).

  14. Improvement of FK506 Production in the High-Yielding Strain Streptomyces sp. RM7011 by Engineering the Supply of Allylmalonyl-CoA Through a Combination of Genetic and Chemical Approach.

    PubMed

    Mo, SangJoon; Lee, Sung-Kwon; Jin, Ying-Yu; Suh, Joo-Won

    2016-02-01

    FK506, a widely used immunosuppressant, is a 23-membered polyketide macrolide that is produced by several Streptomyces species. FK506 high-yielding strain Streptomyces sp. RM7011 was developed from the discovered Streptomyces sp. KCCM 11116P by random mutagenesis in our previous study. The results of transcript expression analysis showed that the transcription levels of tcsA, B, C, and D were increased in Streptomyces sp. RM7011 by 2.1-, 3.1-, 3.3-, and 4.1- fold, respectively, compared with Streptomyces sp. KCCM 11116P. The overexpression of tcsABCD genes in Streptomyces sp. RM7011 gave rise to approximately 2.5-fold (238.1 μg/ml) increase in the level of FK506 production compared with that of Streptomyces sp. RM7011. When vinyl pentanoate was added into the culture broth of Streptomyces sp. RM7011, the level of FK506 production was approximately 2.2-fold (207.7 μg/ml) higher than that of the unsupplemented fermentation. Furthermore, supplementing the culture broth of Streptomyces sp. RM7011 expressing tcsABCD genes with vinyl pentanoate resulted in an additional 1.7-fold improvement in the FK506 titer (498.1 μg/ml) compared with that observed under nonsupplemented condition. Overall, the level of FK506 production was increased approximately 5.2-fold by engineering the supply of allylmalonyl-CoA in the high-yielding strain Streptomyces sp. RM7011, using a combination of overexpressing tcsABCD genes and adding vinyl pentanoate, as compared with Streptomyces sp. RM7011 (95.3 μg/ml). Moreover, among the three precursors analyzed, pentanoate was the most effective precursor, supporting the highest titer of FK506 in the FK506 high-yielding strain Streptomyces sp. RM7011.

  15. Aerobic and microaerophilic actinomycetes of typical agropeat and peat soils

    NASA Astrophysics Data System (ADS)

    Zenova, G. M.; Gryadunova, A. A.; Pozdnyakov, A. I.; Zvyagintsev, D. G.

    2008-02-01

    A high number (from tens of thousands to millions of CFU/g of soil) of actinomycetes and a high diversity of genera were found in typical peat and agropeat soils. Agricultural use increases the number and diversity of the actinomycete complexes of the peat soils. In the peat soils, the actinomycete complex is represented by eight genera: Streptomyces, Micromonospora, Streptosporangium, Actinomadura, Microbispora, Saccharopolyspora, Saccharomonospora, and Microtetraspora. A considerable share of sporangial forms in the actinomycete complex of the peat soils not characteristic of the zonal soils was revealed. The number of actinomycetes that develop under aerobic conditions is smaller by 10-100 times than that of aerobic forms in the peat soils. Among the soil actinomycetes of the genera Streptomyces, Micromonospora, Streptosporangium, Actinomadura, Microbispora, and Microtetraspora, the microaerophilic forms were found; among the Saccharopolyspora and Saccharomonospora, no microaerophilic representatives were revealed.

  16. Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov., two novel actinomycetes isolated from muddy soil and emended description of the genus Actinomadura.

    PubMed

    Zhao, Junwei; Guo, Lifeng; Sun, Pengyu; Han, Chuanyu; Bai, Lu; Liu, Chongxi; Li, Yunxi; Xiang, Wensheng; Wang, Xiangjing

    2015-12-01

    Two novel actinomycetes, designated strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T), were isolated from muddy soil collected from a riverbank in Jiaohe, Jilin Province, north China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibited 99.9 % 16S rRNA gene sequence similarity with each other and that they are closely related to Actinomadura viridis IFO 15238(T) (99.6, 99.6 %) and Actinomadura vinacea IFO 14688(T) (99.3, 99.3 %). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the two cultures clustered together and formed a cluster with A. viridis IFO 15238(T), A. vinacea IFO 14688(T) and Actinomadura rugatobispora IFO 14382(T). However, the DNA-DNA hybridization value between strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) was 63.6 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) represent two novel species of the genus Actinomadura, for which the names Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov. are proposed. The type strains are NEAU-Jh1-3(T) (=CGMCC 4.7197(T) = JCM 30341(T)) and NEAU-Jh2-5(T) (=CGMCC 4.7211(T) = JCM 30342(T)), respectively.

  17. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

    PubMed Central

    Fleige, Christian; Meyer, Florian

    2016-01-01

    ABSTRACT The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation resulted in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement was achieved with constitutive expression of the vanillin anabolism genes ech and fcs, coding for the enzymes feruloyl-coenzyme A (CoA) synthetase (fcs) and enoyl-CoA hydratase/aldolase (ech). The transcription of both genes was shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin was efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase was eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, was obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter was achieved, at the expense of a lower molar yield, by using an improved feeding strategy. This is the highest reported vanillin concentration reached in microbial fermentation processes without extraction of the product. Furthermore, the vanillin was produced almost without by-products, with a molar yield that nearly approached the theoretical maximum. IMPORTANCE Much effort has been put into optimization of the biotechnological production of natural vanillin. The demand for this compound is growing due to increased consumer concerns regarding chemically produced food additives. Since this

  18. Amycolatopsis rhabdoformis sp. nov., an actinomycete isolated from a tropical forest soil.

    PubMed

    Souza, Wallace Rafael; Silva, Rafael Eduardo; Goodfellow, Michael; Busarakam, Kanungnid; Figueiro, Fernanda Sales; Ferreira, Douglas; Rodrigues-Filho, Edson; Moraes, Luiz Alberto Beraldo; Zucchi, Tiago Domingues

    2015-06-01

    Strain SB026T was isolated from Brazilian rainforest soil and its taxonomic position established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological features consistent with its classification in the genus Amycolatopsis and formed a branch in the Amycolatopsis 16S rRNA gene tree together with Amycolatopsis bullii NRRL B-24847T, Amycolatopsis plumensis NRRL B-24324T, Amycolatopsis tolypomycina DSM 44544T and Amycolatopsis vancoresmycina NRRL B-24208T. It was related most closely to A. bullii NRRL B-24847T (99.0 % 16S rRNA gene sequence similarity), but was distinguished from this strain by a low level of DNA-DNA relatedness (~46 %) and discriminatory phenotypic properties. Based on the combined genotypic and phenotypic data, it is proposed that the isolate should be classified in the genus Amycolatopsis as representing a novel species, Amycolatopsis rhabdoformis sp. nov. The type strain is SB026T ( = CBMAI 1694T = CMAA 1285T = NCIMB 14900T).

  19. Purification and characterisation of an acidic and antifungal chitinase produced by a Streptomyces sp.

    PubMed

    Karthik, Narayanan; Binod, Parameswaran; Pandey, Ashok

    2015-01-01

    An extremely acidic extracellular chitinase produced by a Streptomyces sp. was purified 12.44-fold by ammonium sulphate precipitation, ion-exchange chromatography and gel-permeation chromatography and further characterised. The molecular mass of the enzyme was estimated to be about 40 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 2 and 6, and 50 °C respectively. The enzyme showed high stability in the acidic pH range of 2-6 and temperature stability of up to 50 °C. Additionally, the effect of some cations and other chemical compounds on the chitinase activity was studied. The activity of the enzyme was considerably retained under salinity conditions of up to 3%. The Km and Vmax values of the enzyme were determined to be 6.74 mg mL(-1) and 61.3 U mg(-1) respectively using colloidal chitin. This enzyme exhibited antifungal activity against phytopathogens revealing a potential biocontrol application in agriculture.

  20. Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007.

    PubMed

    Yuan, Dongjuan; Lan, Dongming; Xin, Ruipu; Yang, Bo; Wang, Yonghua

    2016-01-01

    A screening method along with the combination of genome sequence of microorganism, pairwise alignment, and lipase classification was used to search the thermostable lipase. Then, a potential thermostable lipase (named MAS1) from marine Streptomyces sp. strain W007 was expressed in Pichia pastoris X-33, and the biochemical properties were characterized. Lipase MAS1 belongs to the subfamily I.7, and it has 38% identity to the well-characterized Bacillus subtilis thermostable lipases in the subfamily I.4. The purified enzyme was estimated to be 29 kDa. The enzyme showed optimal temperature at 40 °C, and retained more than 80% of initial activity after 1 H incubation at 60 °C, suggesting that MAS1 was a thermostable lipase. MAS1 was an alkaline enzyme with optimal pH value at 7.0 and had stable activity for 12 H of incubation at pH 6.0-9.0. It was stable and retained about 90% of initial activity in the presence of Cu(2+) , Ca(2+) , Ni(2+) , and Mg(2+) , whereas 89.05% of the initial activity was retained when ethylene diamine tetraacetic acid was added. MAS1 showed the tolerance to organic solvents, but was inhibited by various surfactants. MAS1 was verified to be a triglyceride lipase and could hydrolyze triacylglycerol and diacylglycerol. The result represents a good example for researchers to discover thermostable lipase for industrial application.

  1. Production of polysaccharide-based bioflocculant for the synthesis of silver nanoparticles by Streptomyces sp.

    PubMed

    Manivasagan, Panchanathan; Kang, Kyong-Hwa; Kim, Dong Gyu; Kim, Se-Kwon

    2015-01-01

    Polysaccharide-based bioflocculants have attracted considerable attention in recent years due to their biodegradable, harmless and negligible secondary pollution. Bioflocculants are organic macromolecular substances secreted by microorganisms. A simple, cost-effective and green method was developed for the biosynthesis of silver nanoparticles using polysaccharides as reducing and stabilizing agents. In this paper, we report on the production and optimization of polysaccharide-based bioflocculant for the green synthesis of silver nanoparticles by Streptomyces sp. MBRC-91. Medium composition and culture conditions for polysaccharide-based bioflocculants were statistically optimized by response surface methodology (RSM). The bioflocculant production was statistically optimized with most significant factors, namely palm jaggery (18.73g/L), yeast extract (2.07g/L), K2HPO4 (3.74g/L) and NaCl (0.38g/L), respectively. The biosynthesized silver nanoparticles were characterized by UV-vis spectroscopy, XRD, FTIR, FESEM, EDXA and HRTEM. The biosynthesized silver nanoparticles revealed strong antibacterial activity in sewage water and this result could make a new avenue in the wastewater treatment. Therefore, the biosynthesized silver nanoparticles can be extended as an alternative for the development of new bactericidal bionanomaterials for wastewater treatment and biotechnological applications.

  2. HAF, hepatoma aggregation factor produced by Streptomyces sp. strain No. A-6143.

    PubMed

    Suzuki, K; Nakano, N; Nagatomi, Y; Tominaga, H; Nakazono, N; Itai, M; Uyeda, M; Shibata, M

    1990-08-01

    We searched for a new cell aggregation factor for hepatoma AH109A cells, and found one we called HAF in the culture filtrate of Streptomyces sp. strain No. A-6143 isolated from a soil sample. HAF was purified by salting-out with ammonium sulfate. DEAE-cellulose column chromatography, gel filtration on Sephadex G-100, and hydroxylapatite column chromatography, HAF was glycoprotein which had a molecular weight of about 73,000. HAF was stable from pH 6 to 8 at 37 degrees C and up to 40 degrees C at pH 8.0 and the aggregation activity of HAF was maximum around pH 8 at 30 degrees C. The activity was not influenced by some saccharides, but it was inhibited by EDTA and EGTA: moreover HAF activity was restored by the addition of calcium ions. HAF aggregated hepatoma AH136B and COS-7 cells as well as hepatoma AH109A cells, but it was inert to other cancer cells and human erythrocytes. These properties proved that HAF is completely different from other aggregation factors for cancer cells so far reported.

  3. Identification of Elaiophylin Skeletal Variants from the Indonesian Streptomyces sp. ICBB 9297.

    PubMed

    Sheng, Yan; Lam, Phillip W; Shahab, Salmah; Santosa, Dwi Andreas; Proteau, Philip J; Zabriskie, T Mark; Mahmud, Taifo

    2015-11-25

    Four new elaiophylin macrolides (1-4), together with five known elaiophylins (5-9), have been isolated from cultures of the Indonesian soil bacterium Streptomyces sp. ICBB 9297. The new compounds have macrocyclic skeletons distinct from those of the known dimeric elaiophylins in that one or both of the polyketide chains contain(s) an additional pendant methyl group. Further investigations revealed that 1 and 2 were derived from 3 and 4, respectively, during isolation processes. Compounds 1-3 showed comparable antibacterial activity to elaiophylin against Staphylococcus aureus. However, interestingly, only compounds 1 and 3, which contain a pendant methyl group at C-2, showed activity against Mycobacterium smegmatis, whereas compound 2, which has two pendant methyl groups at C-2 and C-2', and the known elaiophylin analogues (5-7), which lack pendant methyl groups at C-2 and/or C-2', showed no activity. The production of 3 and 4 in strain ICBB 9297 indicates that one of the acyltransferase (AT) domains in the elaiophylin polyketide synthases (PKSs) can recruit both malonyl-CoA and methylmalonyl-CoA as substrates. Bioinformatic analysis of the AT domains of the elaiophylin PKSs revealed that the ela_AT7 domain contains atypical active site amino acid residues, distinct from those conserved in malonyl-CoA- or methylmalonyl-CoA-specific ATs.

  4. Stereochemistry and conformation of skyllamycin, a non-ribosomally synthesized peptide from Streptomyces sp. Acta 2897.

    PubMed

    Schubert, Vivien; Di Meo, Florent; Saaidi, Pierre-Loïc; Bartoschek, Stefan; Fiedler, Hans-Peter; Trouillas, Patrick; Süssmuth, Roderich D

    2014-04-22

    Skyllamycin is a non-ribosomally synthesized cyclic depsipeptide from Streptomyces sp. Acta 2897 that inhibits PDGF-signaling. The peptide scaffold contains an N-terminal cinnamoyl moiety, a β-methylation of aspartic acid, three β-hydroxylated amino acids and one rarely occurring α-hydroxy glycine. With the exception of α-hydroxy glycine, the stereochemistry of the amino acids was assigned by comparison to synthetic reference amino acids applying chiral GC-MS and Marfey-HPLC analysis. The stereochemistry of α-hydroxy glycine, which is unstable under basic and acidic conditions, was determined by conformational analysis, employing a combination of data from NOESY-NMR spectroscopy, simulated annealing and free MD simulations. The simulation procedures were applied for both R- and S-configured α-hydroxy glycine of the skyllamycin structure and compared to the NOESY data. Both methods, simulated annealing and free MD simulations independently support S-configured α-hydroxy glycine thus enabling the assignment of all stereocenters in the structure of skyllamycin and devising the role of two-component flavin dependent monooxygenase (Sky39) as S-selective.

  5. Purification and crystallographic analysis of a FAD-dependent halogenase from Streptomyces sp. JCM9888.

    PubMed

    Zhao, Yanqun; Yan, Baohua; Yang, Ting; Jiang, Jian; Wei, Heng; Zhu, Xiaofeng

    2015-08-01

    A new FAD (flavin adenine dinucleotide)-dependent halogenase HalY from Streptomyces sp. JCM9888 was reported to be involved in the regioselective halogenation of adenine. HalY is a variant B FAD-dependent halogenase that is most similar to the halogenase PltA involved in pyoluteorin biosynthesis. This study reports the overexpression and purification of HalY with an N-terminal hexahistidine tag, followed by crystallization experiments and X-ray crystallographic analysis. HalY was purified as a monomer in solution and crystallized to give X-ray diffraction to a resolution of 1.7 Å. The crystal belonged to the monoclinic space group P21, with unit-cell parameters a = 41.4, b = 113.4, c = 47.6 Å, α = γ = 90, β = 107.4°, and contained one monomer of HalY in the asymmetric unit, with a calculated Matthews coefficient of 2.3 Å(3) Da(-1) and a solvent content of 46%. The structure of the halogenase CndH was used as a search model in molecular replacement to obtain the initial model of HalY. Manual model building and structure refinement of HalY are in progress.

  6. Streptomyces sp. LK3 mediated synthesis of silver nanoparticles and its biomedical application.

    PubMed

    Karthik, L; Kumar, Gaurav; Kirthi, A Vishnu; Rahuman, A A; Bhaskara Rao, K V

    2014-02-01

    In the present study, the marine actinobacteria mediated biosynthesis of silver nanoparticles (AgNps) was achieved using Streptomyces sp LK3. The synthesized AgNps showed the characteristic absorption spectra in UV-vis at 420 nm, which confirmed the presence of nanoparticles. XRD analysis showed intense peaks at 2θ values of 27.51°, 31.87°, 45.57°, 56.56°, 66.26°, and 75.25° corresponding to (210), (113), (124), (240), (226), and (300) Bragg's reflection based on the fcc structure of AgNps. The FTIR spectra exhibited prominent peaks at 3,417 cm(-1) (OH stretching due to alcoholic group) and 1,578 cm(-1) (C=C ring stretching). TEM micrograph showed that the synthesized AgNps were spherical in shape with an average size of 5 nm. Surface morphology and topographical structure of the synthesized AgNps were dignified by AFM. The synthesized AgNps showed significant acaricidal activity against Rhipicephalus microplus and Haemaphysalis bispinosa with LC50 values of 16.10 and 16.45 mg/L, respectively. Our results clearly indicate that AgNps could provide a safer alternative to conventional acaricidal agents in the form of a topical antiparasitic formulation. The present study aimed to develop a novel, cost-effective, eco-friendly actinobacteria mediated synthesis of AgNps and its antiparasitic activity.

  7. Production of α-amylase for the biosynthesis of gold nanoparticles using Streptomyces sp. MBRC-82.

    PubMed

    Manivasagan, Panchanathan; Venkatesan, Jayachandran; Kang, Kyong-Hwa; Sivakumar, Kannan; Park, Sun-Joo; Kim, Se-Kwon

    2015-01-01

    Marine actinobacterial synthesis of gold nanoparticles has good potential to develop simple, cost-effective and eco-friendly methods for production of important biomaterials. In this context, gold nanoparticles have attracted considerable attention in recent years, owing to their various applications. In this paper, we report on the production of α-amylase for the extracellular synthesis of gold nanoparticles using Streptomyces sp. MBRC-82. Medium composition and culture conditions for α-amylase production were statistically optimized. Plackett-Burman design was employed to find out the optimal medium constituents and culture conditions to enhance α-amylase production. Box-Behnken design revealed that three independent variables namely soluble starch (5.8484 g), peptone (3.5191 g), and NaCl (0.3829) significantly influenced α-amylase production. The gold nanoparticles were characterized by ultraviolet-visible (UV-vis) spectrometer, X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDXA), and transmission electron microscopy (TEM). The particles synthesized using the optimized enzyme activity ranged from 20 to 80 nm with an average particle size of 40 nm and therefore can be extended to various medicinal applications.

  8. Rhodococcus canchipurensis sp. nov., an actinomycete isolated from a limestone deposit site.

    PubMed

    Nimaichand, Salam; Sanasam, Suchitra; Zheng, Liu-Qiang; Zhu, Wen-Yong; Yang, Ling-Ling; Tang, Shu-Kun; Ningthoujam, Debananda S; Li, Wen-Jun

    2013-01-01

    A novel actinobacterial strain, MBRL 353(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. Comparison of 16S rRNA gene sequences of strain MBRL 353(T) and other members of the genus Rhodococcus showed sequence similarities ranging from 95.5 to 98.2 %, with strain MBRL 353(T) showing closest sequence similarity to Rhodococcus triatomae IMMIB RIV-085(T) (98.2 %) and Rhodococcus equi DSM 20307(T) (97.2 %). DNA-DNA hybridization results, however, revealed that DNA-DNA relatedness values between strain MBRL 353(T) and R. triatomae DSM 44892(T) (43.4 %) and R. equi DSM 20307(T) (33.4 %) were well below the 70 % limit for species identification. Strain MBRL 353(T) contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose in the cell wall. Mycolic acids were present. The major fatty acids were C(16 : 0) (45.7 %), C(18 : 1)ω9c (18.2 %) and 10-methyl C(18 : 0) (11.3 %). The only menaquinone detected was MK-8(H(2)), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 69.2 mol%. The phenotypic and genotypic data showed that strain MBRL 353(T) merits recognition as a representative of a novel species of the genus Rhodococcus for which the name Rhodococcus canchipurensis sp. nov. is proposed; the type strain is MBRL 353(T) (= KCTC 19851(T) = JCM 17578(T)).

  9. Pseudonocardia nantongensis sp. nov., a novel endophytic actinomycete isolated from the coastal halophyte Tamarix chinensis Lour.

    PubMed

    Xing, Ke; Qin, Sheng; Bian, Guang-Kai; Zhang, Yue-Ji; Zhang, Wen-Di; Dai, Chuan-Chao; Liu, Chang-Hong; Li, Wen-Jun; Jiang, Ji-Hong

    2012-11-01

    A novel isolate, designated strain KLBMP 1282(T) was isolated from the surface-sterilized leaves of a coastal halophyte Tamarix chinensis Lour., collected from Nantong, Jiangsu Province, east of China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that this strain belongs to the genus Pseudonocardia, being most closely related to Pseudonocardia kongjuensis LM 157(T) (98.33 %), Pseudonocardia autotrophica IMSNU 20050(T) (97.77 %), Pseudonocardia endophytica YIM 56035(T) (97.63 %), Pseudonocardia ammonioxydans H9 (T) (97.62 %) and Pseudonocardia compacta IMSNU 20111(T) (97.56 %); similarity to other type strains of the genus Pseudonocardia was <97.5 %. Chemotaxonomic data confirmed the affiliation of strain KLBMP 1282(T) to the genus Pseudonocardia. Strain KLBMP 1282(T) contained MK-8(H(4)) as the predominant ubiquinone and iso-C(16:0) as the major fatty acid. The polar lipids detected in strain KLBMP 1282(T) were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, one unknown phospholipid and four unknown glycolipids. The DNA G + C content of strain KLBMP 1282(T) was 73.1 mol %. The results of DNA-DNA hybridizations and the phylogenetic analysis, together with the phenotypic and biochemical tests, allowed the differentiation of strain KLBMP 1282(T) from strains of other recognized Pseudonocardia species. Therefore, strain KLBMP 1282(T) represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia nantongensis sp. nov. is proposed. The type strain is KLBMP 1282(T) (=KCTC 29053(T) = NBRC 108677(T)).

  10. Cloning and sequencing of a gene encoding a novel extracellular neutral proteinase from Streptomyces sp. strain C5 and expression of the gene in Streptomyces lividans 1326.

    PubMed Central

    Lampel, J S; Aphale, J S; Lampel, K A; Strohl, W R

    1992-01-01

    The gene encoding a novel milk protein-hydrolyzing proteinase was cloned on a 6.56-kb SstI fragment from Streptomyces sp. strain C5 genomic DNA into Streptomyces lividans 1326 by using the plasmid vector pIJ702. The gene encoding the small neutral proteinase (snpA) was located within a 2.6-kb BamHI-SstI restriction fragment that was partially sequenced. The molecular mass of the deduced amino acid sequence of the mature protein was determined to be 15,740, which corresponds very closely with the relative molecular mass of the purified protein (15,500) determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified neutral proteinase was determined, and the DNA encoding this sequence was found to be located within the sequenced DNA. The deduced amino acid sequence contains a conserved zinc binding site, although secondary ligand binding and active sites typical of thermolysinlike metalloproteinases are absent. The combination of its small size, deduced amino acid sequence, and substrate and inhibition profile indicate that snpA encodes a novel neutral proteinase. Images PMID:1569011

  11. Halophilic and halotolerant actinomycetes from a marine saltern of Goa, India producing anti-bacterial metabolites.

    PubMed

    Ballav, Shuvankar; Kerkar, Savita; Thomas, Sabu; Augustine, Nimmy

    2015-03-01

    Marine salterns are estuarine ecosystems in Goa, receiving inputs from riverine and marine waters. The Salinity fluctuates between 0 and 300 psu which makes it a conducive niche for salt tolerant and salt loving Actinomycetales. Halotolerant and halophilic Actinomycetales producing anti-bacterial metabolites were studied from crystallizer pond sediments of Ribandar saltern, Goa. Three media viz. Starch casein, R2A and Inorganic salt starch agar at four different salinities (35, 50, 75 and 100 psu) were used for isolation. R2A agar at 35 psu was the most preferred by hypersaline actinomycetes. The dominant group was halotolerant Streptomyces spp. others being rare actinomycetes viz. Nocardiopsis, Micromonospora and Kocuria spp. More than 50% of the isolates showed anti-bacterial activity against one or more of the fifteen human pathogens tested. Eight strains from 4 genera showed consistent anti-bacterial activity and studied in detail. Most halotolerant isolates grew from 0 to 75 psu, with optimum antibiotic production at 35 psu whereas halophiles grew at 20 to 100 psu with optimum antibiotic production at 35 psu. Four Streptomyces strains showed multiple inhibition against test organisms while four rare actinomycetes were specific in their inhibitory activity. This is the first report of a halophilic Kocuria sp., Nocardiopsis sp., and halotolerant Micromonospora sp. producing anti-bacterial compound(s) against Staphylococcus aureus, Staphylococcus citreus, and Vibrio cholerae, respectively. Sequential extraction with varying polarity of organic solvents showed that the extracts inhibited different test pathogens. These results suggest that halophilic and halotolerant actinomycetes from marine salterns are a potential source of anti-bacterial compounds.

  12. Lobophorins E and F, new spirotetronate antibiotics from a South China Sea-derived Streptomyces sp. SCSIO 01127.

    PubMed

    Niu, Siwen; Li, Sumei; Chen, Yuchan; Tian, Xinpeng; Zhang, Haibo; Zhang, Guangtao; Zhang, Weimin; Yang, Xiaohong; Zhang, Si; Ju, Jianhua; Zhang, Changsheng

    2011-11-01

    The strain SCSIO 01127, isolated from the South China Sea sediment, was identified as a member of Streptomyces by the 16S rDNA sequence analysis. Two new spirotetronate antibiotics lobophorins E (1) and F (2), along with two known analogs lobophorins A (3) and B (4), were isolated from Streptomyces sp. SCSIO 01127. Their structures were elucidated on the basis of detailed IR, NMR and MS spectroscopic analyses. The new compound lobophorin F (2) showed antibacterial activities against Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 with MIC values of 8 μg ml(-1) for both the strains, better than that of lobophorin B (4). Lobophorin F (2) also displayed better cytotoxic activities than lobophorin B (4), with IC(50) of 6.82, 2.93 and 3.16 μM against SF-268, MCF-7 and NCI-H460, respectively.

  13. Streptomyces yangpuensis sp. nov., a novel streptomycete isolated from soil in Shanghai, China.

    PubMed

    Tang, Biao; Yu, Yucong; Zhi, Xiaoyang; Yang, Lingling; Cen, Xufeng; Zhao, Guoping; Ding, Xiaoming

    2015-12-23

    A novel Gram-positive strain with sandy aerial mycelium and golden yellow substrate mycelium, designated fd2-tbT, was isolated from a soil sample collected in Shanghai, China, and its taxonomic status was established by phylogenetic analysis. Analysis showed that the nearly complete 16S rRNA gene sequence of fd2-tbT belonged to the genus Streptomyces, and was related to S. amritsarensis JCM19660T (99.9 %), S. flavotricini NBRC 12770T (99.9 %), S. polychromogenes NBRC 13072T (99.8 %), S. racemochromogenes NRRL B-5430T (99.7 %), S. globosus LMG 19896T (99.5 %), S. toxytricini NBRC 12823T (99.5 %) and S. katrae NBRC 13447T (99.3 %). The fd2-tbT cell wall contained LL-diaminopimelic acid, and whole cell sugars were identified as glucose and ribose. The menaquinones MK-9(H4), MK-9(H6) and MK-9(H8) were also detected. In addition, the polar lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, as well as five unidentified phospholipids, were detected. Major cellular fatty acids were identified as anteiso-C15:0, iso-C16:0, anteiso-C17:0, iso-C15:0 and C16:0. DNA-DNA hybridization experiments showed that strain fd2-tbT exhibited 36.5±0.6 %, 43.5±2.0 %, 11.1±1.3 %, 10.3±3.1 %, 9.8±1.9 %, 48.9±3.9 % and 16.3±1.7 % relatedness to S. amritsarensis JCM19660T, S. flavotricini NBRC 12770T, S. polychromogenes NBRC 13072T, S. racemochromogenes NRRL B-5430T, S. globosus LMG 19896T, S. toxytricini NBRC 12823T and S. katrae NBRC 13447T, respectively. Based on these analyses as well as some phenotypic differences, strain fd2-tbT is considered to represent a novel species, for which the name Streptomyces yangpuensis sp. nov. is proposed. The type strain is fd2-tbT (=DSM 100336T = CGMCC 4.7256T).

  14. Endophytic actinomycetes from spontaneous plants of Algerian Sahara: indole-3-acetic acid production and tomato plants growth promoting activity.

    PubMed

    Goudjal, Yacine; Toumatia, Omrane; Sabaou, Nasserdine; Barakate, Mustapha; Mathieu, Florence; Zitouni, Abdelghani

    2013-10-01

    Twenty-seven endophytic actinomycete strains were isolated from five spontaneous plants well adapted to the poor sandy soil and arid climatic conditions of the Algerian Sahara. Morphological and chemotaxonomical analysis indicated that twenty-two isolates belonged to the Streptomyces genus and the remaining five were non-Streptomyces. All endophytic strains were screened for their ability to produce indole-3-acetic acid (IAA) in vitro on a chemically defined medium. Eighteen strains were able to produce IAA and the maximum production occurred with the Streptomyces sp. PT2 strain. The IAA produced was further extracted, partially purified and confirmed by thin layer chromatography (TLC) analysis. The 16S rDNA sequence analysis and phylogenetic studies indicated that strain PT2 was closely related to Streptomyces enissocaecilis NRRL B 16365(T), Streptomyces rochei NBRC 12908(T) and Streptomyces plicatus NBRC 13071(T), with 99.52 % similarity. The production of IAA was affected by cultural conditions such as temperature, pH, incubation period and L-tryptophan concentration. The highest level of IAA production (127 μg/ml) was obtained by cultivating the Streptomyces sp. PT2 strain in yeast extract-tryptone broth supplemented with 5 mg L-tryptophan/ml at pH 7 and incubated on a rotary shaker (200 rpm) at 30 °C for 5 days. Twenty-four-hour treatment of tomato cv. Marmande seeds with the supernatant culture of Streptomyces sp. PT2 that contained the crude IAA showed the maximum effect in promoting seed germination and root elongation.

  15. Actinomycetes in the rhizosphere of semidesert soils of Mongolia

    NASA Astrophysics Data System (ADS)

    Norovsuren, Zh.; Zenova, G. M.; Mosina, L. V.

    2007-04-01

    The population density of actinomycetes in the desert-steppe soil, rhizosphere, and the above-ground parts of plants varies from tens to hundreds of thousands of colony-forming units (CFU) per gram of substrate. The actinomycetal complexes of the brown desert-steppe soil without plant roots are more diverse in their taxonomic composition than the actinomycetal complexes in the rhizosphere and the aboveground parts of plants. Additionally to representatives of the Streptomyces and Micromonospora genera, actinomycetes from the Nocardia, Saccharopolyspora, Thermomonospora, and Actinomadura genera were identified in the soil. The population density of actinomycetes in the rhizosphere and in the soil reached hundreds of thousand CFU/g; it considerably exceeded the population density of actinomycetes in the aboveground parts of plants. The maximum population density of actinomycetes was determined in the rhizosphere of Asparagus gobicus, Salsola pestifera, and Cleistogenes songorica.

  16. Diversity and biosynthetic potential of culturable actinomycetes associated with marine sponges in the China Seas.

    PubMed

    Xi, Lijun; Ruan, Jisheng; Huang, Ying

    2012-01-01

    The diversity and secondary metabolite potential of culturable actinomycetes associated with eight different marine sponges collected from the South China Sea and the Yellow sea were investigated. A total of 327 strains were isolated and 108 representative isolates were selected for phylogenetic analysis. Ten families and 13 genera of Actinomycetales were detected, among which five genera represent first records isolated from marine sponges. Oligotrophic medium M5 (water agar) proved to be efficient for selective isolation, and "Micromonospora-Streptomyces" was proposed as the major distribution group of sponge-associated actinomycetes from the China Seas. Ten isolates are likely to represent novel species. Sponge Hymeniacidon perleve was found to contain the highest genus diversity (seven genera) of actinomycetes. Housekeeping gene phylogenetic analyses of the isolates indicated one ubiquitous Micromonospora species, one unique Streptomyces species and one unique Verrucosispora phylogroup. Of the isolates, 27.5% displayed antimicrobial activity, and 91% contained polyketide synthase and/or nonribosomal peptide synthetase genes, indicating that these isolates had a high potential to produce secondary metabolites. The isolates from sponge Axinella sp. contained the highest presence of both antimicrobial activity and NRPS genes, while those from isolation medium DNBA showed the highest presence of antimicrobial activity and PKS I genes.

  17. Inhibition of norsolorinic acid accumulation to Aspergillus parasiticus by marine actinomycetes

    NASA Astrophysics Data System (ADS)

    Yan, Peisheng; Shi, Cuijuan; Shen, Jihong; Wang, Kai; Gao, Xiujun; Li, Ping

    2014-11-01

    Thirty-six strains of marine actinomycetes were isolated from a sample of marine sediment collected from the Yellow Sea and evaluated in terms of their inhibitory activity on the growth of Aspergillus parasiticus and the production of norsolorinic acid using dual culture plate assay and agar diffusion methods. Among them, three strains showed strong antifungal activity and were subsequently identified as Streptomyces sp. by 16S rRNA gene sequencing analysis. The supernatant from the fermentation of the MA01 strain was extracted sequentially with chloroform and ethyl acetate, and the activities of the extracts were determined by tip culture assay. The assay results show that both extracts inhibited mycelium growth and toxin production, and the inhibitory activities of the extracts increased as their concentrations increased. The results of this study suggest that marine actinomycetes are biologically important for the control of mycotoxins, and that these bacteria could be used as novel biopesticides against mycotoxins.

  18. Purification, Characterization, and Substrate Specificities of Multiple Xylanases from Streptomyces sp. Strain B-12-2

    PubMed Central

    Elegir, Graziano; Szakács, George; Jeffries, Thomas W.

    1994-01-01

    The endoxylanase complex from Streptomyces sp. strain B-12-2 was purified and characterized. The organism forms five distinct xylanases in the absence of significant cellulase activity when grown on oat spelt xylan. This is the largest number of endoxylanases yet reported for a streptomycete. On the basis of their physiochemical characteristics, they can be divided into two groups: the first group (xyl 1a and xyl 1b) consists of low-molecular-mass (26.4 and 23.8 kDa, respectively) neutral- to high-pI (6.5 and 8.3, respectively) endoxylanases. Group 1 endoxylanases are unable to hydrolyze aryl-β-d-cellobioside, have low levels of activity against xylotetraose (X4) and limited activity against xylopentaose, produce little or no xylose, and form products having a higher degree of polymerization with complex substrates. These enzymes apparently carry out transglycosylation. The second group (xyl 2, xyl 3, and xyl 4) consists of high-molecular-mass (36.2, 36.2, and 40.5 kDa, respectively), low-pI (5.4, 5.0, and 4.8, respectively) xylanases. Group 2 endoxylanases are able to hydrolyze aryl-β-d-cellobioside, show higher levels of activity against X4, and hydrolyze xylopentaose completely with the formation of xylobiose and xylotriose plus limited amounts of X4 and xylose. The enzymes display intergroup synergism when acting on kraft pulp. Despite intragroup similarities, each enzyme exhibited a unique action pattern and physiochemical characteristic. xyl 2 was highly glycosylated, and xyl 1b (but no other enzyme) was completely inhibited by p-hydroxymercuribenzoate. Images PMID:16349337

  19. Production, Characterization and Antioxidant Potential of Protease from Streptomyces sp. MAB18 Using Poultry Wastes

    PubMed Central

    Manivasagan, Panchanathan; Sivakumar, Kannan; Kim, Se-Kwon

    2013-01-01

    Poultry waste is an abundant renewable source for the recovery of several value-added metabolites with potential industrial applications. This study describes the production of protease on poultry waste, with the subsequent use of the same poultry waste for the extraction of antioxidants. An extracellular protease-producing strain was isolated from Cuddalore coast, India, and identified as Streptomyces sp. MAB18. Its protease was purified 17.13-fold with 21.62% yield with a specific activity of 2398.36 U/mg and the molecular weight was estimated as 43 kDa. The enzyme was optimally active at pH 8–10 and temperature 50–60°C and it was most stable up to pH 12 and 6–12% of NaCl concentration. The enzyme activity was reduced when treated with Hg2+, Pb2+, and SDS and stimulated by Fe2+, Mg2+, Triton X-100, DMSO (dimethyl sulfoxide), sodium sulphite, and β-mercaptoethanol. Furthermore, the antioxidant activities of protease were evaluated using in vitro antioxidant assays, such as DPPH radical-scavenging activity, O2 scavenging activity, NO scavenging activity, Fe2+ chelating activity, and reducing power. The enzyme showed important antioxidant potential with an IC50 value of 78 ± 0.28 mg/mL. Results of the present study indicate that the poultry waste-derived protease may be useful as supplementary protein and antioxidant in the animal feed formulations. PMID:23991418

  20. Determination of 2-methylisoborneol and geosmin produced by Streptomyces sp. and Anabaena PCC7120.

    PubMed

    Xie, Yuqun; He, Jin; Huang, Jun; Zhang, Jibin; Yu, Ziniu

    2007-08-22

    A new sample preparation and enrichment technique, headspace liquid-phase microextraction (HS-LPME) linked to gas chromatography-mass spectrometry (GC-MS), was developed for the determination of the off-flavor odorants, 2-methylisoborneol and geosmin, produced by Streptomyces sp. and Anabaena PCC7120. Some of the factors that influence the extraction efficiency of HS-LPME, such as the type of extraction solvent, ionic strength of sample solution, and sample agitation rate, were studied and optimized by a single factor test. Other factors, including extraction temperature, extraction time, microdrop volume, and headspace volume were optimized by orthogonal array design. Extraction of 2-methylisoborneol and geosmin was conducted by exposing 2.5 microL of 1-hexanol for 9 min at 50 degrees C in the headspace of a 20 mL vial with a 10 mL of sample solution saturated by NaCl and stirred at 800 rpm. The developed protocol demonstrated good repeatability (relative standard deviations (RSDs) < 5%), wide linear ranges (10-5000 ng/L, r2 > 0.999), and low limits of detection (LODs) for 2-methylisoborneol and geosmin (0.05 ng/L for both analytes). Subsequently, the method was successfully applied to extract the analytes in bacterial cultures with high recoveries (from 94% to 98%). Compared with headspace solid-phase microextraction (HS-SPME), HS-LPME demonstrates better linearity, precision, and recovery. Importantly, the sensitivity is about 1 order of magnitude higher than that of most HS-SPME. The results showed that HS-LPME coupled with GC-MS is a simple, convenient, rapid, sensitive, and effective method for the qualitative and quantitative analysis of 2-methylisoborneol and geosmin.

  1. Purification and Characterization of a Novel Extracellular Thermostable Alkaline Protease from Streptomyces sp. M30.

    PubMed

    Xin, Yan; Sun, Zhibin; Chen, Qiongzhen; Wang, Jue; Wang, Yicheng; Luogong, Linfeng; Li, Shuhuan; Dong, Weiliang; Cui, Zhongli; Huang, Yan

    2015-11-01

    A novel alkaline protease from Streptomyces sp. M30, SapHM, was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and DEAE-Sepharose chromatography, with a yield of 15.5% and a specific activity of 29,070 U/mg. Tryptic fragments of the purified SapHM were obtained by electrospray ionization quadrupole time-of- flight mass spectrometry. Nucleotide sequence analysis revealed that the gene sapHM contained 1,179 bp, corresponding to 392 amino acids with conserved Asp156, His187, and Ser339 residues of alkaline protease. The first 24 amino acid residues were predicted to be a signal peptide, and the molecular mass of the mature peptide was 37.1 kDa based on amino acid sequences and mass spectrometry. Pure SapHM was optimally active at 80°C in 50 mM glycine-NaOH buffer (pH 9.0), and was broadly stable at 0-50 °C and pH 4.0-9.0. The protease relative activity was increased in the presence of Ni(2+), Mn(2+), and Cu(2+) to 112%, 113%, and 147% of control, respectively. Pure SapHM was also activated by dimethylformamide, dimethyl sulfoxide, Tween 80, and urea. The activity of the purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. The Km and Vmax values were estimated to be 35.7 mg/ml, and 5 × 10(4) U/mg for casein. Substrate specificity analysis showed that SapH was active on casein, bovine serum albumin, and bovine serum fibrin.

  2. 12T061A and 12T061C, two new julichrome family compounds, as radical scavengers from Streptomyces sp.

    PubMed

    Komoda, Toshikazu; Saeki, Naoko; Koseki, Yoshitaka; Kiyota, Hiromasa

    2016-01-01

    We identified two new radical scavengers, 12T061A (1, C19H20O7) and 12T061C (2, C20H22O7), from a culture of the Streptomyces sp. Spectroscopic analysis indicated that these compounds are new julichrome family compounds. Compounds 1 and 2 showed radical-scavenging activity with an ED50 of 370 μM and 18 μM, respectively. Moreover, 1 showed tumor cell growth suppressive activity in HepG2 cells, (IC50: 3.6 μM); however, no suppressive activity was shown in 2 (IC50: > 100 μM).

  3. Anti-dormant mycobacterial activity and target analysis of nybomycin produced by a marine-derived Streptomyces sp.

    PubMed

    Arai, Masayoshi; Kamiya, Kentaro; Pruksakorn, Patamaporn; Sumii, Yuji; Kotoku, Naoyuki; Joubert, Jean-Pierre; Moodley, Prashini; Han, Chisu; Shin, Dayoung; Kobayashi, Motomasa

    2015-07-01

    In the course of our search for anti-dormant Mycobacterial substances, nybomycin (1) was re-discovered from the culture broth of a marine-derived Streptomyces sp. on the bioassay-guided separation. Compound 1 showed anti-microbial activity against Mycobacterium smegmatis and Mycobacterium bovis BCG with the MIC of 1.0μg/mL under both actively growing aerobic conditions and dormancy inducing hypoxic conditions. Compound 1 is also effective to Mycobacterium tuberculosis including the clinically isolated strains. The mechanistic analysis indicated that 1 bound to DNA and induces a unique morphological change to mycobacterial bacilli leading the bacterial cell death.

  4. Antifungal properties of an actinomycin D-producing strain, Streptomyces sp. IA1, isolated from a Saharan soil.

    PubMed

    Toumatia, Omrane; Yekkour, Amine; Goudjal, Yacine; Riba, Amar; Coppel, Yannick; Mathieu, Florence; Sabaou, Nasserdine; Zitouni, Abdelghani

    2015-02-01

    An actinomycete strain named IA1, which produced an antimicrobial compound, was isolated from a Saharan soil in In Amenas, Algeria. The study of the 16S rDNA sequence of this strain permitted to relate it to Streptomyces mutabilis NBRC 12800(T) (99.93% of similarity). Strain IA1 exhibited strong activity against a wide range of plant pathogenic fungi. One bioactive compound produced in large amounts (46.7 mg L(-1)  day(-1) ), named YA, was isolated and purified by TLC and reverse phase HPLC. The structure elucidation of the pure substance, using combined data from UV visible, NMR spectra, and mass spectrometry, permitted to identify it as actinomycin D, and was thus found for the first time in S. mutabilis related species. The biocontrol abilities of the strain IA1 and compound YA were evaluated through two diseases, i.e., chocolate spot of field bean and Fusarium wilt of flax. The occurrence of the two fungal diseases was effectively reduced. The reduction of chocolate spot disease symptoms reached 80 and 91.7% with IA1 and YA seedlings pretreatments, respectively. Soil pretreatment with IA1 or YA also allowed to reduce Fusarium wilt disease impact by almost 60%.

  5. Elimination of indigenous linear plasmids in Streptomyces hygroscopicus var. jinggangensis and Streptomyces sp. FR008 to increase validamycin A and candicidin productivities.

    PubMed

    Lu, Chenyang; Wu, Hang; Su, Xiurong; Bai, Linquan

    2017-02-25

    Giant linear plasmids, which replicate independently of the chromosomes, widely exist in actinobacteria. Previous studies mostly focused on the replication and evolution of the linear plasmids or the secondary metabolite gene clusters and the resistance gene clusters therein. However, the relationships of the linear plasmids to the productivities of secondary metabolites have not been studied. In this work, we developed a method to eliminate the indigenous linear plasmid pSHJG1 in Streptomyces hygroscopicus var. jinggangensis, and validamycin A titer increased by 12.5% (from 19.16 ± 1.93 to 21.56 ± 2.25 g/L) in the high-yielding strain TL01 and 43.7% (from 4.67 ± 0.05 to 6.71 ± 0.21 g/L) in the wild-type strain 5008, whereas the cellular growth of the plasmid-cured mutant was reduced. Subsequently, the plasmid-cured mutant was complemented with three structure genes involved in cellular growth in pSHJG1 under the control of a strong PvalA promoter. Among them, the complementation of genes pSHJG1.069 and pSHJG1.072, encoding a putative hydrolase and putative P-loop ATPase, respectively, resulted in the restoration of cellular growth and validamycin A titer. Furthermore, the elimination of indigenous linear plasmid pHZ228 in the candicidin producer Streptomyces sp. FR008 also led to enhanced candicidin production and reduced cellular growth. Because of the wide distribution of indigenous linear plasmids in actinobacteria, the engineering strategy described here could be implemented in a variety of strains for the overproduction of various natural products.

  6. Production of Ramoplanin and Ramoplanin Analogs by Actinomycetes.

    PubMed

    de la Cruz, Mercedes; González, Ignacio; Parish, Craig A; Onishi, Russell; Tormo, José R; Martín, Jesús; Peláez, Fernando; Zink, Debbie; El Aouad, Noureddine; Reyes, Fernando; Genilloud, Olga; Vicente, Francisca

    2017-01-01

    Ramoplanin is a glycolipodepsipeptide antibiotic obtained from fermentation of Actinoplanes sp. ATCC 33076 that exhibits activity against clinically important multi-drug-resistant, Gram-positive pathogens including vancomycin-resistant Enterococcus (VRE), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-intermediate resistant Clostridium difficile. It disrupts bacterial cell wall through a unique mechanism of action by sequestering the peptidoglycan intermediate Lipid II and therefore does not show cross-resistance with other antibiotics. However, while demonstrating excellent antimicrobial activity in systemic use in animal models of infection, ramoplanin presents low local tolerability when injected intravenously. As a consequence of this limitation, new derivatives are desirable to overcome this issue. During a natural product screening program developed to discover compounds that disrupt bacterial cell wall synthesis by inhibiting peptidoglycan transglycosylation through binding to the intermediate Lipid II, 49 actinomycete strains were identified by HR-LCMS as producers of ramoplanin-related compounds. The producing strains were isolated from environmental samples collected worldwide comprising both tropical and temperate areas. To assess the diversity of this microbial population, the 49 isolates were initially identified to the genus level on the basis of their micromorphology, and 16S sequencing confirmed the initial identification of the strains. These analyses resulted in the identification of members of genus Streptomyces, as well as representatives of the families Micromonosporaceae, Nocardiaceae, Thermomonosporaceae, and Pseudonocardiaceae, suggesting that the production of ramoplanins is relatively widespread among Actinomycetes. In addition, all of these isolates were tested against a panel of Gram-positive and Gram-negative bacteria, filamentous fungi, and yeast in order to further characterize their antimicrobial properties. This

  7. Production of Ramoplanin and Ramoplanin Analogs by Actinomycetes

    PubMed Central

    de la Cruz, Mercedes; González, Ignacio; Parish, Craig A.; Onishi, Russell; Tormo, José R.; Martín, Jesús; Peláez, Fernando; Zink, Debbie; El Aouad, Noureddine; Reyes, Fernando; Genilloud, Olga; Vicente, Francisca

    2017-01-01

    Ramoplanin is a glycolipodepsipeptide antibiotic obtained from fermentation of Actinoplanes sp. ATCC 33076 that exhibits activity against clinically important multi-drug-resistant, Gram-positive pathogens including vancomycin-resistant Enterococcus (VRE), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-intermediate resistant Clostridium difficile. It disrupts bacterial cell wall through a unique mechanism of action by sequestering the peptidoglycan intermediate Lipid II and therefore does not show cross-resistance with other antibiotics. However, while demonstrating excellent antimicrobial activity in systemic use in animal models of infection, ramoplanin presents low local tolerability when injected intravenously. As a consequence of this limitation, new derivatives are desirable to overcome this issue. During a natural product screening program developed to discover compounds that disrupt bacterial cell wall synthesis by inhibiting peptidoglycan transglycosylation through binding to the intermediate Lipid II, 49 actinomycete strains were identified by HR-LCMS as producers of ramoplanin-related compounds. The producing strains were isolated from environmental samples collected worldwide comprising both tropical and temperate areas. To assess the diversity of this microbial population, the 49 isolates were initially identified to the genus level on the basis of their micromorphology, and 16S sequencing confirmed the initial identification of the strains. These analyses resulted in the identification of members of genus Streptomyces, as well as representatives of the families Micromonosporaceae, Nocardiaceae, Thermomonosporaceae, and Pseudonocardiaceae, suggesting that the production of ramoplanins is relatively widespread among Actinomycetes. In addition, all of these isolates were tested against a panel of Gram-positive and Gram-negative bacteria, filamentous fungi, and yeast in order to further characterize their antimicrobial properties. This

  8. [Bioactivity of endophytic actinomycetes from medicinal plants and secondary metabolites from strain D62].

    PubMed

    Liu, Ning; Zhang, Hui; Zheng, Wen; Huang, Ying; Wang, Hai-Bin

    2007-10-01

    It is believed that genetic recombination of the endophytes with the hosts that occurred in evolutionary time could result in some endophytes producing certain phytochemical originally characteristic of the host. Based on this widely accepted hypothesis, there have been increasing research efforts focused on screening for novel natural products from endophytes. In this study, antimicrobial and antitumor activities of 165 actinomycetes isolated from medicinal plants collected from Xishuangbanna were tested by agar diffusion method and WST-8 assay respectively. The results showed that over 42% of the isolates exhibited antagonism against pathogenic strains, and 54.5% displayed excellent inhibition against mouse melanoma cell line B16 or/and human alveolar epithelial cell line A549. These results are superior to those of soil actinomycetes, indicating tremendous potential of endophytic of actinomycetes for exploration. Six compounds that had both antimicrobial and antitumor activities were separated and purified from isolate Streptomyces sp. D62 by resin adsorption, silica-gel column and sephadex chromatography, etc. On the basis of spectral analyses, they were identified as antimycin A4a (1), antimycin A7a (2), antimycin A2a (3), antimycin A1a (4), 10-hydroxy-10-methyl-dodec-2-en-1,4-olide (5) and 6-(2-(4-aminophenyl)-2-oxoethyl)-3,5-dimethyl-tetrahydropyran-2-one(6), with the last one defined as a novel compound. Based on all these results, it is convinced that endophytic actinomycetes are a promising resource for bioactive natural product discovery.

  9. Thioesterase Domain Swapping of a Linear Polyketide Tautomycetin with a Macrocyclic Polyketide Pikromycin in Streptomyces sp. CK4412

    PubMed Central

    Tripathi, Ashootosh; Choi, Si-Sun; Sherman, David H.; Kim, Eung-Soo

    2016-01-01

    Tautomycetin (TMC) is a linear polyketide metabolite produced by Streptomyces sp. CK4412 that has been reported to possess multiple biological functions including T cell-specific immunosuppressive and anticancer activities that occur through a mechanism of differential inhibition of protein phosphatases such as PP1, PP2A, and SHP2. We previously reported the characterization of the entire TMC biosynthetic gene cluster constituted by multifunctional type I polyketide synthase (PKS) assembly and suggested that the linear form of TMC could be generated via free acid chain termination by a narrow TMC thioesterase (TE) pocket. The modular nature of the assembly presents a unique opportunity to alter or interchange the native biosynthetic domains to produce targeted variants of TMC. Herein, we report swapping of the TMC TE domain sequence with the exact counterpart of the macrocyclic polyketide pikromycin (PIK) TE. PIK TE-swapped Streptomyces sp. CK4412 mutant produced not only TMC, but also a cyclized form of TMC, implying that the bioengineering based in vivo custom construct can be exploited to produce engineered macrolactones with new structural functionality. PMID:27277081

  10. Simultaneous chromate and sulfate removal by Streptomyces sp. MC1. Changes in intracellular protein profile induced by Cr(VI).

    PubMed

    Bonilla, José Oscar; Callegari, Eduardo Alberto; Delfini, Claudio Daniel; Estevez, María Cristina; Villegas, Liliana Beatriz

    2016-11-01

    The purpose of this study was to investigate the influence of increasing sulfate concentrations on chromium removal, to evaluate the effect of the presence of Cr(VI) on sulfate removal by Streptomyces sp. MC1 and to analyze the differential protein expression profile in the presence of this metal for the identification of proteins repressed or overexpressed. In the presence of Cr(VI) but in the absence of sulfate ions, bacterial growth was negligible, showing the Cr(VI) toxicity for this bacterium. However, the sulfate presence stimulated bacterium growth and Cr(VI) removal, regardless of its concentrations. Streptomyces sp. MC1 showed ability to remove chromium and sulfate simultaneously. Also, the sulfate presence favored the decrease of total chromium concentration from supernatants reaching a decrease of 50% at 48 h. In presence of chromium, seven proteins were down-expressed and showed homology to proteins involved in protein biosynthesis, energy production and free radicals detoxification while two proteins involved in oxidation-reduction processes identified as dihydrolipoamide dehydrogenase and S-adenosyl-l-methionine synthase were overexpressed.

  11. Diversity of ABBA Prenyltransferases in Marine Streptomyces sp. CNQ-509: Promiscuous Enzymes for the Biosynthesis of Mixed Terpenoid Compounds

    PubMed Central

    Leipoldt, Franziska; Zeyhle, Philipp; Kulik, Andreas; Kalinowski, Jörn; Heide, Lutz; Kaysser, Leonard

    2015-01-01

    Terpenoids are arguably the largest and most diverse family of natural products, featuring prominently in e.g. signalling, self-defence, UV-protection and electron transfer. Prenyltransferases are essential players in terpenoid and hybrid isoprenoid biosynthesis that install isoprene units on target molecules and thereby often modulate their bioactivity. In our search for new prenyltransferase biocatalysts we focused on the marine-derived Streptomyces sp. CNQ-509, a particularly rich source of meroterpenoid chemistry. Sequencing and analysis of the genome of Streptomyces sp. CNQ-509 revealed seven putative phenol/phenazine-specific ABBA prenyltransferases, and one putative indole-specific ABBA prenyltransferase. To elucidate the substrate specificity of the ABBA prenyltransferases and to learn about their role in secondary metabolism, CnqP1 –CnqP8 were produced in Escherichia coli and incubated with various aromatic and isoprenoid substrates. Five of the eight prenyltransferases displayed enzymatic activity. The efficient conversion of dihydroxynaphthalene derivatives by CnqP3 (encoded by AA958_24325) and the co-location of AA958_24325 with genes characteristic for the biosynthesis of THN (tetrahydroxynaphthalene)-derived natural products indicates that the enzyme is involved in the formation of debromomarinone or other naphthoquinone-derived meroterpenoids. Moreover, CnqP3 showed high flexibility towards a range of aromatic and isoprenoid substrates and thus represents an interesting new tool for biocatalytic applications. PMID:26659564

  12. In vitro probiotic profile based selection of indigenous actinobacterial probiont Streptomyces sp. JD9 for enhanced broiler production.

    PubMed

    Latha, Selvanathan; Vinothini, Gopal; John Dickson Calvin, Devadasan; Dhanasekaran, Dharumadurai

    2016-01-01

    The present study was undertaken to select exclusive indigenous actinobacterial probiont for broiler health improvement based on in vitro probiotic potentials. In total, 18 actinobacterial cultures isolated from chicken were screened for survivability (resistance to low pH, pepsin, bile and pancreatin), colonization (auto-aggregation, hydrophobicity and co-aggregation) and safety (antibiotic susceptibility and non-haemolytic activity). Ten cultures showed excellent viability at pH 2 and most of the acid-tolerant isolates exhibited resistance to pepsin (3 mg/mL), bile (0.3%) and pancreatin (1 mg/mL). Besides, the examined isolates displayed efficient adhesion properties. All the isolates were susceptible to 9 different antibiotics and none of them exhibited β-haemolytic activity. Moreover, the culture JD9 revealed remarkable probiotic features compared to the other isolates, which was identified as Streptomyces sp. JD9 (KF878075). Taken together, the present study suggests that the probiont Streptomyces sp. JD9 could potentially be used in broiler practices as a feed additive to facilitate enhanced broiler production.

  13. Characterization of the Streptomyces sp. Strain C5 snp Locus and Development of snp-Derived Expression Vectors

    PubMed Central

    DeSanti, Charles L.; Strohl, William R.

    2003-01-01

    The Streptomyces sp. strain C5 snp locus is comprised of two divergently oriented genes: snpA, a metalloproteinase gene, and snpR, which encodes a LysR-like activator of snpA transcription. The transcriptional start point of snpR is immediately downstream of a strong T-N11-A inverted repeat motif likely to be the SnpR binding site, while the snpA transcriptional start site overlaps the ATG start codon, generating a leaderless snpA transcript. By using the aphII reporter gene of pIJ486 as a reporter, the plasmid-borne snpR-activated snpA promoter was ca. 60-fold more active than either the nonactivated snpA promoter or the melC1 promoter of pIJ702. The snpR-activated snpA promoter produced reporter protein levels comparable to those of the up-mutated ermE∗ promoter. The SnpR-activated snpA promoter was built into a set of transcriptional and translational fusion expression vectors which have been used for the intracellular expression of numerous daunomycin biosynthesis pathway genes from Streptomyces sp. strain C5 as well as the expression and secretion of soluble recombinant human endostatin. PMID:12620855

  14. A fibrinolytic protease AfeE from Streptomyces sp. CC5, with potent thrombolytic activity in a mouse model.

    PubMed

    Sun, Zhibin; Liu, Pingping; Cheng, Guangyan; Zhang, Biying; Dong, Weiliang; Su, Xingli; Huang, Yan; Cui, Zhongli; Kong, Yi

    2016-04-01

    Fibrinolytic proteases have potential applications in cardiovascular disease therapy. A novel fibrinolytic protease, AfeE, with strong thrombolytic activity was purified from Streptomyces sp. CC5. AfeE displayed maximum activity at 40°C in the pH range of 7.0-12.0. It was strongly inhibited by serine protease inhibitor phenylmethanesulfonylfluoride, soybean trypsin inhibitor, tosyl-l-lysine chloromethyl ketone and tosyl-l-phenylalanine chloromethyl ketone. The activity of the enzyme was partially inhibited by Cu(2+), Co(2+) and Zn(2+). AfeE exhibited higher substrate specificity for fibrin than fibrinogen, which has rarely been reported in fibrinolytic enzymes. AfeE also showed high thrombolytic activity in a carrageenan-induced mouse tail thrombosis model. AfeE prolonged prothrombin time, activated partial thromboplastin time, and thrombin time in rat blood. A bleeding time assay revealed that AfeE did not prolong bleeding time in mice at a dose of 1mg/kg. No acute cytotoxicity was observed for AfeE at 320μg/well in human umbilical vein endothelial cells. The afeE gene was cloned from the genome of Streptomyces sp. CC5. Full-length AFE-CC5E contained 434 amino acids and was processed into a mature form consisting 284 amino acids by posttranslational modification, as revealed by high-resolution mass spectrometry analysis. These results indicate that AfeE is a prospective candidate for antithrombotic drug development.

  15. Imaging secondary metabolism of Streptomyces sp. Mg1 during cellular lysis and colony degradation of competing Bacillus subtilis.

    PubMed

    Barger, Sarah R; Hoefler, B Chris; Cubillos-Ruiz, Andrés; Russell, William K; Russell, David H; Straight, Paul D

    2012-10-01

    Soil streptomycetes are saprotrophic bacteria that secrete numerous secondary metabolites and enzymes for extracellular functions. Many streptomycetes produce antibiotics thought to protect vegetative mycelia from competing organisms. Here we report that an organism isolated from soil, Streptomyces sp. Mg1, actively degrades colonies and causes cellular lysis of Bacillus subtilis when the organisms are cultured together. We predicted that the inhibition and degradation of B. subtilis colonies in this competition depends upon a combination of secreted factors, including small molecule metabolites and enzymes. To begin to unravel this complex competitive phenomenon, we use a MALDI imaging mass spectrometry strategy to map the positions of metabolites secreted by both organisms. In this report, we show that Streptomyces sp. Mg1 produces the macrolide antibiotic chalcomycin A, which contributes to inhibition of B. subtilis growth in combination with other, as yet unidentified factors. We suggest that efforts to understand competitive and cooperative interactions between bacterial species benefit from assays that pair living organisms and probe the complexity of metabolic exchanges between them.

  16. Detection and identification of novel actinomycetes.

    PubMed

    Williams, S T; Locci, R; Beswick, A; Kurtböke, D I; Kuznetsov, V D; Le Monnier, F J; Long, P F; Maycroft, K A; Palma, R A; Petrolini, B

    1993-10-01

    The actinomycetes are well known as a group of filamentous, Gram-positive bacteria that produce many useful secondary metabolites, including antibiotics and enzymes. Although they have been intensively studied for both theoretical and practical objectives, there is much scope for developing our basic knowledge of the means of detection and isolation of these microbes. This session concentrated on new methods for the detection and identification of novel actinomycetes from a range of environments. Approaches to the detection of actinomycetes ranged from investigations of neglected habitats and extreme environments (e.g. alkaline soils and oil drills) to the analysis of DNA extracted from the environment and use of specific phages. The continuing problems of the identification of actinomycete isolates were also considered. Topics discussed included use of phage typing, DNA probes, and correlation between phenetic and genotypic species of Streptomyces.

  17. Role of Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase in Oleaginous Streptomyces sp. Strain G25

    PubMed Central

    Röttig, Annika; Strittmatter, Carl Simon; Schauer, Jennifer; Hiessl, Sebastian; Daniel, Rolf

    2016-01-01

    ABSTRACT Recently, we isolated a novel Streptomyces strain which can accumulate extraordinarily large amounts of triacylglycerol (TAG) and consists of 64% fatty acids (dry weight) when cultivated with glucose and 50% fatty acids (dry weight) when cultivated with cellobiose. To identify putative gene products responsible for lipid storage and cellobiose utilization, we analyzed its draft genome sequence. A single gene encoding a wax ester synthase/acyl coenzyme A (CoA):diacylglycerol acyltransferase (WS/DGAT) was identified and heterologously expressed in Escherichia coli. The purified enzyme AtfG25 showed acyltransferase activity with C12- or C16-acyl-CoA, C12 to C18 alcohols, or dipalmitoyl glycerol. This acyltransferase exhibits 24% amino acid identity to the model enzyme AtfA from Acinetobacter baylyi but has high sequence similarities to WS/DGATs from other Streptomyces species. To investigate the impact of AtfG25 on lipid accumulation, the respective gene, atfG25, was inactivated in Streptomyces sp. strain G25. However, cells of the insertion mutant still exhibited DGAT activity and were able to store TAG, albeit in lower quantities and at lower rates than the wild-type strain. These findings clearly indicate that AtfG25 has an important, but not exclusive, role in TAG biosynthesis in the novel Streptomyces isolate and suggest the presence of alternative metabolic pathways for lipid accumulation which are discussed in the present study. IMPORTANCE A novel Streptomyces strain was isolated from desert soil, which represents an extreme environment with high temperatures, frequent drought, and nutrient scarcity. We believe that these harsh conditions promoted the development of the capacity for this strain to accumulate extraordinarily large amounts of lipids. In this study, we present the analysis of its draft genome sequence with a special focus on enzymes potentially involved in its lipid storage. Furthermore, the activity and importance of the detected

  18. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    DOE PAGES

    Köberl, Martina; White, Richard A.; Erschen, Sabine; ...

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria, and nematodes. The 8.2-Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  19. Draft Genome Sequence of Streptomyces sp. AVP053U2 Isolated from Styela clava, a Tunicate Collected in Long Island Sound

    PubMed Central

    deMayo, James A.; Maas, Kendra R.

    2016-01-01

    Streptomyces sp. AVP053U2 is a marine bacterium isolated from Styela clava, a tunicate collected in Long Island Sound. Here, we report a draft genome for this bacterium, which was found to contain a high capacity for secondary metabolite production based on analysis and identification of numerous biosynthetic gene clusters. PMID:27738023

  20. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    SciTech Connect

    Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria and nematodes. The 8.2 Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  1. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    SciTech Connect

    Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria, and nematodes. The 8.2-Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  2. Genomic and Secondary Metabolite Analyses of Streptomyces sp. 2AW Provide Insight into the Evolution of the Cycloheximide Pathway

    PubMed Central

    Stulberg, Elizabeth R.; Lozano, Gabriel L.; Morin, Jesse B.; Park, Hyunjun; Baraban, Ezra G.; Mlot, Christine; Heffelfinger, Christopher; Phillips, Gillian M.; Rush, Jason S.; Phillips, Andrew J.; Broderick, Nichole A.; Thomas, Michael G.; Stabb, Eric V.; Handelsman, Jo

    2016-01-01

    The dearth of new antibiotics in the face of widespread antimicrobial resistance makes developing innovative strategies for discovering new antibiotics critical for the future management of infectious disease. Understanding the genetics and evolution of antibiotic producers will help guide the discovery and bioengineering of novel antibiotics. We discovered an isolate in Alaskan boreal forest soil that had broad antimicrobial activity. We elucidated the corresponding antimicrobial natural products and sequenced the genome of this isolate, designated Streptomyces sp. 2AW. This strain illustrates the chemical virtuosity typical of the Streptomyces genus, producing cycloheximide as well as two other biosynthetically unrelated antibiotics, neutramycin, and hygromycin A. Combining bioinformatic and chemical analyses, we identified the gene clusters responsible for antibiotic production. Interestingly, 2AW appears dissimilar from other cycloheximide producers in that the gene encoding the polyketide synthase resides on a separate part of the chromosome from the genes responsible for tailoring cycloheximide-specific modifications. This gene arrangement and our phylogenetic analyses of the gene products suggest that 2AW holds an evolutionarily ancestral lineage of the cycloheximide pathway. Our analyses support the hypothesis that the 2AW glutaramide gene cluster is basal to the lineage wherein cycloheximide production diverged from other glutarimide antibiotics. This study illustrates the power of combining modern biochemical and genomic analyses to gain insight into the evolution of antibiotic-producing microorganisms. PMID:27199910

  3. Genomic and Secondary Metabolite Analyses of Streptomyces sp. 2AW Provide Insight into the Evolution of the Cycloheximide Pathway.

    PubMed

    Stulberg, Elizabeth R; Lozano, Gabriel L; Morin, Jesse B; Park, Hyunjun; Baraban, Ezra G; Mlot, Christine; Heffelfinger, Christopher; Phillips, Gillian M; Rush, Jason S; Phillips, Andrew J; Broderick, Nichole A; Thomas, Michael G; Stabb, Eric V; Handelsman, Jo

    2016-01-01

    The dearth of new antibiotics in the face of widespread antimicrobial resistance makes developing innovative strategies for discovering new antibiotics critical for the future management of infectious disease. Understanding the genetics and evolution of antibiotic producers will help guide the discovery and bioengineering of novel antibiotics. We discovered an isolate in Alaskan boreal forest soil that had broad antimicrobial activity. We elucidated the corresponding antimicrobial natural products and sequenced the genome of this isolate, designated Streptomyces sp. 2AW. This strain illustrates the chemical virtuosity typical of the Streptomyces genus, producing cycloheximide as well as two other biosynthetically unrelated antibiotics, neutramycin, and hygromycin A. Combining bioinformatic and chemical analyses, we identified the gene clusters responsible for antibiotic production. Interestingly, 2AW appears dissimilar from other cycloheximide producers in that the gene encoding the polyketide synthase resides on a separate part of the chromosome from the genes responsible for tailoring cycloheximide-specific modifications. This gene arrangement and our phylogenetic analyses of the gene products suggest that 2AW holds an evolutionarily ancestral lineage of the cycloheximide pathway. Our analyses support the hypothesis that the 2AW glutaramide gene cluster is basal to the lineage wherein cycloheximide production diverged from other glutarimide antibiotics. This study illustrates the power of combining modern biochemical and genomic analyses to gain insight into the evolution of antibiotic-producing microorganisms.

  4. TMG-chitotriomycin, an enzyme inhibitor specific for insect and fungal beta-N-acetylglucosaminidases, produced by actinomycete Streptomyces anulatus NBRC 13369.

    PubMed

    Usuki, Hirokazu; Nitoda, Teruhiko; Ichikawa, Misato; Yamaji, Nahoko; Iwashita, Takashi; Komura, Hajime; Kanzaki, Hiroshi

    2008-03-26

    A novel beta-N-acetylglucosaminidase (GlcNAcase) inhibitor named TMG-chitotriomycin (1) was isolated from the culture filtrate of Streptomyces anulatus NBRC13369. The strain produced 1 only when colloidal chitin was used as the sole carbon source in the production medium. The structure of 1 was determined by spectral and constitutive sugar analyses of the corresponding alditol derivatives to be an equilibrated mixture of alpha-d-N,N,N-triMeGlcNH2-(1,4)-beta-d-GlcNAc-(1,4)-beta-d-GlcNAc-(1,4)-d-GlcNAc and its C-2 epimer of the reducing end residue. TMG-chitotriomycin (1) showed potent and selective inhibition of insect and fungal GlcNAcases with no inhibition of mammalian and plant GlcNAcases. In contrast, the known GlcNAcase inhibitor nagstatin potently inhibited all GlcNAcases. It should be emphasized that synthesized d-N,N,N-triMeGlcNH2, which is the component sugar of 1, showed no inhibition of the insect Spodoptera litura GlcNAcase. These results suggest that the (GlcNAc)3 unit positioned at the reducing end of 1 is essential for its enzyme inhibitory activity. The unique inhibitory spectrum of 1 will be useful to study chitinolytic systems and to develop selective fungicides or pesticides.

  5. Oligotrophy is Helpful for the Isolation of Bioactive Actinomycetes.

    PubMed

    Wang, Dong-Sheng; Xue, Quan-Hong; Ma, Yun-Yan; Wei, Xiao-Li; Chen, Jie; He, Fei

    2014-06-01

    It is necessary to develop new methods for the isolation of unknown actinomycetes from soils. To evaluate the effects of oligotrophic medium on the isolation of soil actinomycetes and develop a new isolation method, the Gause's synthetic medium was diluted to one tenth the recommended concentration in the present study. Soil dilution plate technique was used to isolate actinomycetes from the soil samples. Oligotrophy decreased actinomycete and streptomycete counts, as well as the number of antagonistic actinomycete species. Oligotrophy also decreased the number of actinomycete species in five samples. Some actinomycete species were cultured only on the oligotrophic medium, whereas other species could not be cultured. Oligotrophy decreased actinomycete counts more significantly for soils with organic matter content >40 g/kg. We used 16S rRNA sequence analysis to identify 22 actinomycete species that were only cultured on the oligotrophic medium. Oligotrophic medium was helpful for the isolation of Streptomyces spp., Micromonospora spp. and Streptosporangium spp. Slightly more than 80 % of the identified actinomycete species were biologically active. Therefore, we could draw a conclusion that oligotrophic medium could be helpful for the discovery of new antibiotic producers and the exploitation and utilization of new, biologically active compounds.

  6. Metabolomics-Driven Discovery of a Prenylated Isatin Antibiotic Produced by Streptomyces Species MBT28.

    PubMed

    Wu, Changsheng; Du, Chao; Gubbens, Jacob; Choi, Young Hae; van Wezel, Gilles P

    2015-10-23

    Actinomycetes are a major source of antimicrobials, anticancer compounds, and other medically important products, and their genomes harbor extensive biosynthetic potential. Major challenges in the screening of these microorganisms are to activate the expression of cryptic biosynthetic gene clusters and the development of technologies for efficient dereplication of known molecules. Here we report the identification of a previously unidentified isatin-type antibiotic produced by Streptomyces sp. MBT28, following a strategy based on NMR-based metabolomics combined with the introduction of streptomycin resistance in the producer strain. NMR-guided isolation by tracking the target proton signal resulted in the characterization of 7-prenylisatin (1) with antimicrobial activity against Bacillus subtilis. The metabolite-guided genome mining of Streptomyces sp. MBT28 combined with proteomics identified a gene cluster with an indole prenyltransferase that catalyzes the conversion of tryptophan into 7-prenylisatin. This study underlines the applicability of NMR-based metabolomics in facilitating the discovery of novel antibiotics.

  7. Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential

    PubMed Central

    Passari, Ajit K.; Mishra, Vineet K.; Saikia, Ratul; Gupta, Vijai K.; Singh, Bhim P.

    2015-01-01

    Microorganisms associated with medicinal plants are of interest as the producers of important bioactive compounds. To date, the diversity of culturable endophytic actinomycetes associated with medicinal plants is in its initial phase of exploration. In this study, 42 endophytic actinomycetes were isolated from different organs of seven selected medicinal plants. The highest number of isolates (n = 22, 52.3%) of actinomycetes was isolated from roots, followed by stems (n = 9, 21.4%), leaves (n = 6, 14.2%), flowers (n = 3, 7.1%), and petioles (n = 2, 4.7%). The genus Streptomyces was the most dominant among the isolates (66.6%) in both the locations (Dampa TRF and Phawngpuii NP, Mizoram, India). From a total of 42 isolates, 22 isolates were selected for further studies based on their ability to inhibit one of the tested human bacterial or fungal pathogen. Selected isolates were identified based on 16S rRNA gene analysis and subsequently the isolates were grouped to four different genera; Streptomyces, Brevibacterium, Microbacterium, and Leifsonia. Antibiotic sensitivity assay was performed to understand the responsible antimicrobials present in the isolates showing the antimicrobial activities and revealed that the isolates were mostly resistant to penicillin G and ampicillin. Further, antimicrobial properties and antibiotic sensitivity assay in combination with the results of amplification of biosynthetic genes polyketide synthase (PKS-I) and non-ribosomal peptide synthetase (NRPS) showed that the endophytic actinomycetes associated with the selected medicinal plants have broad-spectrum antimicrobial activity. This is the first report of the isolation of Brevibacterium sp., Microbacterium sp., and Leifsonia xyli from endophytic environments of medicinal plants, Mirabilis jalapa and Clerodendrum colebrookianum. Our results emphasize that endophytic actinomycetes associated with medicinal plants are an unexplored resource for the discovery of biologically active

  8. [Biosynthetic study of actinomycetes-metabolites for creating novel analogs].

    PubMed

    Ito, Takuya

    2013-01-01

    The aminocyclitol family is a relatively new class of natural products such as gentamicin, kanamycin, and streptomycin, which have been used clinically for decades as potent antimicrobial agents. These secondary metabolites are chiefly produced by microorganisms, especially Actinomycetes. Their chemical structures most commonly contain a C7N unit, 2-epi-5-epi-valiolone or 3-amino-5-hydroxybenzoic acid (3,5-AHBA) which are known to be responsible for their biological activities. In the course of current study, the biosynthesis of the C7N-containing metabolites, validamycin and acarbose, pactamycin, have been evaluated. We studied N-formamide salicylic acid (FSA) moiety which is a C7N unit synthesized from tryptophan by microorganisms. A strong antifungal agent antimycin, isolated from several Streptomyces sp., contains an FSA moiety, and constitutes a unique nine-membered dilactone ring with L-threonine, short-chain fatty acid, and an amide linkage connecting it to an FSA moiety. Also, an antitumor antibiotic asukamycin, produced by Streptomyces nodosus subsp. asukaensis ATCC 29757, consists of both 3,4-AHBA and C5N, cyclohexane ring linked to trans-triens. To improve the efficacy and reduce the toxicity of these metabolites, further structural modification is needed. Total chemical synthesis of these complex compounds is difficult. Therefore, alternative approaches are required, e.g., biosynthetic or genetic modification methods. This review presents the biosynthetic study on these compounds for creating new analogs using mutasyntheis.

  9. Pseudonocardians A–C, New Diazaanthraquinone Derivatives from a Deap-Sea Actinomycete Pseudonocardia sp. SCSIO 01299

    PubMed Central

    Li, Sumei; Tian, Xinpeng; Niu, Siwen; Zhang, Wenjun; Chen, Yuchan; Zhang, Haibo; Yang, Xianwen; Zhang, Weimin; Li, Wenjun; Zhang, Si; Ju, Jianhua; Zhang, Changsheng

    2011-01-01

    Pseudonocardians A–C (2–4), three new diazaanthraquinone derivatives, along with a previously synthesized compound deoxynyboquinone (1), were produced by the strain SCSIO 01299, a marine actinomycete member of the genus Pseudonocardia, isolated from deep-sea sediment of the South China Sea. The structures of compounds 1–4 were determined by mass spectrometry and NMR experiments (1H, 13C, HSQC, and HMBC). The structure of compound 1, which was obtained for the first time from a natural source, was confirmed by X-ray analysis. Compounds 1–3 exhibited potent cytotoxic activities against three tumor cell lines of SF-268, MCF-7 and NCI-H460 with IC50 values between 0.01 and 0.21 μm, and also showed antibacterial activities on Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212 and Bacillus thuringensis SCSIO BT01, with MIC values of 1–4 μg mL−1. PMID:21892356

  10. Investigation of Antioxidative and Anticancer Potentials of Streptomyces sp. MUM256 Isolated from Malaysia Mangrove Soil

    PubMed Central

    Tan, Loh Teng-Hern; Ser, Hooi-Leng; Yin, Wai-Fong; Chan, Kok-Gan; Lee, Learn-Han; Goh, Bey-Hing

    2015-01-01

    A Streptomyces strain, MUM256 was isolated from Tanjung Lumpur mangrove soil in Malaysia. Characterization of the strain showed that it has properties consistent with those of the members of the genus Streptomyces. In order to explore the potential bioactivities, extract of the fermented broth culture of MUM256 was prepared with organic solvent extraction method. DPPH and SOD activity were utilized to examine the antioxidant capacity and the results have revealed the potency of MUM256 in superoxide anion scavenging activity in dose-dependent manner. The cytotoxicity of MUM256 extract was determined using cell viability assay against 8 different panels of human cancer cell lines. Among all the tested cancer cells, HCT116 was the most sensitive toward the extract treatment. At the highest concentration of tested extract, the result showed 2.3-, 2.0-, and 1.8-folds higher inhibitory effect against HCT116, HT29, and Caco-2 respectively when compared to normal cell line. This result has demonstrated that MUM256 extract was selectively cytotoxic toward colon cancer cell lines. In order to determine the constituents responsible for its bioactivities, the extract was then subjected to chemical analysis using GC-MS. The analysis resulted in the identification of chemical constituents including phenolic and pyrrolopyrazine compounds which may responsible for antioxidant and anticancer activities observed. Based on the findings of this study, the presence of bioactive constituents in MUM256 extract could be a potential source for the development of antioxidative and chemopreventive agents. PMID:26635777

  11. Two new isoflavone 7-O-α-4″-anhydro-4″,5″-didehydroglucuronides from Streptomyces sp. LZ35ΔgdmAI.

    PubMed

    Deng, Jing-Jing; Lu, Chun-Hua

    2016-01-01

    Two isoflavone 7-O-α-4″-anhydro-4″,5″-didehydroglucuronides, namely daidzein 7-O-α-4″-anhydro-4″,5″-didehydroglucuronide (1) and genistein 7-O-α-4″-anhydro-4″,5″-didehydroglucuronide (2), were isolated and identified from the mutant strain of Streptomyces sp. LZ35ΔgdmAI. Their structures were elucidated by the analysis of their high resolution mass spectrometry (HR-MS) and 1D, 2D Nuclear magnetic Resonance (NMR) spectroscopic data. They are new natural products and maybe the transformed products of the soybean meal by Streptomyces sp. LZ35ΔgdmAI.

  12. Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease- and SDS-resistant xylanase.

    PubMed

    Zhou, Junpei; Shi, Pengjun; Zhang, Rui; Huang, Huoqing; Meng, Kun; Yang, Peilong; Yao, Bin

    2011-04-01

    A pH-stable and protease-resistant xylanase (XynB119) was identified from Streptomyces sp. TN119, a strain isolated from the gut luminal contents of longhorned beetle (Batocera horsfieldi) larvae. Using the GC TAIL-PCR method, the 1,026-bp coding gene (xynB119) with 67.3% GC content was successfully cloned and expressed in Escherichia coli. It encodes a 341-residue polypeptide with a calculated molecular mass of 35.9 kDa, including a putative 41-residue signal peptide, a catalytic domain of glycosyl hydrolase (GH) family 11, a short Gly/Pro-rich linker, and a family 2 cellulose-binding domain (CBM 2). The deduced amino acid sequence is most similar to (61.9% identity) an endo-1,4-β-xylanase from Streptomyces thermoviolaceus OPC-520. Purified recombinant XynB119 exhibited peak activity at 50 °C and pH 7.0, remained stable over a broad pH range (retaining >70% activity after incubation at pH 1.0-11.0 for 1 h at 37 °C without substrate), had strong protease resistance (retaining >90% activity after proteolytic treatment at 37 °C for 1 h) and SDS resistance (at 100 mM). These properties make XynB119 promising for application in the feed industry and valuable for basic research. Compared to r-XynB119, the r-XynB119 derivative without CBM 2 and linker region (r-XynB119d) exhibited a decreased pH stability of >25% at extreme pHs (pH 1.0-3.0 and pH 11.0-12.0).

  13. Purification and biochemical characterization of two detergent-stable serine alkaline proteases from Streptomyces sp. strain AH4.

    PubMed

    Touioui, Souraya Boulkour; Jaouadi, Nadia Zaraî; Boudjella, Hadjira; Ferradji, Fatma Zohra; Belhoul, Mouna; Rekik, Hatem; Badis, Abdelmalek; Bejar, Samir; Jaouadi, Bassem

    2015-07-01

    Streptomyces sp. strain AH4 exhibited a high ability to produce two extracellular proteases when cultured on a yeast malt-extract (ISP2)-casein-based medium. Pure proteins were obtained after heat treatment (30 min at 70 °C) and ammonium sulphate fractionation (30-60 %), followed by size exclusion HPLC column. Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis revealed that the purified enzymes (named SAPS-P1 and SAPS-P2) were monomers with molecular masses of 36,417.13 and 21,099.10 Da, respectively. Their identified N-terminal amino acid displayed high homologies with those of Streptomyces proteases. While SAPS-P1 was optimally active at pH 12.0 and 70 °C, SAPS-P2 showed optimum activity at pH 10.0 and 60 °C. Both enzymes were completely stable within a wide range of temperature (45-75 °C) and pH (8.0-11.5). They were noted to be completely inhibited by phenylmethanesulfonyl fluoride and diisopropyl fluorophosphates, which confirmed their belonging to the serine proteases family. Compared to SAPS-P2, SAPS-P1 showed high thermostability and excellent stability towards bleaching, denaturing, and oxidizing agents. Both enzymes displayed marked stability and compatibility with a wide range of commercial laundry detergents and significant catalytic efficiencies compared to Subtilisin Carlsberg and Protease SG-XIV. Overall, the results indicated that SAPS-P1 and SAPS-P2 can be considered as potential promising candidates for future application as bioadditives in detergent formulations.

  14. Microbispora sp. LGMB259 Endophytic Actinomycete Isolated from Vochysia divergens (Pantanal, Brazil) Producing β-Carbolines and Indoles with Biological Activity

    PubMed Central

    Savi, Daiani C.; Shaaban, Khaled A.; Vargas, Nathalia; Ponomareva, Larissa V.; Possiede, Yvelise M.; Thorson, Jon S.; Glienke, Chirlei; Rohr, Jürgen

    2014-01-01

    Endophytic actinomycetes encompass bacterial groups that are well known for the production of a diverse range of secondary metabolites. Vochysia divergens is a medicinal plant, common in the “Pantanal” region (Brazil) and was focus of many investigations, but never regarding its community of endophytic symbionts. During a screening program, an endophytic strain isolated from the V. divergens, was investigated for its potential to show biological activity. The strain was characterized as Microbispora sp. LGMB259 by spore morphology and molecular analyze using nucleotide sequence of the 16S rRNA gene. Strain LGMB259 was cultivated in R5A medium producing metabolites with significant antibacterial activity. The strain produced 4 chemically related β-carbolines, and 3 Indoles. Compound 1-Vinyl-β-carboline-3-carboxylic acid displayed potent activity against the Gram-positive bacterial strains Micrococcus luteus NRRL B-2618 and Kocuria rosea B-1106, and was highly active against two human cancer cell lines, namely the prostate cancer cell line PC3 and the non-small-cell lung carcinoma cell line A549, with IC50 values of 9.45 and 24.67 µM, respectively. 1-Vinyl-β-carboline-3-carboxylic acid also showed moderate activity against the yeast Saccharomyces cerevisiae ATCC204508, as well as the phytopathogenic fungiPhyllosticta citricarpa LGMB06 and Colletotrichum gloeosporioides FDC83. PMID:25385358

  15. A NADPH-dependent (S)-imine reductase (SIR) from Streptomyces sp. GF3546 for asymmetric synthesis of optically active amines: purification, characterization, gene cloning, and expression.

    PubMed

    Mitsukura, Koichi; Kuramoto, Tatsuya; Yoshida, Toyokazu; Kimoto, Norihiro; Yamamoto, Hiroaki; Nagasawa, Toru

    2013-09-01

    A NADPH-dependent (S)-imine reductase (SIR) was purified to be homogeneous from the cell-free extract of Streptomyces sp. GF3546. SIR appeared to be a homodimer protein with subunits of 30.5 kDa based on SDS-polyacrylamide gel electrophoresis and HPLC gel filtration. It also catalyzed the (S)-enantioselective reduction of not only 2-methyl-1-pyrroline (2-MPN) but also 1-methyl-3,4-dihydroisoquinoline and 6,7-dimethoxy-1-methyl-3,4-dihydroisoquinoline. Specific activities for their imines were 130, 44, and 2.6 nmol min(-1) mg(-1), and their optical purities were 92.7 % ee, 96.4 % ee, and >99 % ee, respectively. Using a NADPH-regenerating system, 10 mM 2-MPN was converted to amine with 100 % conversion and 92 % ee after 24 h. The amino acid sequence analysis revealed that SIR showed about 60 % identity to 6-phosphogluconate dehydrogenase. However, it showed only 37 % identity with Streptomyces sp. GF3587 (R)-imine reductase. Expression of SIR in Escherichia coli was achieved, and specific activity of the cell-free extract was about two times higher than that of the cell-free extract of Streptomyces sp. GF3546.

  16. ­Genomic data mining of the marine actinobacteria Streptomyces sp. H-KF8 unveils insights into multi-stress related genes and metabolic pathways involved in antimicrobial synthesis

    PubMed Central

    Undabarrena, Agustina; Ugalde, Juan A.; Seeger, Michael

    2017-01-01

    Streptomyces sp. H-KF8 is an actinobacterial strain isolated from marine sediments of a Chilean Patagonian fjord. Morphological characterization together with antibacterial activity was assessed in various culture media, revealing a carbon-source dependent activity mainly against Gram-positive bacteria (S. aureus and L. monocytogenes). Genome mining of this antibacterial-producing bacterium revealed the presence of 26 biosynthetic gene clusters (BGCs) for secondary metabolites, where among them, 81% have low similarities with known BGCs. In addition, a genomic search in Streptomyces sp. H-KF8 unveiled the presence of a wide variety of genetic determinants related to heavy metal resistance (49 genes), oxidative stress (69 genes) and antibiotic resistance (97 genes). This study revealed that the marine-derived Streptomyces sp. H-KF8 bacterium has the capability to tolerate a diverse set of heavy metals such as copper, cobalt, mercury, chromate and nickel; as well as the highly toxic tellurite, a feature first time described for Streptomyces. In addition, Streptomyces sp. H-KF8 possesses a major resistance towards oxidative stress, in comparison to the soil reference strain Streptomyces violaceoruber A3(2). Moreover, Streptomyces sp. H-KF8 showed resistance to 88% of the antibiotics tested, indicating overall, a strong response to several abiotic stressors. The combination of these biological traits confirms the metabolic versatility of Streptomyces sp. H-KF8, a genetically well-prepared microorganism with the ability to confront the dynamics of the fjord-unique marine environment. PMID:28229018

  17. MDN-0170, a New Napyradiomycin from Streptomyces sp. Strain CA-271078.

    PubMed

    Lacret, Rodney; Pérez-Victoria, Ignacio; Oves-Costales, Daniel; de la Cruz, Mercedes; Domingo, Elizabeth; Martín, Jesús; Díaz, Caridad; Vicente, Francisca; Genilloud, Olga; Reyes, Fernando

    2016-10-18

    A new napyradiomycin, MDN-0170 (1), was isolated from the culture broth of the marine-derived actinomycete strain CA-271078, together with three known related compounds identified as 4-dehydro-4a-dechloronapyradiomycin A1 (2), napyradiomycin A1 (3) and 3-chloro-6,8-dihydroxy-8-α-lapachone (4). The structure of the new compound was determined using a combination of spectroscopic techniques, including 1D and 2D NMR and electrospray-time of flight mass spectrometry (ESI-TOF MS). The relative configuration of compound 1, which contains two independent stereoclusters, has been established by molecular modelling in combination with nOe and coupling constant analyses. Biosynthetic arguments also allowed us to propose its absolute stereochemistry. The antimicrobial properties of the compounds isolated were evaluated against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Aspergillus fumigatus, and Candida albicans. The potent bioactivity previously reported for compounds 2 and 3 against methicillin-sensitive S. aureus has been extended to methicillin-resistant strains in this report.

  18. MDN-0170, a New Napyradiomycin from Streptomyces sp. Strain CA-271078

    PubMed Central

    Lacret, Rodney; Pérez-Victoria, Ignacio; Oves-Costales, Daniel; de la Cruz, Mercedes; Domingo, Elizabeth; Martín, Jesús; Díaz, Caridad; Vicente, Francisca; Genilloud, Olga; Reyes, Fernando

    2016-01-01

    A new napyradiomycin, MDN-0170 (1), was isolated from the culture broth of the marine-derived actinomycete strain CA-271078, together with three known related compounds identified as 4-dehydro-4a-dechloronapyradiomycin A1 (2), napyradiomycin A1 (3) and 3-chloro-6,8-dihydroxy-8-α-lapachone (4). The structure of the new compound was determined using a combination of spectroscopic techniques, including 1D and 2D NMR and electrospray-time of flight mass spectrometry (ESI-TOF MS). The relative configuration of compound 1, which contains two independent stereoclusters, has been established by molecular modelling in combination with nOe and coupling constant analyses. Biosynthetic arguments also allowed us to propose its absolute stereochemistry. The antimicrobial properties of the compounds isolated were evaluated against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Aspergillus fumigatus, and Candida albicans. The potent bioactivity previously reported for compounds 2 and 3 against methicillin-sensitive S. aureus has been extended to methicillin-resistant strains in this report. PMID:27763545

  19. Streptomyces aridus sp. nov., isolated from a high altitude Atacama Desert soil and emended description of Streptomyces melanogenes Sugawara and Onuma 1957

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A polyphasic study was undertaken to determine the taxonomic status of a Streptomyces strain which had been isolated from a high altitude Atacama Desert soil and shown to have bioactive properties. The strain, isolate H9T, was found to have chemotaxonomic, cultural, and morphological properties that...

  20. Antibacterial discovery in actinomycetes strains with mutations in RNA polymerase or ribosomal protein S12.

    PubMed

    Hosaka, Takeshi; Ohnishi-Kameyama, Mayumi; Muramatsu, Hideyuki; Murakami, Kana; Tsurumi, Yasuhisa; Kodani, Shinya; Yoshida, Mitsuru; Fujie, Akihiko; Ochi, Kozo

    2009-05-01

    We show that selection of drug-resistant bacterial mutants allows the discovery of antibacterial compounds. Mutant strains of a soil-isolated Streptomyces species that does not produce antibacterials synthesize a previously unknown class of antibacterial, which we name piperidamycin. Overall, 6% of non-Streptomyces actinomycetes species and 43% of Streptomyces species that do not produce antibacterials are activated to produce them. The antibacterial-producing mutants all carried mutations in RNA polymerase and/or the ribosomal protein S12.

  1. Genome Sequence of the Filamentous Actinomycete Kitasatospora viridifaciens

    PubMed Central

    Ramijan, Karina

    2017-01-01

    ABSTRACT The vast majority of antibiotics are produced by filamentous soil bacteria called actinomycetes. We report here the genome sequence of the tetracycline producer “Streptomyces viridifaciens” DSM 40239. Given that this species has the hallmark signatures characteristic of the Kitasatospora genus, we previously proposed to rename this organism Kitasatospora viridifaciens. PMID:28183757

  2. Efficacy of protease inhibitor from marine Streptomyces sp. VITBVK2 against Leishmania donovani - An in vitro study.

    PubMed

    Sreedharan, Veena; Bhaskara Rao, K V

    2017-03-01

    In the present study the leishmanicidal effect of potential protease inhibitor producing marine actinobacterial isolate has been investigated against Leishmania donovani, the causative agent of visceral leishmaniasis. Among 89 marine actinobacteria isolated from a salt pan in Kanyakumari, only one isolate (BVK2) showed 97% of protease inhibition activity against trypsin. Moderate to high protease inhibitor activity was shown by isolate BVK2 on proteinase (30%) and chymotrypsin (85%). In optimization study for protease inhibitor production glucose as carbon source and casein as nitrogen source showed the best activity. In the in-vitro Fluorescence-activated cell sorting (FACS) assay, 100 μg/ml of BVK2 extract was active against amastigotes in infected J774A.1 macrophages and showed 87% of parasitic inhibition. The isolate BVK2 showed significant anti-parasitic activity with an IC50 of 27.1 μg/ml after double doses were administered. The potential isolate was identified by molecular 16S rRNA gene sequencing as Streptomyces sp. VITBVK2. The results obtained suggest that the marine actinobacterial extract which have novel metabolites can be considered as a potential source for the development of drugs.

  3. Antibacterial and brine shrimp lethality effect of marine actinobacterium Streptomyces sp. CAS72 against human pathogenic bacteria

    PubMed Central

    Sivasankar, Palaniappan; Manivasagan, Panchanathan; Vijayanand, Packiyaraj; Sivakumar, Kannan; Sugesh, Shanmugam; Poongodi, Subramaniam; Maharani, Viswanathan; Vijayalakshmi, Shanmugam; Balasubramanian, Thangavel

    2013-01-01

    Objective To investigate the in vitro antibacterial activity against human pathogenic bacteria and brine shrimp lethality bioassay of the marine actinobacterium. Methods Forty six marine actinobacterial strains were isolated from sediment samples of Uppanar estuary, Cuddalore, India. Preliminary screening was done by cross-streak method and the potential strain was identified by morphological, chemotaxonomical and molecular methods. Fermentation was done and the metabolite was obtained by liquid-liquid extraction using ethyl acetate and purified by silica gel (100-200 mesh) column chromatography. The purified metabolite was tested for antibacterial activity, minimal inhibitory concentration and brine shrimp lethality bioassay. Results Among the forty six strains, CAS72 was found effective against human pathogenic bacteria. The strain CAS72 was identified as Streptomyces sp. The purified metabolite exhibited a significant in vitro antibacterial activity. The MIC value was also determined against human pathogenic bacteria and a strong cytotoxic activity in brine shrimp lethality assay was observed and the LC50 value was 23.5 µg/mL. Conclusions The present investigation reveals that the marine actinobacteria are well obtainable in Uppanar estuary environment and it can provide a definite source for novel bioactive metabolites.

  4. A xylanase from Streptomyces sp. FA1: heterologous expression, characterization, and its application in Chinese steamed bread.

    PubMed

    Xu, Yang; Wu, Jing; Zheng, Kaixuan; Wu, Dan

    2016-05-01

    Xylanases (EC 3.2.1.8) are hydrolytic enzymes that have found widespread application in the food, feed, and paper-pulp industries. Streptomyces sp. FA1 xynA was expressed as a secreted protein in Pichia pastoris, and the xylanase was applied to the production of Chinese steamed bread for the first time. The optimal pH and the optimal temperature of XynA were 5.5 and 60 °C, respectively. Using beechwood as substrate, the K m and V max were 2.408 mg mL(-1) and 299.3 µmol min(-1) mg(-1), respectively. Under optimal conditions, a 3.6-L bioreactor produced 1374 U mL(-1) of XynA activity at a protein concentration of 6.3 g L(-1) after 132 h of fermentation. Use of recombinant XynA led to a greater increase in the specific volume of the CSB than could be achieved using commercial xylanase under optimal conditions. This study provides the basis for the application of the enzyme in the baking industry.

  5. A New Analogue of Echinomycin and a New Cyclic Dipeptide from a Marine-Derived Streptomyces sp. LS298.

    PubMed

    Zhen, Xin; Gong, Ting; Liu, Fu; Zhang, Pei-Cheng; Zhou, Wan-Qi; Li, Yan; Zhu, Ping

    2015-11-18

    Quinomycin G (1), a new analogue of echinomycin, together with a new cyclic dipeptide, cyclo-(l-Pro-4-OH-l-Leu) (2), as well as three known antibiotic compounds tirandamycin A (3), tirandamycin B (4) and staurosporine (5), were isolated from Streptomyces sp. LS298 obtained from a marine sponge Gelliodes carnosa. The planar and absolute configurations of compounds 1 and 2 were established by MS, NMR spectral data analysis and Marfey's method. Furthermore, the differences in NMR data of keto-enol tautomers in tirandamycins were discussed for the first time. Antibacterial and anti-tumor activities of compound 1 were measured against 15 drug-sensitive/resistant strains and 12 tumor cell lines. Compound 1 exhibited moderate antibacterial activities against Staphylococcuse pidermidis, S. aureus, Enterococcus faecium, and E. faecalis with the minimum inhibitory concentration (MIC) values ranged from 16 to 64 μg/mL. Moreover, it displayed remarkable anti-tumor activities; the highest activity was observed against the Jurkat cell line (human T-cell leukemia) with an IC50 value of 0.414 μM.

  6. New action pattern of a maltose-forming alpha-amylase from Streptomyces sp. and its possible application in bakery.

    PubMed

    Ammar, Youssef Ben; Matsubara, Takayoshi; Ito, Kazuo; Iizuka, Masaru; Limpaseni, Tipaporn; Pongsawasdi, Piamsook; Minamiura, Noshi

    2002-11-30

    An a-Amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipitation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and 60 degrees C. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had alpha-anomeric forms, as determined by 1H-NMR. This maltose-forming alpha-Amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62% respectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.

  7. [Ecophysiological Characteristics of actinomycetes of desert soils of Mongolia].

    PubMed

    Zenova, G M; Kozhevin, P A; Manucharova, N A; Lubsanova, D A; Dubrova, M S

    2014-01-01

    It is shown that the actinomycete complex in steppe-desert light brown salty soil of desert steppes of Mongolia is represented by the genera Streptomyces and Micromonospora. The species diversity of the genus Streptomyces, which dominates the complex, decreases with increasing osmolarity of the medium. The influence of environmental factors--temperature and osmolarity of medium--on the development of metabolically active members of the phylum Actinobacteria in the domain Bacteria of the prokaryotic microbial soil community was established. The proportion of metabolically active bacteria belonging to Actinobacteria increases with increasing osmolarity and incubation temperature of soil. The dominance of the filamentous metabolically active members of the phylum Actinobacteria over the unicellular organisms was shown. The halotolerant actinomycetes isolated from the steppe-desert soils were alkalotolerant, xerophilic, and thermotolerant and exhibited antimicrobial activity with respect to Gram-positive bacteria and actinomycetes.

  8. Rhizocola hellebori gen. nov., sp. nov., an actinomycete of the family Micromonosporaceae containing 3,4-dihydroxydiaminopimelic acid in the cell-wall peptidoglycan.

    PubMed

    Matsumoto, Atsuko; Kawaguchi, Yoko; Nakashima, Takuji; Iwatsuki, Masato; Ōmura, Satoshi; Takahashi, Yōko

    2014-08-01

    An actinomycete strain, K12-0602(T), was isolated from the root of a Helleborus orientalis plant in Japan. The 16S rRNA gene sequence of strain K12-0602(T) showed that it had a close relationship with members of the family Micromonosporaceae and the 16S rRNA gene sequence similarity values between strain K12-0602(T) and type strains of type species of 27 genera belonging to the family Micromonosporaceae were below 96.2%. MK-9 (H4) and MK-9 (H6) were detected as major menaquinones, and galactose, xylose, mannose and ribose were present in the whole-cell hydrolysate. The acyl type of the peptidoglycan was glycolyl. Major fatty acids were iso-C(15 : 0), iso-C(16 : 0), C(17 : 1)ω9c and anteiso-C(17 : 0). Phosphatidylethanolamine was detected as the phospholipid corresponding to phospholipid type II. The G+C content of the genomic DNA was 67 mol%. Analyses of the cell-wall peptidoglycan by TLC and LC/MS showed that it was composed of alanine, glycine, hydroxylglutamic acid and an unknown amino acid, which was subsequently determined to be 3,4-dihydroxydiaminopimelic acid using instrumental analyses, including NMR and mass spectrometry. On the basis of the phylogenetic analysis and chemotaxonomic characteristics, strain K12-0602(T) represents a novel species of a new genus in the family Micromonosporaceae, for which the name Rhizocola hellebori gen. nov., sp. nov. is proposed. The type strain of the type species is K12-0602(T) ( = NBRC 109834(T) = DSM 45988(T)). This is the first report, to our knowledge, of 3,4-dihydroxydiaminopimelic acid being found as a diamino acid in bacterial cell-wall peptidoglycan.

  9. Extremophilic and extremotolerant actinomycetes in different soil types

    NASA Astrophysics Data System (ADS)

    Zenova, G. M.; Manucharova, N. A.; Zvyagintsev, D. G.

    2011-04-01

    Problems on the resistance of soil actinomycetes to various environmental factors (pH, salinity, temperature, and moisture) are discussed. Actinomycetes as a special group of prokaryotes were revealed to have a greater range of tolerance to these factors than was thought earlier. The regularities of the distribution of extremophilic and extremotolerant actinomycetes developing in unusual for mycelial bacteria conditions, their structural-functional characteristics, and their taxonomic composition were determined. The predominance of acidophilic representatives of the Micromonospora genus in acid soils (typical peat, soddy-podzolic, and taiga podzol) and the haloalkaliphilic Streptomyces pluricilirescens and S. prunicolor species in desert saline soils are shown. The specific features of the actinomycete complexes on thermal fields of the weakly developed stratified volcanic soils are described. In these complexes, the thermophilic forms were represented only by species of the Micromonospora genus; and the mesophilic forms, by Microbispora species. In the periodically heated desert soils, among the thermophilic actinomycetes, representatives of rare Actinomadura, Saccharopolyspora and Streptosporangium genera along with Streptomyces species were indicated. The mechanisms of the resistance of the actinomycetes to the extreme environmental conditions are discussed.

  10. The structural-functional organization of thermotolerant complexes of actinomycetes in desert and volcanic soils

    NASA Astrophysics Data System (ADS)

    Zenova, G. M.; Kurapova, A. I.; Lysenko, A. M.; Zvyagintsev, D. G.

    2009-05-01

    It has been found that the number of thermotolerant actinomycetes in strongly heated soils of deserts and volcanic regions is comparable to or exceeds the number of mesophilic actinomycetes. Among the latter group, streptomyces usually predominate; among thermotolerant actinomycetes, representatives of the Micromonospora, Streptosporangium, Actinomadura, Saccharopolyspora, Microtetraspora, and Microbispora genera are identified. Thermotolerant actinomycetes display the full cycle of their development in these soils. The method of fluorescent in situ hybridization has made it possible to determine that mycelial forms predominate among the metabolically active representatives of Actinobacteria; their portion increases with the rise in the temperature of soil incubation.

  11. An exo-β-(1→3)-D-galactanase from Streptomyces sp. provides insights into type II arabinogalactan structure

    PubMed Central

    Ling, Naomi X.-Y.; Lee, Joanne; Ellis, Miriam; Liao, Ming-Long; Mau, Shaio-Lim; Guest, David; Janssen, Peter H.; Kováč, Pavol; Bacic, Antony; Pettolino, Filomena A.

    2012-01-01

    An exo-β-(1→3)-D-galactanase (SGalase1) that specifically cleaves the β-(1→3)-D-galactan backbone of arabinogalactan-proteins (AGPs) was isolated from culture filtrates of a soil Streptomyces sp. Internal peptide sequence information was used to clone and recombinantly express the gene in E. coli. The molecular mass of the isolated enzyme was ~45 kDa, similar to the 48.2 kDa mass predicted from the amino acid sequence. The pI, pH and temperature optima for the enzyme were ~7.45, 3.8 and 48 °C, respectively. The native and recombinant enzymes specifically hydrolysed β-(1→3)-D-galacto-oligo- or poly-saccharides from the upstream (non-reducing) end, typical of an exo-acting enzyme. A second homologous Streptomyces gene (SGalase2) was also cloned and expressed. SGalase2 was similar in size (47.9 kDa) and enzyme activity to SGalase1 but differed in its pH optimum (pH 5). Both SGalase1 and SGalase2 are predicted to belong to the CAZy glycosyl hydrolase family GH 43 based on activity, sequence homology and phylogenetic analysis. The Km and Vmax of the native exo-β-(1→3)-D-galactanase for de-arabinosylated gum arabic (dGA) were 19 mg/ml and 9.7 μmol D-Gal/min/mg protein, respectively. The activity of these enzymes is well suited for the study of type II galactan structures and provides an important tool for the investigation of the biological role of AGPs in plants. De-arabinosylated gum arabic (dGA) was used as a model to investigate the use of these enzymes in defining type II galactan structure. Exhaustive hydrolysis of dGA resulted in a limited number of oligosaccharide products with a trisaccharide of Gal2GlcA1 predominating. PMID:22464224

  12. Strain improvement in actinomycetes in the postgenomic era.

    PubMed

    Baltz, Richard H

    2011-06-01

    With the recent advances in DNA sequencing technologies, it is now feasible to sequence multiple actinomycete genomes rapidly and inexpensively. An important observation that emerged from early Streptomyces genome sequencing projects was that each strain contains genes that encode 20 or more potential secondary metabolites, only a fraction of which are expressed during fermentation. More recently, this observation has been extended to many other actinomycetes with large genomes. The discovery of a wealth of orphan or cryptic secondary metabolite biosynthetic gene clusters has suggested that sequencing large numbers of actinomycete genomes may provide the starting materials for a productive new approach to discover novel secondary metabolites. The key issue for this approach to be successful is to find ways to turn on or turn up the expression of cryptic or poorly expressed pathways to provide material for structure elucidation and biological testing. In this review, I discuss several genetic approaches that are potentially applicable to many actinomycetes for this application.

  13. Purification and characterization of a novel lipopeptide from Streptomyces amritsarensis sp. nov. active against methicillin-resistant Staphylococcus aureus

    PubMed Central

    2014-01-01

    Nowadays antimicrobial lipopeptides are being widely exploited for developing potential therapeutic agents for treating bacterial infections. In the present study, we have purified and characterized an antimicrobial lipopeptide produced by Streptomyces amritsarensis sp. nov. (= MTCC 11845T = JCM 19660T). The lipopeptide was purified using silica gel chromatography, size exclusion chromatography and reverse phase- HPLC. The MS/MS analysis of the lipopeptide revealed that it has amino acid sequence as Ala-Thr-Gly-Ser-His-Gln and a long chain fatty acid tail with six times repeated the molecular mass of 161 Da which is corresponding to -C12H19. Based on the molecular mass (878.5 Da) and amino acid composition, the lipopeptide was identified as a novel lipopeptide. The MIC values of purified lipopeptide against Bacillus subtilis (MTCC 619), Staphylococcus epidermidis (MTCC 435), Mycobacterium smegmatis (MTCC 6) and clinical strain, Methicillin Resistant Staphylococcus aureus (MRSA) were found to be 10, 15, 25 and 45 μg/ml, respectively. It was completely stable at 70°C for 1 h and retained 81.8% activity after autoclaving (121°C for 15 min). It did not show any change in its activity profile between pH 5.0 - 9.0 and is stable to trypsin, proteinase K and lipase enzymes. It was found to be non-mutagenic against Salmonella typhimurium (TA98) and did not show cytotoxicity when checked against Chinese hamster ovary (CHO) cell line. In addition to antibacterial activity it also exhibits biosurfactant activity. PMID:25006539

  14. Paromomycin Derived from Streptomyces sp. AG-P 1441 Induces Resistance against Two Major Pathogens of Chili Pepper.

    PubMed

    Balaraju, Kotnala; Kim, Chang-Jin; Park, Dong-Jin; Nam, Ki-Woong; Zhang, Kecheng; Sang, Mee Kyung; Park, Kyungseok

    2016-09-28

    This is the first report that paromomycin, an antibiotic derived from Streptomyces sp. AG-P 1441 (AG-P 1441), controlled Phytophthora blight and soft rot diseases caused by Phytophthora capsici and Pectobacterium carotovorum, respectively, in chili pepper (Capsicum annum L.). Chili pepper plants treated with paromomycin by foliar spray or soil drenching 7 days prior to inoculation with P. capsici zoospores showed significant (p < 0.05) reduction in disease severity (%) when compared with untreated control plants. The disease severity of Phytophthora blight was recorded as 8% and 50% for foliar spray and soil drench, respectively, at 1.0 ppm of paromomycin, compared with untreated control, where disease severity was 83% and 100% by foliar spray and soil drench, respectively. A greater reduction of soft rot lesion areas per leaf disk was observed in treated plants using paromomycin (1.0 μg/ml) by infiltration or soil drench in comparison with untreated control plants. Paromomycin treatment did not negatively affect the growth of chili pepper. Furthermore, the treatment slightly promoted growth; this growth was supported by increased chlorophyll content in paromomycin-treated chili pepper plants. Additionally, paromomycin likely induced resistance as confirmed by the expression of pathogenesis-related (PR) genes: PR-1, β-1,3-glucanase, chitinase, PR-4, peroxidase, and PR-10, which enhanced plant defense against P. capsici in chili pepper. This finding indicates that AG-P 1441 plays a role in pathogen resistance upon the activation of defense genes, by secretion of the plant resistance elicitor, paromomycin.

  15. An unusual isopentenyl diphosphate isomerase found in the mevalonate pathway gene cluster from Streptomyces sp. strain CL190

    PubMed Central

    Kaneda, Kazuhide; Kuzuyama, Tomohisa; Takagi, Motoki; Hayakawa, Yoichi; Seto, Haruo

    2001-01-01

    A gene cluster encoding five enzymes of the mevalonate pathway had been cloned from Streptomyces sp. strain CL190. This gene cluster contained an additional ORF, orfD, encoding an unknown protein that was detected in some archaebacteria and some Gram-positive bacteria including Staphylococcus aureus. The recombinant product of orfD was purified as a soluble protein and characterized. The molecular mass of the enzyme was estimated to be 37 kDa by SDS-polyacrylamide gel electrophoresis and 155 kDa by gel filtration chromatography, suggesting that the enzyme is most likely to be a tetramer. The purified enzyme contained flavin mononucleotide (FMN) with the amount per tetramer being 1.4 to 1.6 mol/mol. The enzyme catalyzed the isomerization of isopentenyl diphosphate (IPP) to produce dimethylallyl diphosphate (DMAPP) in the presence of both FMN and NADPH. The Escherichia coli plasmid expressing orfD could complement the disrupted IPP isomerase gene in E. coli. These results indicate that orfD encodes an unusual IPP isomerase showing no sequence similarity to those of IPP isomerases identified to date. Based on the difference in enzymatic properties, we classify the IPP isomerases into two types: Type 2 for FMN- and NAD(P)H-dependent enzymes, and type 1 for the others. In view of the critical role of this isomerase in S. aureus and of the different enzymatic properties of mammalian (type 1) and S. aureus (type 2) isomerases, this unusual enzyme is considered to be a suitable molecular target for the screening of antibacterial drugs specific to S. aureus. PMID:11158573

  16. Partial Purification and Characterization of a Heat Stable α-Amylase from a Thermophilic Actinobacteria, Streptomyces sp. MSC702

    PubMed Central

    Singh, Renu; Kumar, Vijay; Kapoor, Vishal

    2014-01-01

    A partial purification and biochemical characterization of the α-amylase from Streptomyces sp. MSC702 were carried out in this study. The optimum operational conditions for enzyme substrate reaction for amylolytic enzyme activity from the strain were evaluated. The optimum pH, temperature, and incubation period for assaying the enzyme were observed to be 5.0, 55°C, and 30 min, respectively. The extracellular extract was concentrated using ammonium sulfate precipitation. It was stable in the presence of metal ions (5 mM) such as K+, Co2+, and Mo2+, whereas Pb2+, Mn2+, Mg2+, Cu2+, Zn2+, Ba2+, Ca2+, Hg2+, Sn2+, Cr3+, Al3+, Ag+, and Fe2+ were found to have inhibitory effects. The enzyme activity was also unstable in the presence of 1% Triton X-100, 1% Tween 80, 5 mM sodium lauryl sulphate, 1% glycerol, 5 mM EDTA, and 5 mM denaturant urea. At temperature 60°C and pH 5.0, the enzyme stability was maximum. α-amylase retained 100% and 34.18% stability for 1 h and 4 h, respectively, at 60°C (pH 7.0). The enzyme exhibited a half-life of 195 min at 60°C temperature. The analysis of kinetic showed that the enzyme has Km of 2.4 mg/mL and Vmax of 21853.0 μmol/min/mg for soluble potato starch. The results indicate that the enzyme reflects their potentiality towards industrial utilization. PMID:25400941

  17. Alloactinosynnema iranicum sp. nov., a rare actinomycete isolated from a hypersaline wetland, and emended description of the genus Alloactinosynnema.

    PubMed

    Nikou, Mahdi Moshtaghi; Ramezani, Mohaddaseh; Amoozegar, Mohammad Ali; Fazeli, Seyed Abolhassan Shahzadeh; Schumann, Peter; Spröer, Cathrin; Sánchez-Porro, Cristina; Ventosa, Antonio

    2014-04-01

    A Gram-staining-positive actinobacterial strain, Chem10(T), was isolated from soil around Inche-Broun hypersaline wetland in the north of Iran. Strain Chem10(T) was strictly aerobic, and catalase- and oxidase-positive. The isolate grew with 0-3 % NaCl, at 20-40 °C and at pH 6.0-8.0. The optimum temperature and pH for growth were 30 °C and pH 7.0, respectively. The cell wall of strain Chem10(T) contained meso-diaminopimelic acid as diamino acid and galactose, ribose and arabinose as whole-cell sugars. The polar lipid pattern contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain Chem10(T) synthesized cellular fatty acids of the straight-chain saturated and mono-unsaturated, and iso- and anteiso-branched types C14 : 0, C16 : 0, iso-C16 : 1, anteiso-C17 : 0, iso-C16 : 0, iso-C14 : 0 and iso-C15 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Chem10(T) belonged to the family Pseudonocardiaceae and showed the closest phylogenetic similarity to Alloactinosynnema album KCTC 19294(T) (98.3 %) and Actinokineospora cibodasensis DSM 45658(T) (97.9 %). DNA-DNA relatedness values between the novel strain and strains Alloactinosynnema album KCTC 19294(T) and Actinokineospora cibodasensis DSM 45658(T) were only 52 % and 23 %, respectively. On the basis of phylogenetic analysis, phenotypic characteristics and DNA-DNA hybridization data, a novel species of the genus Alloactinosynnema is proposed, Alloactinosynnema iranicum sp. nov. The type strain is Chem10(T) ( = IBRC-M 10403(T) = CECT 8209(T)). In addition, an emended description of the genus Alloactinosynnema is proposed.

  18. Presence of antioxidative agent, Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro- in newly isolated Streptomyces mangrovisoli sp. nov.

    PubMed Central

    Ser, Hooi-Leng; Palanisamy, Uma D.; Yin, Wai-Fong; Abd Malek, Sri N.; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2015-01-01

    A novel Streptomyces, strain MUSC 149T was isolated from mangrove soil. A polyphasic approach was used to study the taxonomy of MUSC 149T, which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the members of the genus Streptomyces. The diamino acid of the cell wall peptidoglycan was LL-diaminopimelic acid. The predominant menaquinones were identified as MK9(H8) and MK9(H6). Phylogenetic analysis indicated that closely related strains include Streptomyces rhizophilus NBRC 108885T (99.2% sequence similarity), S. gramineus NBRC 107863T (98.7%) and S. graminisoli NBRC 108883T (98.5%). The DNA–DNA relatedness values between MUSC 149T and closely related type strains ranged from 12.4 ± 3.3% to 27.3 ± 1.9%. The DNA G + C content was determined to be 72.7 mol%. The extract of MUSC 149T exhibited strong antioxidant activity and chemical analysis reported identification of an antioxidant agent, Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-. These data showed that metabolites of MUSC 149T shall be useful as preventive agent against free-radical associated diseases. Based on the polyphasic study of MUSC 149T, the strain merits assignment to a novel species, for which the name S. mangrovisoli sp. nov. is proposed. The type strain is MUSC 149T (=MCCC 1K00699T=DSM 100438T). PMID:26347733

  19. Cloning and characterization of the gene (farA) encoding the receptor for an extracellular regulatory factor (IM-2) from Streptomyces sp. strain FRI-5.

    PubMed Central

    Waki, M; Nihira, T; Yamada, Y

    1997-01-01

    IM-2 is a butyrolactone autoregulator that controls production of blue pigment and nucleoside antibiotics in Streptomyces sp. strain FRI-5. An IM-2-specific receptor gene, farA, was cloned from strain FRI-5, and nucleotide sequencing revealed that the farA gene consists of 666 bp encoding a 221-amino-acid protein of 24.3 kDa with an NH2-terminal amino acid sequence identical to that of purified native receptor. Another gene, farX, encoding a homolog of AfsA of Streptomyces griseus, was present upstream of farA. The monocistronic nature of the farA transcript was shown by Northern blot hybridization, and the transcript level increased upon addition of IM-2. Recombinant FarA expressed in and purified from E. coli showed clear ligand specificity toward IM-2, with a dissociation constant (Kd) for [3H]IM-2-C5 of 18.2 nM. FarA showed high overall homology to BarA (virginiae butanolide receptor from S. virginiae) and ArpA (A-factor receptor from S. griseus). Sequence alignment of the three receptor proteins revealed that the NH2-terminal region containing a helix-turn-helix DNA binding motif was highly conserved. The DNA binding motif is common in procaryotic repressors of the TetR family, suggesting that all the Streptomyces autoregulator receptors may act as transcriptional repressors. PMID:9260956

  20. Streptomyces malaysiense sp. nov.: A novel Malaysian mangrove soil actinobacterium with antioxidative activity and cytotoxic potential against human cancer cell lines.

    PubMed

    Ser, Hooi-Leng; Palanisamy, Uma Devi; Yin, Wai-Fong; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2016-04-13

    Actinobacteria from the unique intertidal ecosystem of the mangroves are known to produce novel, bioactive secondary metabolites. A novel strain known as MUSC 136(T) (=DSM 100712(T) = MCCC 1K01246(T)) which was isolated from Malaysian mangrove forest soil has proven to be no exception. Assessed by a polyphasic approach, its taxonomy showed a range of phylogenetic and chemotaxonomic properties consistent with the genus of Streptomyces. Phylogenetically, highest similarity was to Streptomyces misionensis NBRC 13063(T) (99.6%) along with two other strains (>98.9% sequence similarities). The DNA-DNA relatedness between MUSC 136(T) and these type strains ranged from 22.7 ± 0.5% to 46.5 ± 0.2%. Overall, polyphasic approach studies indicated this strain represents a novel species, for which the name Streptomyces malaysiense sp. nov. is proposed. The potential bioactivities of this strain were explored by means of antioxidant and cytotoxic assays. Intriguingly, MUSC 136(T) exhibited strong antioxidative activities as evaluated by a panel of antioxidant assays. It was also found to possess high cytotoxic effect against HCT-116 cells, which probably mediated through altering p53 protein and intracellular glutathione levels. Chemical analysis of the extract using GC-MS further affirms that the strain produces chemopreventive related metabolites.

  1. Streptomyces malaysiense sp. nov.: A novel Malaysian mangrove soil actinobacterium with antioxidative activity and cytotoxic potential against human cancer cell lines

    PubMed Central

    Ser, Hooi-Leng; Palanisamy, Uma Devi; Yin, Wai-Fong; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2016-01-01

    Actinobacteria from the unique intertidal ecosystem of the mangroves are known to produce novel, bioactive secondary metabolites. A novel strain known as MUSC 136T (=DSM 100712T = MCCC 1K01246T) which was isolated from Malaysian mangrove forest soil has proven to be no exception. Assessed by a polyphasic approach, its taxonomy showed a range of phylogenetic and chemotaxonomic properties consistent with the genus of Streptomyces. Phylogenetically, highest similarity was to Streptomyces misionensis NBRC 13063T (99.6%) along with two other strains (>98.9% sequence similarities). The DNA–DNA relatedness between MUSC 136T and these type strains ranged from 22.7 ± 0.5% to 46.5 ± 0.2%. Overall, polyphasic approach studies indicated this strain represents a novel species, for which the name Streptomyces malaysiense sp. nov. is proposed. The potential bioactivities of this strain were explored by means of antioxidant and cytotoxic assays. Intriguingly, MUSC 136T exhibited strong antioxidative activities as evaluated by a panel of antioxidant assays. It was also found to possess high cytotoxic effect against HCT-116 cells, which probably mediated through altering p53 protein and intracellular glutathione levels. Chemical analysis of the extract using GC-MS further affirms that the strain produces chemopreventive related metabolites. PMID:27072394

  2. Fatty acid biosynthesis in actinomycetes

    PubMed Central

    Gago, Gabriela; Diacovich, Lautaro; Arabolaza, Ana; Tsai, Shiou-Chuan; Gramajo, Hugo

    2011-01-01

    All organisms that produce fatty acids do so via a repeated cycle of reactions. In mammals and other animals, these reactions are catalyzed by a type I fatty acid synthase (FAS), a large multifunctional protein to which the growing chain is covalently attached. In contrast, most bacteria (and plants) contain a type II system in which each reaction is catalyzed by a discrete protein. The pathway of fatty acid biosynthesis in Escherichia coli is well established and has provided a foundation for elucidating the type II FAS pathways in other bacteria (White et al., 2005). However, fatty acid biosynthesis is more diverse in the phylum Actinobacteria: Mycobacterium, possess both FAS systems while Streptomyces species have only the multi-enzyme FAS II system and Corynebacterium species exclusively FAS I. In this review we present an overview of the genome organization, biochemical properties and physiological relevance of the two FAS systems in the three genera of actinomycetes mentioned above. We also address in detail the biochemical and structural properties of the acyl-CoA carboxylases (ACCases) that catalyzes the first committed step of fatty acid synthesis in actinomycetes, and discuss the molecular bases of their substrate specificity and the structure-based identification of new ACCase inhibitors with anti-mycobacterial properties. PMID:21204864

  3. Total synthesis of a reported fluorometabolite from Streptomyces sp. TC1 indicates an incorrect assignment. The isolated compound did not contain fluorine.

    PubMed

    Ayoup, Mohammed Salah; Cordes, David B; Slawin, Alexandra M Z; O'Hagan, David

    2014-06-27

    3,5-Di-tert-butyl-4-fluorophenylpropionic acid (1) was recently reported as a natural product from Streptomyces sp. TC1. This was a notable disclosure because fluorinated natural products are exceedingly rare, and in this case it suggested that the bacterium had the capacity to mediate an enzymatic aryl fluorination reaction. However, a synthesis of the putative metabolite 1 demonstrates that the spectroscopic data are inconsistent with the proposed structure. There is no evidence that the isolated compound contained a fluorine atom.

  4. Antifungal Streptomyces spp. Associated with the Infructescences of Protea spp. in South Africa

    PubMed Central

    Human, Zander R.; Moon, Kyuho; Bae, Munhyung; de Beer, Z. Wilhelm; Cha, Sangwon; Wingfield, Michael J.; Slippers, Bernard; Oh, Dong-Chan; Venter, Stephanus N.

    2016-01-01

    Common saprophytic fungi are seldom present in Protea infructescences, which is strange given the abundance of mainly dead plant tissue in this moist protected environment. We hypothesized that the absence of common saprophytic fungi in Protea infructescences could be due to a special symbiosis where the presence of microbes producing antifungal compounds protect the infructescence. Using a culture based survey, employing selective media and in vitro antifungal assays, we isolated antibiotic producing actinomycetes from infructescences of Protea repens and P. neriifolia from two geographically separated areas. Isolates were grouped into three different morphological groups and appeared to be common in the Protea spp. examined in this study. The three groups were supported in 16S rRNA and multi-locus gene trees and were identified as potentially novel Streptomyces spp. All of the groups had antifungal activity in vitro. Streptomyces sp. Group 1 had inhibitory activity against all tested fungi and the active compound produced by this species was identified as fungichromin. Streptomyces spp. Groups 2 and 3 had lower inhibition against all tested fungi, while Group 3 showed limited inhibition against Candida albicans and Sporothrix isolates. The active compound for Group 2 was also identified as fungichromin even though its production level was much lower than Group 1. The antifungal activity of Group 3 was linked to actiphenol. The observed antifungal activity of the isolated actinomycetes could contribute to protection of the plant material against common saprophytic fungi, as fungichromin was also detected in extracts of the infructescence. The results of this study suggest that the antifungal Streptomyces spp. could play an important role in defining the microbial population associated with Protea infructescences. PMID:27853450

  5. Moderately haloalkaliphilic actinomycetes in salt-affected soils

    NASA Astrophysics Data System (ADS)

    Zvyagintsev, D. G.; Zenova, G. M.; Oborotov, G. V.

    2009-12-01

    It was found that the population density of actinomycetes in solonchaks and saline desert soils varied from hundreds to tens of thousands of colony-forming units (CFUs) per 1 g of soil depending on soil type and was by 1-3 orders of magnitude lower than the number of mycelial bacteria in main soil types. Actinomycetes grow actively in saline soils, and the length of their mycelium reaches 140 m per 1 g of soil. Domination of moderately halophilic, alkaliphilic, and haloalkaliphilic actinomycetes, which grow well under 5% NaCl and pH 8-9, is a specific feature of actinomycetal complexes in saline soils. Representatives of Streptomyces and Micromonospora genera were found among the haloalkaliphilic actinomycetes. Micromonospores demonstrated lower (than streptomycetes) adaptability to high salt concentrations. Investigation of the phylogenetic position of isolated dominant haloalkaliphilic strains of streptomycetes performed on the basis of sequencing of the gene 16S rRNA enabled identifying these strains as Streptomyces pluricolorescens and S. prunicolor.

  6. Draft genome sequence of the marine Streptomyces sp. strain PP-C42, isolated from the Baltic Sea.

    PubMed

    Fan, Longjiang; Liu, Yun; Li, Zefeng; Baumann, Heike I; Kleinschmidt, Katrin; Ye, Wanzhi; Imhoff, Johannes F; Kleine, Michael; Cai, Daguang

    2011-07-01

    Streptomyces, a branch of aerobic Gram-positive bacteria, represents the largest genus of actinobacteria. The streptomycetes are characterized by a complex secondary metabolism and produce over two-thirds of the clinically used natural antibiotics today. Here we report the draft genome sequence of a Streptomyces strain, PP-C42, isolated from the marine environment. A subset of unique genes and gene clusters for diverse secondary metabolites as well as antimicrobial peptides could be identified from the genome, showing great promise as a source for novel bioactive compounds.

  7. Membrane-active metabolites produced by soil actinomycetes using chromatic phospholipid/polydiacetylene vesicles.

    PubMed

    Mehravar, Maryam; Sardari, Soroush; Owlia, Parviz

    2011-12-01

    Increased resistance of pathogens toward existing antibiotics has compelled the research efforts to introduce new antimicrobial substances. Drugs with new and less resistant-prone targets to antimicrobial activity have a high priority for drug development activities. Cell membrane seems to be a potential target for new antibiotic agent development to overcome resistance. In this study, A total number of 67 actinomycetes were isolated from the soil samples collected from desert, farming and mineral parts of Iran. We used a chromatic sensor as a membrane model that was set up for the target of antimicrobial metabolites of actinomycetes isolated from the soil. The sensors particles were composed of phospholipid and polymerized polydiacetylene (PDA) lipids. These polymers exhibited color change following interaction with membrane-active metabolites. The color change was due to structural disorder in the lipids following their interaction with membrane-active metabolites. The resultant color change was recorded by fluorescent microscope and easily recognizable by naked eye as well. Sixteen strains were isolated which produced antimicrobial metabolites and were effective against test microorganisms (Escherichia coli, Candida albicans and Saccharomyces cerevisiae ). A total number of 3 out of 16 strains produced membrane-active metabolites. These 3 strains were identified using 16s rRNA as Streptomyces sp and submitted to GenBank (accession no. JN180853; JN180854; JN180855).

  8. Actinomycetal complex of light sierozem on the Kopet-Dag piedmont plain

    NASA Astrophysics Data System (ADS)

    Zenova, G. M.; Zvyagintsev, D. G.; Manucharova, N. A.; Stepanova, O. A.; Chernov, I. Yu.

    2016-10-01

    The population density of actinomycetes in the samples of light sierozem from the Kopet Dag piedmont plain (75 km from Ashkhabad, Turkmenistan) reaches hundreds of thousand CFU/g soil. The actinomycetal complex is represented by two genera: Streptomyces and Micromonospora. Representatives of the Streptomyces genus predominate and comprise 73 to 87% of the actinomycetal complex. In one sample, representatives of the Micromonospora genus predominated in the complex (75%). The Streptomyces genus in the studied soil samples is represented by the species from several sections and series: the species of section Helvolo-Flavus series Helvolus represent the dominant component of the streptomycetal complex; their portion is up to 77% of all isolated actinomycetes. The species of other sections and series are much less abundant. Thus, the percentage of the Cinereus Achromogenes section in the actinomycetal complex does not exceed 28%; representatives of the Albus section Albus series, Roseus section Lavendulae-Roseus series, and Imperfectus section belong to rare species; they have been isolated not from all the studied samples of light sierozem, and their portion does not exceed 10% of the actinomycetal complex.

  9. Paper Mill Effluent Decolorization by Fifty Streptomyces Strains

    PubMed Central

    Hernández, Manuel; Rodríguez, Juana; Soliveri, Juan; Copa, José L.; Pérez, María I.; Arias, María E.

    1994-01-01

    Fifty actinomycete strains isolated from lignocellulosic substrates were examined for the ability to remove the color from a paper mill effluent obtained after semichemical alkaline pulping of wheat straw. Streptomyces sp. strains UAH 15, UAH 23, UAH 30, and UAH 51 were selected for their ability to decolorize the effluent in a liquid medium containing 1% (wt/vol) glycerol, 0.2% (wt/vol) ammonium sulfate, and 80% (vol/vol) effluent. The highest levels of decolorization achieved after the strains grew were 60 to 65%. Strains UAH 30 and UAH 51 were selected for further study because of their different patterns of effluent decolorization during growth. Fractionation of the decolorized effluent by gel permeation chromatography demonstrated that there were reductions in the levels of absorbance of the high- and medium-molecular-weight compounds. These fractions were mainly responsible for the color of the effluent, while the last fractions, the low-molecular-weight compounds, could have been responsible for the residual color of the decolorized effluent. Thin-layer chromatography revealed significant differences among the patterns of bands corresponding to the acidified supernatants obtained after precipitation of alkali-lignin from the effluent samples decolorized by different Streptomyces strains. Images PMID:16349426

  10. Actinomycete integrative and conjugative elements

    PubMed Central

    te Poele, Evelien M.; Bolhuis, Henk

    2008-01-01

    This paper reviews current knowledge on actinomycete integrative and conjugative elements (AICEs). The best characterised AICEs, pSAM2 of Streptomyces ambofaciens (10.9 kb), SLP1 (17.3 kb) of Streptomyces coelicolor and pMEA300 of Amycolatopsis methanolica (13.3 kb), are present as integrative elements in specific tRNA genes, and are capable of conjugative transfer. These AICEs have a highly conserved structural organisation, with functional modules for excision/integration, replication, conjugative transfer, and regulation. Recently, it has been shown that pMEA300 and the related elements pMEA100 of Amycolatopsis mediterranei and pSE211 of Saccharopolyspora erythraea form a novel group of AICEs, the pMEA-elements, based on the unique characteristics of their replication initiator protein RepAM. Evaluation of a large collection of Amycolatopsis isolates has allowed identification of multiple pMEA-like elements. Our data show that, as AICEs, they mainly coevolved with their natural host in an integrated form, rather than being dispersed via horizontal gene transfer. The pMEA-like elements could be separated into two distinct populations from different geographical origins. One group was most closely related to pMEA300 and was found in isolates from Australia and Asia and pMEA100-related sequences were present in European isolates. Genome sequence data have enormously contributed to the recent insight that AICEs are present in many actinomycete genera. The sequence data also provide more insight into their evolutionary relationships, revealing their modular composition and their likely combined descent from bacterial plasmids and bacteriophages. Evidence is accumulating that AICEs act as modulators of host genome diversity and are also involved in the acquisition of secondary metabolite clusters and foreign DNA via horizontal gene transfer. Although still speculative, these AICEs may play a role in the spread of antibiotic resistance factors into pathogenic bacteria

  11. Biosynthesis of Akaeolide and Lorneic Acids and Annotation of Type I Polyketide Synthase Gene Clusters in the Genome of Streptomyces sp. NPS554

    PubMed Central

    Zhou, Tao; Komaki, Hisayuki; Ichikawa, Natsuko; Hosoyama, Akira; Sato, Seizo; Igarashi, Yasuhiro

    2015-01-01

    The incorporation pattern of biosynthetic precursors into two structurally unique polyketides, akaeolide and lorneic acid A, was elucidated by feeding experiments with 13C-labeled precursors. In addition, the draft genome sequence of the producer, Streptomyces sp. NPS554, was performed and the biosynthetic gene clusters for these polyketides were identified. The putative gene clusters contain all the polyketide synthase (PKS) domains necessary for assembly of the carbon skeletons. Combined with the 13C-labeling results, gene function prediction enabled us to propose biosynthetic pathways involving unusual carbon-carbon bond formation reactions. Genome analysis also indicated the presence of at least ten orphan type I PKS gene clusters that might be responsible for the production of new polyketides. PMID:25603349

  12. Characterization of the Deep-Sea Streptomyces sp. SCSIO 02999 Derived VapC/VapB Toxin-Antitoxin System in Escherichia coli.

    PubMed

    Guo, Yunxue; Yao, Jianyun; Sun, Chenglong; Wen, Zhongling; Wang, Xiaoxue

    2016-07-01

    Toxin-antitoxin (TA) systems are small genetic elements that are ubiquitous in prokaryotes. Most studies on TA systems have focused on commensal and pathogenic bacteria; yet very few studies have focused on TAs in marine bacteria, especially those isolated from a deep sea environment. Here, we characterized a type II VapC/VapB TA system from the deep-sea derived Streptomyces sp. SCSIO 02999. The VapC (virulence-associated protein) protein belongs to the PIN (PilT N-terminal) superfamily. Overproduction of VapC strongly inhibited cell growth and resulted in a bleb-containing morphology in E. coli. The toxicity of VapC was neutralized through direct protein-protein interaction by a small protein antitoxin VapB encoded by a neighboring gene. Antitoxin VapB alone or the VapB/VapC complex negatively regulated the vapBC promoter activity. We further revealed that three conserved Asp residues in the PIN domain were essential for the toxic effect of VapC. Additionally, the VapC/VapB TA system stabilized plasmid in E. coli. Furthermore, VapC cross-activated transcription of several TA operons via a partially Lon-dependent mechanism in E. coli, and the activated toxins accumulated more preferentially than their antitoxin partners. Collectively, we identified and characterized a new deep sea TA system in the deep sea Streptomyces sp. and demonstrated that the VapC toxin in this system can cross-activate TA operons in E. coli.

  13. Characterization of the Deep-Sea Streptomyces sp. SCSIO 02999 Derived VapC/VapB Toxin-Antitoxin System in Escherichia coli

    PubMed Central

    Guo, Yunxue; Yao, Jianyun; Sun, Chenglong; Wen, Zhongling; Wang, Xiaoxue

    2016-01-01

    Toxin-antitoxin (TA) systems are small genetic elements that are ubiquitous in prokaryotes. Most studies on TA systems have focused on commensal and pathogenic bacteria; yet very few studies have focused on TAs in marine bacteria, especially those isolated from a deep sea environment. Here, we characterized a type II VapC/VapB TA system from the deep-sea derived Streptomyces sp. SCSIO 02999. The VapC (virulence-associated protein) protein belongs to the PIN (PilT N-terminal) superfamily. Overproduction of VapC strongly inhibited cell growth and resulted in a bleb-containing morphology in E. coli. The toxicity of VapC was neutralized through direct protein–protein interaction by a small protein antitoxin VapB encoded by a neighboring gene. Antitoxin VapB alone or the VapB/VapC complex negatively regulated the vapBC promoter activity. We further revealed that three conserved Asp residues in the PIN domain were essential for the toxic effect of VapC. Additionally, the VapC/VapB TA system stabilized plasmid in E. coli. Furthermore, VapC cross-activated transcription of several TA operons via a partially Lon-dependent mechanism in E. coli, and the activated toxins accumulated more preferentially than their antitoxin partners. Collectively, we identified and characterized a new deep sea TA system in the deep sea Streptomyces sp. and demonstrated that the VapC toxin in this system can cross-activate TA operons in E. coli. PMID:27376329

  14. Actinomycetes from the South China Sea sponges: isolation, diversity, and potential for aromatic polyketides discovery

    PubMed Central

    Sun, Wei; Zhang, Fengli; He, Liming; Karthik, Loganathan; Li, Zhiyong

    2015-01-01

    Marine sponges often harbor dense and diverse microbial communities including actinobacteria. To date no comprehensive investigation has been performed on the culturable diversity of the actinomycetes associated with South China Sea sponges. Structurally novel aromatic polyketides were recently discovered from marine sponge-derived Streptomyces and Saccharopolyspora strains, suggesting that sponge-associated actinomycetes can serve as a new source of aromatic polyketides. In this study, a total of 77 actinomycete strains were isolated from 15 South China Sea sponge species. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 12 families and 20 genera, among which three rare genera (Marihabitans, Polymorphospora, and Streptomonospora) were isolated from marine sponges for the first time. Subsequently, β-ketoacyl synthase (KSα) gene was used as marker for evaluating the potential of the actinomycete strains to produce aromatic polyketides. As a result, KSα gene was detected in 35 isolates related to seven genera (Kocuria, Micromonospora, Nocardia, Nocardiopsis, Saccharopolyspora, Salinispora, and Streptomyces). Finally, 10 strains were selected for small-scale fermentation, and one angucycline compound was detected from the culture extract of Streptomyces anulatus strain S71. This study advanced our knowledge of the sponge-associated actinomycetes regarding their diversity and potential in producing aromatic polyketides. PMID:26483773

  15. Soil actinomycetes in the National Forest Park in northeastern China

    NASA Astrophysics Data System (ADS)

    Shirokikh, I. G.; Shirokikh, A. A.

    2017-01-01

    The taxonomic and functional structure of actinomycete complexes in the litters and upper horizons of the soils under an artificial coniferous-broad-leaved forest located around the town of Chanchun (Tszilin province, PRC). The complex of actinomycetes included representatives of the Streptomyces, Micromonospora, Streptosporangium, and Streptoverticillium genera and oligosporous forms. In the actinomycete complexes, streptomycetes prevailed in the abundance (61-95%) and frequency of occurrence (100%). In the parcels of Korean pine ( Pinus koraiensis) and Mongolian oak ( Quercus mongolica), streptomycetes of 19 species from 8 series and 4 sections were isolated. The most representative, as in European forest biomes, was the Cinereus Achromogenes series. A distinguishing feature of the streptomycete complex in the biomes studied was the high participation of species from the Imperfectus series. The verification of the functional activity of natural isolates made it possible to reveal strains with high antagonistic and cellulolytic abilities. A high similarity of actinomycete complexes was found in Eurasian forest ecosystems remote from each other, probably due to the similarity of plant polymers decomposable by actinomycetes.

  16. Actinomycetes from Eucalyptus and their biological activities for controlling Eucalyptus leaf and shoot blight.

    PubMed

    Himaman, Winanda; Thamchaipenet, Arinthip; Pathom-Aree, Wasu; Duangmal, Kannika

    2016-01-01

    In Thailand, Eucalyptus plantations rapidly expand across the country. Leaf and shoot blight caused by Cryptosporiopsis eucalypti, Cylindrocladium sp. and Teratosphaeria destructans is a serious disease in Eucalyptus plantations. In this study, a total of 477 actinomycete strains were successfully isolated from roots and rhizosphere soil of Eucalyptus. Four hundred and thirty nine isolates were classified as streptomycetes and 38 isolates were non-streptomycetes. Among these isolates, 272 (57.0%), 118 (24.7%) and 241 (50.5%) isolates were antagonistic to Cryptosporiopsis eucalypti, Cylindrocladium sp. and Teratosphaeria destructans, respectively. All isolates were tested for their abilities to produce siderophores, indole acetic acid (IAA) and solubilise phosphate. Most isolates (464, 97.3%) produced siderophores. The majority of isolates (345, 72.3%) solubilised phosphate. In addition, almost half of these isolates (237, 49.7%) produced indole acetic acid. Strain EUSKR2S82 which showed the strongest inhibitory effect against all tested fungi with plant growth promoting ability was selected to test with Eucalyptus. This strain could colonize plant roots and increase Eucalyptus roots length. In a detached leaves bioassay, the disease severity of EUSKR2S82-inoculated Eucalyptus leaves was only 30% compared to 95% in the control treatment. The 16S rRNA gene sequence analysis revealed that the strain EUSKR2S82 was related to Streptomyces ramulosus NRRL-B 2714(T) (99.44% similarity). Identification of non-streptomycete isolates using 16S rRNA gene sequences classified them into 9 genera: Actinoallomurus, Actinomadura, Amycolatopsis, Cryptosporangium, Microbispora, Micromonospora, Nocardia, Nonomuraea and Pseudonocardia. It is evident that Eucalyptus tree harbored several genera of actinomycetes. The selected isolate, EUSKR2S82 showed potential as a candidate for biocontrol agent of leaf and shoot blight of Eucalyptus and to promote growth.

  17. Evaluation of Streptomyces sp. strain g10 for suppression of Fusarium wilt and rhizosphere colonization in pot-grown banana plantlets.

    PubMed

    Getha, K; Vikineswary, S; Wong, W H; Seki, T; Ward, A; Goodfellow, M

    2005-01-01

    Streptomyces sp. strain g10 exhibited strong antagonism towards Fusarium oxysporum f.sp. cubense (Foc) races 1, 2 and 4 in plate assays by producing extracellular antifungal metabolites. Treating the planting hole and roots of 4-week-old tissue-culture-derived 'Novaria' banana plantlets with strain g10 suspension (10(8) cfu/ml), significantly (P < 0.05) reduced wilt severity when the plantlets were inoculated with 10(4) spores/ml Foc race 4. The final disease severity index for leaf symptom (LSI) and rhizome discoloration (RDI) was reduced about 47 and 53%, respectively, in strain g10-treated plantlets compared to untreated plantlets. Reduction in disease incidence was not significant (P < 0.05) when plantlets were inoculated with a higher concentration (10(6) spores/ml) of Foc race 4. Rhizosphere population of strain g10 showed significant (P = 0.05) increase of more than 2-fold at the end of the 3rd week compared to the 2nd week after soil amendment with the antagonist. Although the level dropped, the rhizosphere population at the end of the 6th week was still nearly 2-fold higher than the level detected after 2 weeks. In contrast, the root-free population declined significantly (P = 0.05), nearly 4-fold after 6 weeks when compared to the level detected after 2 weeks. Neither growth-inhibiting nor growth-stimulating effects were observed in plantlets grown in strain g10-amended soil.

  18. Harnessing the Potential of Halogenated Natural Product Biosynthesis by Mangrove-Derived Actinomycetes

    PubMed Central

    Li, Xue-Gong; Tang, Xiao-Min; Xiao, Jing; Ma, Guang-Hui; Xu, Li; Xie, Shu-Jie; Xu, Min-Juan; Xiao, Xiang; Xu, Jun

    2013-01-01

    Mangrove-derived actinomycetes are promising sources of bioactive natural products. In this study, using homologous screening of the biosynthetic genes and anti-microorganism/tumor assaying, 163 strains of actinomycetes isolated from mangrove sediments were investigated for their potential to produce halogenated metabolites. The FADH2-dependent halogenase genes, identified in PCR-screening, were clustered in distinct clades in the phylogenetic analysis. The coexistence of either polyketide synthase (PKS) or nonribosomal peptide synthetase (NRPS) as the backbone synthetases in the strains harboring the halogenase indicated that these strains had the potential to produce structurally diversified antibiotics. As a validation, a new enduracidin producer, Streptomyces atrovirens MGR140, was identified and confirmed by gene disruption and HPLC analysis. Moreover, a putative ansamycin biosynthesis gene cluster was detected in Streptomyces albogriseolus MGR072. Our results highlight that combined genome mining is an efficient technique to tap promising sources of halogenated natural products synthesized by mangrove-derived actinomycetes. PMID:24129229

  19. Harnessing the potential of halogenated natural product biosynthesis by mangrove-derived actinomycetes.

    PubMed

    Li, Xue-Gong; Tang, Xiao-Min; Xiao, Jing; Ma, Guang-Hui; Xu, Li; Xie, Shu-Jie; Xu, Min-Juan; Xiao, Xiang; Xu, Jun

    2013-10-14

    Mangrove-derived actinomycetes are promising sources of bioactive natural products. In this study, using homologous screening of the biosynthetic genes and anti-microorganism/tumor assaying, 163 strains of actinomycetes isolated from mangrove sediments were investigated for their potential to produce halogenated metabolites. The FADH2-dependent halogenase genes, identified in PCR-screening, were clustered in distinct clades in the phylogenetic analysis. The coexistence of either polyketide synthase (PKS) or nonribosomal peptide synthetase (NRPS) as the backbone synthetases in the strains harboring the halogenase indicated that these strains had the potential to produce structurally diversified antibiotics. As a validation, a new enduracidin producer, Streptomyces atrovirens MGR140, was identified and confirmed by gene disruption and HPLC analysis. Moreover, a putative ansamycin biosynthesis gene cluster was detected in Streptomyces albogriseolus MGR072. Our results highlight that combined genome mining is an efficient technique to tap promising sources of halogenated natural products synthesized by mangrove-derived actinomycetes.

  20. Larvicidal potential of Asteraceae family endophytic actinomycetes against Culex quinquefasciatus mosquito larvae.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2014-01-01

    Pakistan is blessed with plants of Asteraceae family with known medicinal background used for centuries by Hakims (traditional physicians). Keeping in mind the background of their anti-larval potential, a total of 21 endophytic actinomycetes were isolated from four Asteraceae plants and screened against the first and fourth instar stages of Culex quinquefasciatus Say mosquito larvae. Of the 21 isolates, 6 of them gave strong larvicidal activity (80-100% mortality) in the screening results and 4 isolates gave a potent larvicidal activity (100% mortality) at the fourth instar stage. These isolates belonged to different species within the actinomycetes group, namely Streptomyces albovinaceus and Streptomyces badius. This communication reports the larvicidal potential of endophytic actinomycetes residing within the native Asteraceae plants in Pakistan. The study suggests further exploration through large-scale productions leading to the identification of the larvicidal compounds.

  1. Isolation and characterization of medically important aerobic actinomycetes in soil of iran (2006 - 2007).

    PubMed

    Aghamirian, Mohammad Reza; Ghiasian, Seyed Amir

    2009-01-01

    The aerobic actinomycetes are a large group of soil-inhabiting bacteria that occur worldwide. Some of them are the main cause of two important diseases, nocardiosis and actinomycetoma. To identify the prevalence and geographic distribution of aerobic actinomycetes in soil of Qazvin province, a study was carried out during 2006-2007. In this study, the incidence and diversity of medically important aerobic actinomycetes was determined in 300 soil samples of different parts of Qazvin. The suspensions of superficial soil samples were prepared by adding of normal saline, streptomycin and chloramphenicol and the supernatants were cultured on brain-heart infusion agar and Sabouraud's dextrose agar contain cycloheximide. The isolated microorganisms were examined by Gram and acid-fast stains and were identified biochemically and morphologically. Of 96 aerobic actinomycetes isolates identified, Actinomadura madurae and Streptomyces somaliensis were the most frequently isolated species each representing 19.8% of isolates, followed by Nocardia asteroides (15.6%), N. otitidiscaviarum (9.4%), N. brasiliensis (7.3%), A. peletieri, S. griseus, and Nocardia spp. (each 5.2%), and N. transvalensis, Nocardiopsis dassonvillei, Actinomadura spp. and Streptomyces spp. (each 3.1%). To the best of our knowledge, this is the first report on epidemiological investigation of medically important aerobic actinomycetes in soil samples from Iran. In recent years, mycetoma and nocardiosis have been increasingly reported in Iran. The results showed that medically important actinomycetes occur in the environment of Iran and soil could be potential source of actinomycotic infections.

  2. Bioactive Potential of Actinomycetes from Less Explored Ecosystems against Mycobacterium tuberculosis and Other Nonmycobacterial Pathogens

    PubMed Central

    Venugopal, Gopikrishnan; Subramaniam, Balaji; Ramasamy, Balagurunathan

    2014-01-01

    Bioactive potential of actinomycetes isolated from certain less explored Indian ecosystems against Mycobacterium tuberculosis and other nonmycobacterial pathogens was investigated. Actinomycetes were isolated from the soil samples collected from desert, coffee plantation, rubber forest, and hill area and their cultural and micromorphological characteristics were studied. Crude extracts were prepared by agar surface fermentation and tested against M. tuberculosis isolates by luciferase reporter phage (LRP) assay at 100 µg/mL. Activity against nonmycobacterial pathogens was studied by agar plug method. Totally 54 purified cultures of actinomycetes including 43 Streptomyces and 11 non-Streptomyces were isolated. While screening for antitubercular activity, extracts of 39 actinomycetes showed activity against one or more M. tuberculosis isolates whereas 27 isolates exhibited antagonistic activity against nonmycobacterial pathogens. In particular crude extracts from sixteen actinomycete isolates inhibited all the three M. tuberculosis isolates tested. Findings of the present study concluded that less explored ecosystems investigated in this study are the potential resource for bioactive actinomycetes. Further purification and characterization of active molecule from the potential extracts will pave the way for determination of MIC, toxicity, and specificity studies. PMID:27437460

  3. Diversity, bioactivities, and metabolic potentials of endophytic actinomycetes isolated from traditional medicinal plants in Sichuan, China.

    PubMed

    Qiu, Peng; Feng, Zhi-Xiang; Tian, Jie-Wei; Lei, Zu-Chao; Wang, Lei; Zeng, Zhi-Gang; Chu, Yi-Wen; Tian, Yong-Qiang

    2015-12-01

    The present study was designed to determine the taxonomic diversity and metabolic activity of the actinomycetes community, including 13 traditional medicinal plants collected in Sichuan province, China, using multiple approaches such as morphological and molecular identification methods, bioactivity assays, and PCR screening for genes involved in antibiotics biosynthesis. 119 endophytic actinomycetes were recovered; 80 representative strains were chosen for 16S rRNA gene partial sequence analyses, with 66 of them being affiliated to genus Streptomyces and the remaining 14 strains being rare actinomycetes. Antimicrobial tests showed that 12 (15%) of the 80 endophytic actinomycetes displayed inhibitory effects against at least one indicator pathogens, which were all assigned to the genus Streptomyces. In addition, 87.5% and 58.8% of the isolates showed anticancer and anti-diabetic activities, respectively. Meanwhile, the anticancer activities of the isolates negatively correlated with their anti-diabetic activities. Based on the results of PCR screening, five genes, PKS-I, PKS-II, NRPS, ANSA, and oxyB, were detected in 55.0%, 58.8%, 90.0%, 18.8% and 8.8% of the 80 actinomycetes, respectively. In conclusion, the PCR screening method employed in the present study was conducive for screening and selection of potential actinomycetes and predicting potential secondary metabolites, which could overcome the limitations of traditional activity screening models.

  4. Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. glycine.

    PubMed

    Mingma, Ratchanee; Pathom-aree, Wasu; Trakulnaleamsai, Savitr; Thamchaipenet, Arinthip; Duangmal, Kannika

    2014-01-01

    In this study, actinomycetes from roots and rhizospheric soils of leguminous plants were isolated using starch casein agar supplemented with antifungal and antibacterial antibiotics. Three hundred and seventeen actinomycetes were isolated with 77 isolates obtained from plant roots and 240 isolates from rhizospheric soils. Analysis of whole-organism hydrolysates showed that 289 strains were rich in the LL-isomer of diaminopimelic acid, a result consistent with their assignment to the streptomycetes. The remaining 28 strains were assigned to non-streptomycetes based on the presence of meso-isomer of diaminopimelic acid in cell wall. Sixty-four isolates (20.2%) showed antagonistic activity against soybean pathogen Xanthomonas campestris pv. glycine by agar overlay method. Isolate RM 365 showed the highest activity with an inhibition ratio of 3.79, with no inhibitory activity on the growth of Rhizobium japonicum TISTR 079, Rhizobium sp. TISTR 061 and Rhizobium sp. TISTR 063. The 16S rRNA gene sequence analysis revealed that isolate RM 365 shared 99.28% similarity to Streptomyces caeruleatus GIMN4(T) (GQ329712). In addition, isolates which contained meso-DAP were also identified by 16S rRNA gene sequence analysis. The results showed that they were members of the genus Amycolatopsis, Isoptericola, Micromonospora, Microbispora, Nocardia, Nonomuraea, Promicromonospora and Pseudonocardia.

  5. Actinomycetes inhibit filamentous fungi from the cuticle of Acromyrmex leafcutter ants.

    PubMed

    Dângelo, Rômulo Augusto Cotta; de Souza, Danival José; Mendes, Thais Demarchi; Couceiro, Joel da Cruz; Lucia, Terezinha Maria Castro Della

    2016-03-01

    Actinomycetes bacteria associated with leafcutter ants produce secondary metabolites with antimicrobial properties against Escovopsis, a fungus specialized in attacking the gardens of fungus-growing ants, which denies the ants their food source. Because previous studies have used fungi isolated from fungus gardens but not from ant integument, the aims of the present study were to isolate actinomycetes associated with the cuticle of the Acromyrmex spp. and to quantify their inhibition abilities against the filamentous fungal species carried by these ants. The results demonstrated that actinomycetes had varied strain-dependent effects on several filamentous fungal species in addition to antagonistic activity against Escovopsis. The strain isolated from Acromyrmex balzani was identified as a Streptomyces species, whereas the remaining isolates were identified as different strains belonging to the genus Pseudonocardia. These findings corroborate the hypothesis that actinomycetes do not act specifically against Escovopsis mycoparasites and may have the ability to inhibit other species of pathogenic fungi.

  6. Diversity of actinomycetes isolated from subseafloor sediments after prolonged low-temperature storage.

    PubMed

    Ulanova, Dana; Goo, Kian-Sim

    2015-05-01

    Subseafloor sediments present an untapped source of novel bacterial species with industrially important bioactivities. Subseafloor core samples collected during the Integrated Ocean Drilling Program Expeditions 315, 316, and 331 and stored in Kochi Core Center at -80 °C for 1 to 4 years were used for cultivation-based study of viable actinomycetes. In total, more than 100 actinomycete-like colonies were isolated from two deep-frozen subseafloor sediment samples. Isolated actinomycetes showed close similarity to known Actinotalea, Dietzia, Gordonia, Isoptericola, Microbacterium, Nocardia, Rhodococcus, Pseudonocardia, Streptomyces, and Tsukamurella species and were halotolerant. Bioactivity assays revealed that two of the isolates were producing potent antibacterial compound(s) and one isolate was having antifungal activity. Our study demonstrated that deep-frozen subseafloor core samples could be a potential source of viable actinomycetes, which may be used in drug discovery.

  7. Fusion of Dioxygenase and Lignin-binding Domains in a Novel Secreted Enzyme from Cellulolytic Streptomyces sp. SirexAA-E*

    PubMed Central

    Bianchetti, Christopher M.; Harmann, Connor H.; Takasuka, Taichi E.; Hura, Gregory L.; Dyer, Kevin; Fox, Brian G.

    2013-01-01

    Streptomyces sp. SirexAA-E is a highly cellulolytic bacterium isolated from an insect/microbe symbiotic community. When grown on lignin-containing biomass, it secretes SACTE_2871, an aromatic ring dioxygenase domain fused to a family 5/12 carbohydrate-binding module (CBM 5/12). Here we present structural and catalytic studies of this novel fusion enzyme, thus providing insight into its function. The dioxygenase domain has the core β-sandwich fold typical of this enzyme family but lacks a dimerization domain observed in other intradiol dioxygenases. Consequently, the x-ray structure shows that the enzyme is monomeric and the Fe(III)-containing active site is exposed to solvent in a shallow depression on a planar surface. Purified SACTE_2871 catalyzes the O2-dependent intradiol cleavage of catechyl compounds from lignin biosynthetic pathways, but not their methylated derivatives. Binding studies show that SACTE_2871 binds synthetic lignin polymers and chitin through the interactions of the CBM 5/12 domain, representing a new binding specificity for this fold-family. Based on its unique structural features and functional properties, we propose that SACTE_2871 contributes to the invasive nature of the insect/microbial community by destroying precursors needed by the plant for de novo lignin biosynthesis as part of its natural wounding response. PMID:23653358

  8. New antimicrobial anthraquinone 6,6(1)-bis (1,5,7-trihydroxy-3-hydroxymethylanthraquinone) isolated from Streptomyces sp. isolate ERI-26.

    PubMed

    Duraipandiyan, V; Al-Dhabi, Naif Abdullah; Ignacimuthu, S

    2016-11-01

    The present report is about Streptomyces sp. isolate ERI-26 isolated from the soil sample of Nilgiri forest, Western Ghats. The methanol extract of ERI-26 showed good antimicrobial activity against tested microbes. The antimicrobial novel anthraquinones were purified by bioactivity-guided fractionation using a silica gel column and preparative HPLC. The compound was characterized and identified by UV, IR, NMR and MASS spectral data. The compound named as 6,6(1)-bis (1,5,7-trihydroxy-3-hydroxymethylanthraquinone), showed significant antimicrobial activities against tested microbes. The isolated compound inhibited the tested bacterial growth, Staphylococcus aureus at 62.5 μg/ml, Staphylococcus epidermidis at 15.62 μg/m, Bacillus subtilis at 62.5 μg/ml, fungi; Trichophyton mentagrophytes at 15.62 μg/m Trichophyton simii at 15.62 μg/ml, Aspergillus niger at. 7.81 μg/ml, Aspergiller flavus at 3.90 μg/ml, Trichophyton rubrum 296 at 62.5 μg/ml, T. rubrum 57/01 at 7.81 μg/ml, Magnaporthe grisea at 15.62 μg/ml. and Botrytis cinerea at 3.90 μg/ml. Isolated anthraquinone compound and its antimicrobial activity were newly reported.

  9. Biochemical Properties and Atomic Resolution Structure of a Proteolytically Processed β-Mannanase from Cellulolytic Streptomyces sp. SirexAA-E

    PubMed Central

    Takasuka, Taichi E.; Acheson, Justin F.; Bianchetti, Christopher M.; Prom, Ben M.; Bergeman, Lai F.; Book, Adam J.; Currie, Cameron R.; Fox, Brian G.

    2014-01-01

    β-mannanase SACTE_2347 from cellulolytic Streptomyces sp. SirexAA-E is abundantly secreted into the culture medium during growth on cellulosic materials. The enzyme is composed of domains from the glycoside hydrolase family 5 (GH5), fibronectin type-III (Fn3), and carbohydrate binding module family 2 (CBM2). After secretion, the enzyme is proteolyzed into three different, catalytically active variants with masses of 53, 42 and 34 kDa corresponding to the intact protein, loss of the CBM2 domain, or loss of both the Fn3 and CBM2 domains. The three variants had identical N-termini starting with Ala51, and the positions of specific proteolytic reactions in the linker sequences separating the three domains were identified. To conduct biochemical and structural characterizations, the natural proteolytic variants were reproduced by cloning and heterologously expressed in Escherichia coli. Each SACTE_2347 variant hydrolyzed only β-1,4 mannosidic linkages, and also reacted with pure mannans containing partial galactosyl- and/or glucosyl substitutions. Examination of the X-ray crystal structure of the GH5 domain of SACTE_2347 suggests that two loops adjacent to the active site channel, which have differences in position and length relative to other closely related mannanases, play a role in producing the observed substrate selectivity. PMID:24710170

  10. Characteristics of α-L-arabinofuranosidase from Streptomyces sp I10-1 for production of L-arabinose from corn hull arabinoxylan.

    PubMed

    Kurakake, Masahiro; Kanbara, Yoshikazu; Murakami, Yoshiki

    2014-03-01

    Streptomyces sp I10-1 α-L-arabinofuranosidase efficiently produced L-arabinose from high arabinose-content corn hull arabinoxylan (ratio of arabinose to xylose, 0.6). The optimum pH at 40 °C was around 6, and the enzyme was stable from pH 5 to 11. The optimum temperature was 50 °C at pH 5, and the activity was stable at 40 °C. The enzymatic activity against corn hull arabinoxylan was 2.3 times higher than towards p-nitrophenyl-α-L-arabinofuranoside. Approximately 45% L-arabinose recovery was achieved from corn hull arabinoxylan. It was considered that L-arabinose residues not removed by the enzyme were attributable to those linked with ferulic acid. The open reading frame of the enzyme gene consisted of 1,224 bp, and the predicted peptide was 408 amino acids, which corresponded to a molecular size of 45, 248 Da. It was presumed that the smaller molecular size (31,000 Da) estimated on SDS-PAGE resulted from proteolysis by proteases. I10-1 α-L-arabinofuranosidase belongs to the Alpha-L-AF C superfamily, which is associated with glycoside hydrolase family 51, but the properties were unique.

  11. Interaction with mycorrhiza helper bacterium Streptomyces sp. AcH 505 modifies organisation of actin cytoskeleton in the ectomycorrhizal fungus Amanita muscaria (fly agaric).

    PubMed

    Schrey, Silvia D; Salo, Vanamo; Raudaskoski, Marjatta; Hampp, Rüdiger; Nehls, Uwe; Tarkka, Mika T

    2007-08-01

    The actin cytoskeleton (AC) of fungal hyphae is a major determinant of hyphal shape and morphogenesis, implicated in controlling tip structure and secretory vesicle delivery. Hyphal growth of the ectomycorrhizal fungus Amanita muscaria and symbiosis formation with spruce are promoted by the mycorrhiza helper bacterium Streptomyces sp. AcH 505 (AcH 505). To investigate structural requirements of growth promotion, the effect of AcH 505 on A. muscaria hyphal morphology, AC and actin gene expression were studied. Hyphal diameter and mycelial density decreased during dual culture (DC), and indirect immunofluorescence microscopy revealed that the dense and polarised actin cap in hyphal tips of axenic A. muscaria changes to a loosened and dispersed structure in DC. Supplementation of growth medium with cell-free bacterial supernatant confirmed that reduction in hyphal diameter and AC changes occurred at the same stage of growth. Transcript levels of both actin genes isolated from A. muscaria remained unaltered, indicating that AC changes are regulated by reorganisation of the existing actin pool. In conclusion, the AC reorganisation appears to result in altered hyphal morphology and faster apical extension. The thus improved spreading of hyphae and increased probability to encounter plant roots highlights a mechanism behind the mycorrhiza helper effect.

  12. Purification and characterization of a thermostable keratinolytic serine alkaline proteinase from Streptomyces sp. strain AB1 with high stability in organic solvents.

    PubMed

    Jaouadi, Bassem; Abdelmalek, Badis; Fodil, Djamila; Ferradji, Fatma Zohra; Rekik, Hatem; Zaraî, Nedia; Bejar, Samir

    2010-11-01

    A keratinolytic alkaline proteinase (KERAB) was isolated from Streptomyces sp. strain AB1. Based on MALDI-TOF mass spectrometry analysis, the purified enzyme is a monomer with a molecular mass of 29850.17Da. The NH(2)-terminal sequence of the enzyme was determined to be TQANPPSWGLDDIDQTAL. This keratinase was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DIFP), which suggests that it belongs to the serine protease family. Using keratin azure as a substrate, the optimum pH and temperature values for keratinase activity were pH 11.5 and 75 degrees C, respectively. This keratinase was stable between 30 and 60 degrees C and pH 4 and 11 for 4 and 96 h, respectively, and thermoactivity and thermostability were enhanced in the presence of 5 mM Mg(2+). Its catalytic efficiency was higher than those of SAPB-L31I/T33S/N99Y, nattokinase and subtilisin Carlsberg. KERAB exhibited stability to detergents and high resistance against organic solvents and was able to degrade feathers completely. These properties make KERAB a potential candidate for future applications in detergent formulations, dehairing during leather processing, and non-aqueous peptide biocatalysis.

  13. Production of a Novel Fucoidanase for the Green Synthesis of Gold Nanoparticles by Streptomyces sp. and Its Cytotoxic Effect on HeLa Cells

    PubMed Central

    Manivasagan, Panchanathan; Oh, Junghwan

    2015-01-01

    Marine actinobacteria-produced fucoidanases have received considerable attention as one of the major research topics in recent years, particularly for the medical exploitation of fucoidans and their degradation products. The present study describes the optimization and production of a novel fucoidanase for the green synthesis of gold nanoparticles and its biological applications. The production of fucoidanase was optimized using Streptomyces sp. The medium components were selected in accordance with the Plackett-Burman design and were further optimized via response surface methodology. The fucoidanase was statistically optimized with the most significant factors, namely wheat bran 3.3441 g/L, kelp powder 0.7041 g/L, and NaCl 0.8807 g/L, respectively. The biosynthesized gold nanoparticles were determined by UV-vis spectroscopy and were further characterized by X-ray diffraction analysis, Fourier transform infrared spectroscopy, field emission scanning electron microscopy, energy dispersive X-ray analysis, and high-resolution transmission electron microscopy. Furthermore, the biosynthesized gold nanoparticles exhibited a dose-dependent cytotoxicity against HeLa cells and the inhibitory concentration (IC50) was found to be 350 µg/mL at 24 h and 250 µg/mL at 48 h. Therefore, the production of novel fucoidanase for the green synthesis of gold nanoparticles has comparatively rapid, less expensive and wide application to anticancer therapy in modern medicine. PMID:26569267

  14. In silico molecular docking, preclinical evaluation of spiroindimicins A-D, lynamicin A and D isolated from deep marine sea derived Streptomyces sp. SCSIO 03032.

    PubMed

    Saurav, Kumar; Zhang, Wenjun; Saha, Subhasish; Zhang, Haibo; Li, Sumei; Zhang, Qingbo; Wu, Zhengchao; Zhang, Guangtao; Zhu, Yiguang; Verma, Gaurav

    2014-09-01

    The criteria used for successful drug discovery involves high throughput screening for preclinical evaluation and its interaction with target enzymes. In silico approach resulting in the creation of drug like library and identification of essential reactions and pathways spreads across several parts of metabolism. The aim of the present study was to evaluate the preclinical property and interaction to various drug target enzymes for spiroindimicins A-D and lynamicin A and D isolated from deep marine sea derived Streptomyces sp. SCSIO 03032 with 7 selected drug target enzymes. The preclinical and molecular docking simulation was performed using In silico pharmacology and docking tool. Drug likeliness, ADME and toxicity testing findings suggested the compounds with oral drug candidate's probability. Interaction of isolated compounds against drug target enzymes was satisfactory with Spiroindimicins C, D and Lynamicin D emerging as most potent Topoisomerase II, Cathepsin K, Cytochrome P4503A4, Aromatase P450, protein kinase and histone deacetylase inhibitors. Our results suggest that In silico approach in drug discovery procedure in later stage of development can ease up making lead molecules library.

  15. Production of a Novel Fucoidanase for the Green Synthesis of Gold Nanoparticles by Streptomyces sp. and Its Cytotoxic Effect on HeLa Cells.

    PubMed

    Manivasagan, Panchanathan; Oh, Junghwan

    2015-11-12

    Marine actinobacteria-produced fucoidanases have received considerable attention as one of the major research topics in recent years, particularly for the medical exploitation of fucoidans and their degradation products. The present study describes the optimization and production of a novel fucoidanase for the green synthesis of gold nanoparticles and its biological applications. The production of fucoidanase was optimized using Streptomyces sp. The medium components were selected in accordance with the Plackett-Burman design and were further optimized via response surface methodology. The fucoidanase was statistically optimized with the most significant factors, namely wheat bran 3.3441 g/L, kelp powder 0.7041 g/L, and NaCl 0.8807 g/L, respectively. The biosynthesized gold nanoparticles were determined by UV-vis spectroscopy and were further characterized by X-ray diffraction analysis, Fourier transform infrared spectroscopy, field emission scanning electron microscopy, energy dispersive X-ray analysis, and high-resolution transmission electron microscopy. Furthermore, the biosynthesized gold nanoparticles exhibited a dose-dependent cytotoxicity against HeLa cells and the inhibitory concentration (IC50) was found to be 350 µg/mL at 24 h and 250 µg/mL at 48 h. Therefore, the production of novel fucoidanase for the green synthesis of gold nanoparticles has comparatively rapid, less expensive and wide application to anticancer therapy in modern medicine.

  16. Isolation and characterization of a proteinaceous α-amylase inhibitor AAI-CC5 from Streptomyces sp. CC5, and its gene cloning and expression.

    PubMed

    Sun, Zhibin; Lu, Weihao; Liu, Pingping; Wang, Hui; Huang, Yan; Zhao, Yuguo; Kong, Yi; Cui, Zhongli

    2015-02-01

    An α-amylase inhibitor producing Streptomyces sp. strain CC5 was isolated from soil. A proteinaceous α-amylase inhibitor AAI-CC5 was purified from strain CC5. AAI-CC5 specifically inhibited mammalian α-amylases. The molecular weight of the inhibitor was determined to be 8,212 Da by MALDI-TOF Mass Spectrum. The N-terminal 15 amino acid residues of the purified AAI-CC5 were DTGSPAPECVEYFQS, which is dissimilar to other reported proteinaceous α-amylase inhibitors. AAI-CC5 is a pH insensitive and heat-stable protein, and cannot be hydrolysed by trypsin. AAI-CC5 was cloned and expressed in Escherichia coli BL21 (DE3) with a hexa-histidine tag on the C terminal. AAI-CC5 shared 82 % identity with Parvulustat. The recombinant α-amylase inhibitor was purified to homogeneity by one-step affinity chromatography using Ni(2+)-NTA resin with molecular mass of 9,404 Da. Steady state kinetics studies of α-amylase and the inhibitor revealed an irreversible, non-competitive inhibition mechanism with IC50 and Ki value of 6.43 ×1 10(-11) and 4.45 × 10(-11) M respectively. These results suggest this novel α-amylase inhibitor possessed powerful inhibitory activity for α-amylase, and it may be a candidate in research of diabetes therapy and obesity treatment.

  17. Marine Streptomyces sp. derived antimycin analogues suppress HeLa cells via depletion HPV E6/E7 mediated by ROS-dependent ubiquitin–proteasome system

    PubMed Central

    Zhang, Weiyi; Che, Qian; Tan, Hongsheng; Qi, Xin; Li, Jing; Li, Dehai; Gu, Qianqun; Zhu, Tianjiao; Liu, Ming

    2017-01-01

    Four new antimycin alkaloids (1–4) and six related known analogs (5–10) were isolated from the culture of a marine derived Streptomyces sp. THS-55, and their structures were elucidated by extensive spectroscopic analysis. All of the compounds exhibited potent cytotoxicity in vitro against HPV-transformed HeLa cell line. Among them, compounds 6–7 were derived as natural products for the first time, and compound 5 (NADA) showed the highest potency. NADA inhibited the proliferation, arrested cell cycle distribution, and triggered apoptosis in HeLa cancer cells. Our molecular mechanic studies revealed NADA degraded the levels of E6/E7 oncoproteins through ROS-mediated ubiquitin-dependent proteasome system activation. This is the first report that demonstrates antimycin alkaloids analogue induces the degradation of high-risk HPV E6/E7 oncoproteins and finally induces apoptosis in cervical cancer cells. The present work suggested that these analogues could serve as lead compounds for the development of HPV-infected cervical cancer therapeutic agents, as well as research tools for the study of E6/E7 functions. PMID:28176847

  18. Bioweathering and biotransformation of granitic rock minerals by actinomycetes.

    PubMed

    Abdulla, Hesham

    2009-11-01

    Actinomycetes inhabiting granitic rocks at St. Katherine, Egypt were investigated for their bioweathering potential. Actinomycete counts ranged between 174 and 360 colony forming units per gram. Counts were positively correlated to rock porosity (r = 0.65) and negatively correlated to rock salinity (r = -0.56). Sixty-six actinomycete isolates originating from rocks could be assigned into eight genera, with a high frequency of Nocardioides and Streptomyces. Organic acids were produced by 97% of the isolates. Strains belonging to Actinopolyspora, Actinomadura, Kitasatospora, Nocardioides, and Kibdelosporangium showed the highest acid production indices. Representatives from all eight genera could precipitate metals Cu, Fe, Zn, Cd, and Ag up to concentrations of 2.5 mM each. An actinomycete consortium of two Nocardioides strains and one Kibdelosporangium strain was studied for its potential to cause rock weathering in batch experiments. Results indicated a high ability of the consortium to leach the metals Cu, Zn, and Fe up to 2.6-, 2.1-, and 1.3-fold, respectively, compared to the control after 4 weeks. The pH significantly decreased after 1 week, which was parallel to an increased release of phosphate and sulfate reaching a 2.2- and 2.5-fold increase, respectively, compared to control. Highly significant weight loss (p = 0.005) was achieved by the consortium, indicating a potential multiple role of actinomycetes in weathering by acid production, metal leaching, and solubilization of phosphate and sulfate. This study emphasizes the diverse and unique abilities of actinomycetes inhabiting rock surfaces which could be of potential biotechnological applications, such as in the bioremediation of metal-contaminated environments and metal biorecovery.

  19. Characterization and structure elucidation of antibacterial compound of Streptomyces sp. ECR77 isolated from east coast of India.

    PubMed

    Thirumurugan, D; Vijayakumar, R

    2015-05-01

    Forty marine actinobacteria were isolated from the sediments of east coast (Bay of Bengal) region of Tamilnadu, India. Morphologically distinct colonies were primarily tested against fish pathogenic bacteria such as Vibrio cholerae, V. parahaemolyticus, V. alginolyticus, Pseudomonas fluorescens and Aeromonas hydrophila by cross-streak plate method. The secondary metabolites produced by the highly potential strain cultured on starch casein broth were extracted separately with various solvents such as alcohol, ethyl acetate, methanol, petroleum ether and chloroform. The antibacterial assay of the bioactive compounds was tested against the fish pathogenic bacteria by well diffusion method. Of the various solvents used, the ethyl acetate extract of the isolate had good antibacterial activity. The potential strain was identified as Streptomyces labedae by phenotypic, 16S rRNA gene sequence and phylogenetic analysis. Purification of the biologically active compounds by column chromatography led to isolation of 27 fractions. The biologically active fraction was re-chromatographed on a silica gel column to obtain a single active compound, namely N-isopentyltridecanamide. The structure of the compounds was elucidated on the basis of ultra violet, Fourier transform infrared and nuclear magnetic resonance spectra.

  20. Characterization of a PA14 domain-containing galactofuranose-specific β-d-galactofuranosidase from Streptomyces sp.

    PubMed

    Matsunaga, Emiko; Higuchi, Yujiro; Mori, Kazuki; Yairo, Nao; Toyota, Saki; Oka, Takuji; Tashiro, Kosuke; Takegawa, Kaoru

    2017-03-20

    As a constituent of polysaccharides and glycoconjugates, β-d-galactofuranose (Galf) exists in several pathogenic microorganisms. Although we recently identified a β-d-galactofuranosidase (Galf-ase) gene, ORF1110, in the Streptomyces strain JHA19, very little is known about the Galf-ase gene. Here, we characterized a strain, named JHA26, in the culture supernatant of which exhibited Galf-ase activity for 4-nitrophenyl β-d-galactofuranoside (pNP-β-d-Galf) as a substrate. Draft genome sequencing of the JHA26 strain revealed a putative gene, termed ORF0643, that encodes Galf-ase containing a PA14 domain, which is thought to function in substrate recognition. The recombinant protein expressed in Escherichia coli showed the Galf-specific Galf-ase activity and also released galactose residue of the polysaccharide galactomannan prepared from Aspergillus fumigatus, suggesting that this enzyme is an exo-type Galf-ase. BLAST searches using the amino acid sequences of ORF0643 and ORF1110 Galf-ases revealed two types of Galf-ases in Actinobacteria, suggesting that Galf-specific Galf-ases may exhibit discrete substrate specificities.

  1. Actinomycetes with antimicrobial activity isolated from paper wasp (Hymenoptera: Vespidae: Polistinae) nests.

    PubMed

    Madden, Anne A; Grassetti, Andrew; Soriano, Jonathan-Andrew N; Starks, Philip T

    2013-08-01

    Actinomycetes-a group of antimicrobial producing bacteria-have been successfully cultured and characterized from the nest material of diverse arthropods. Some are symbionts that produce antimicrobial chemicals found to protect nest brood and resources from pathogenic microbes. Others have no known fitness relationship with their associated insects, but have been found to produce antimicrobials in vitro. Consequently, insect nest material is being investigated as a new source of novel antimicrobial producing actinomycetes, which could be harnessed for therapeutic potential. To extend studies of actinomycete-insect associations beyond soil-substrate dwelling insects and wood boring excavators, we conducted a preliminary assessment of the actinomycetes within the nests of the paper wasp, Polistes dominulus (Christ). We found that actinomycetes were readily cultured from nest material across multiple invasive P. dominulus populations-including members of the genera Streptomyces, Micromonospora, and Actinoplanes. Thirty of these isolates were assayed for antimicrobial activity against the challenge bacteria Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Serratia marcescens, and Bacillus subtilis. Sixty percent of isolates inhibited the growth of at least one challenge strain. This study provides the first assessment of bacteria associated with nests of P. dominulus, and the first record of antimicrobial producing actinomycetes isolated from social wasps. We provide a new system to explore nest associated actinomycetes from a ubiquitous and cosmopolitan group of insects.

  2. Isolation and identification of actinomycetes for production of novel extracellular glutaminase free L-asparaginase.

    PubMed

    Saxena, Akansha; Upadhyay, Ramraj; Kango, Naveen

    2015-12-01

    Over the recent years glutaminase free L-asparaginase has gained more importance due to better therapeutic properties for treatment of acute lymphoblastic leukemia. Actinomycetes are known for L-asparaginase activity. In the current study, 80 actinomycetes were isolated from various soil habitats by serial dilution technique. Presence of L-asparaginase was investigated in a total of 240 actinomycetes by tubed agar method using modified M-9 medium. A total of 165 actinomycetes were found positive for L-asparaginase activity. Among these, 57 actinomycetes producing larger zones of L-asparagine hydrolysis were further screened for their capacity to produce glutaminase-free L-asparaginase. Four L-glutaminase-free actinomycetes were found to be potential L-asparaginase producers. These actinomycetes were identified as Streptomyces cyaneus (SAP 1287, CFS 1560), S. exfoliates (CFS 1557) and S. phaeochromogenes (GS 1573) on the basis of morphological and biochemical identification studies. Maximum L-asparaginase activity (19.2 Uml(-1)) was observed in culture filtrate of S. phaeochromogenes under submerged fermentation. Results indicate that S. phaeochromogenes could be a potential source of glutaminase free L-asparaginase for commercial purpose. To the best of our knowledge, this is the first report on production of glutaminase free L-asparaginase from S. cyaneus, S. exfoliatus and S. phaeochromogenes.

  3. A novel taxonomic marker that discriminates between morphologically complex actinomycetes.

    PubMed

    Girard, Geneviève; Traag, Bjørn A; Sangal, Vartul; Mascini, Nadine; Hoskisson, Paul A; Goodfellow, Michael; van Wezel, Gilles P

    2013-10-23

    In the era when large whole genome bacterial datasets are generated routinely, rapid and accurate molecular systematics is becoming increasingly important. However, 16S ribosomal RNA sequencing does not always offer sufficient resolution to discriminate between closely related genera. The SsgA-like proteins are developmental regulatory proteins in sporulating actinomycetes, whereby SsgB actively recruits FtsZ during sporulation-specific cell division. Here, we present a novel method to classify actinomycetes, based on the extraordinary way the SsgA and SsgB proteins are conserved. The almost complete conservation of the SsgB amino acid (aa) sequence between members of the same genus and its high divergence between even closely related genera provides high-quality data for the classification of morphologically complex actinomycetes. Our analysis validates Kitasatospora as a sister genus to Streptomyces in the family Streptomycetaceae and suggests that Micromonospora, Salinispora and Verrucosispora may represent different clades of the same genus. It is also apparent that the aa sequence of SsgA is an accurate determinant for the ability of streptomycetes to produce submerged spores, dividing the phylogenetic tree of streptomycetes into liquid-culture sporulation and no liquid-culture sporulation branches. A new phylogenetic tree of industrially relevant actinomycetes is presented and compared with that based on 16S rRNA sequences.

  4. Actinomycetes for marine drug discovery isolated from mangrove soils and plants in China.

    PubMed

    Hong, Kui; Gao, An-Hui; Xie, Qing-Yi; Gao, Hao; Zhuang, Ling; Lin, Hai-Peng; Yu, Hai-Ping; Li, Jia; Yao, Xin-Sheng; Goodfellow, Michael; Ruan, Ji-Sheng

    2009-01-01

    The mangrove ecosystem is a largely unexplored source for actinomycetes with the potential to produce biologically active secondary metabolites. Consequently, we set out to isolate, characterize and screen actinomycetes from soil and plant material collected from eight mangrove sites in China. Over 2,000 actinomycetes were isolated and of these approximately 20%, 5%, and 10% inhibited the growth of Human Colon Tumor 116 cells, Candida albicans and Staphylococcus aureus, respectively, while 3% inhibited protein tyrosine phosphatase 1B (PTP1B), a protein related to diabetes. In addition, nine isolates inhibited aurora kinase A, an anti-cancer related protein, and three inhibited caspase 3, a protein related to neurodegenerative diseases. Representative bioactive isolates were characterized using genotypic and phenotypic procedures and classified to thirteen genera, notably to the genera Micromonospora and Streptomyces. Actinomycetes showing cytotoxic activity were assigned to seven genera whereas only Micromonospora and Streptomyces strains showed anti-PTP1B activity. We conclude that actinomycetes isolated from mangrove habitats are a potentially rich source for the discovery of anti-infection and anti-tumor compounds, and of agents for treating neurodegenerative diseases and diabetes.

  5. Extracellular production of the oncolytic enzyme, L-asparaginase, by newly isolated Streptomyces sp. strain NEAE-95 as potential microbial cell factories: optimization of culture conditions using response surface methodology.

    PubMed

    El-Naggar, Noura E-A

    2015-01-01

    L-asparaginase is an effective anti-neoplastic agent used in chemotherapy of acute lymphoblastic leukemia. One hundred and thirty actinomycete isolates were isolated from several soil samples collected from different localities in Egypt. All these isolates were purified and evaluated for their ability to produce L-asparaginase activity. Among them, strain NEAE-95 was selected and identified as Streptomyces parvus strain NEAE-95 based on morphological, cultural, physiological characteristics and 16S rRNA sequence. The sequence was deposited in the NCBI GenBank database under accession number KJ200341. L-asparaginase production by Streptomyces parvus NEAE-95 was optimized in shake flask culture. The Plackett-Burman statistical design was used for initial screening of sixteen factors for their significance on L-asparaginase production. Among the variables screened, incubation time, L-asparagine and yeast extract had significant effects on L-asparaginase production. The levels of these significant variables and their interaction effects were optimized by Box-Behnken statistical design. As a result, the maximal L-asparaginase production was achieved at the following fermentation conditions: g/L (dextrose 2, starch 20, L-asparagine 14, KNO3 2, yeast extract 2, K2HPO4 2, MgSO4.7H2O 0.1, NaCl 0.1, FeSO4.7H2O 0.01), pH 7, temperature 30°C, agitation speed 200 rpm, inoculum size 2%, v/v and incubation time 8 days.

  6. Geosmin, an Earthy-Smelling Substance Isolated from Actinomycetes

    PubMed Central

    Gerber, N. N.; Lechevalier, H. A.

    1965-01-01

    Geosmin, an earthy-smelling substance, has been isolated from several actinomycetes. Production of 1 mg per liter of whole broth was obtained from Streptomyces griseus LP-16. After preliminary separations, pure geosmin was isolated in milligram amounts by gas chromatography. Geosmin is a neutral oil, with an approximate boiling point of 270 C, which contains carbon and hydrogen, but no nitrogen. It undergoes a reaction with acid to give odorless argosmin, a neutral oil, with an approximate boiling point of 230 C, which contains only carbon and hydrogen. Specific rotation and ultraviolet- and infrared-absorbtion spectra were determined for both. PMID:5866039

  7. Study of the effects of urban organic residues on the distribution of culturable actinomycetes in a Tunisian agricultural soil.

    PubMed

    Mokni-Tlili, Sonia; Jaoua, Leila; Murano, Fumio; Jedidi, Naceur; Hassen, Abdennaceur

    2009-05-01

    The main objective of this investigation was to identify a collection of actinomycetes isolates and to study the influence of amendment [municipal solid waste compost (MSWC) and farmyard manure (FM)] on their distribution in agricultural soil. For this purpose, a phenotypic and molecular characterization of 226 isolates collected from soil (with and without amendment) and 55 isolates from MSWC and FM was developed. The phenotypic study showed that the majority of strains isolated belong to the genus Streptomyces. By using the 16S rDNA polymerase chain reaction-restriction fragment length polymorphism method (restriction digest using six enzymes AluI, HhaI, MspI, TaqI, RsaI and HaeIII), two clusters were found: Streptomyces, dominant genus and Amycolatopsis, followed by Nocardioides. This result agreed with phylogeny revealed by 16S rDNA sequencing. The number of these actinomycetes in soil increased with FM or MSWC application. The studied soil is a potential source for isolation of actinomycetes, especially Streptomyces, and the application of organic amendment to the soil appeared to have an impact on the diversity of actinomycetes. Amendment of the soil with MSWC and FM significantly increased the number of actinomycetes due to the contribution of bacteria originally contained in biowastes and/or by stimulation of the endogenous soil micro-organisms.

  8. Catalytic properties of an organic solvent-resistant tyrosinase from Streptomyces sp. REN-21 and its high-level production in E. coli.

    PubMed

    Ito, Masaaki; Inouye, Kuniyo

    2005-10-01

    An organic solvent-resistant tyrosinase (OSRT) from Streptomyces sp. REN-21 is a unique enzyme showing high activity in the presence of organic solvents. The OSRT-catalyzed oxidation of monophenols such as tyrosine-containing peptides and proteins was examined. The catalytic properties of OSRT were compared with those of mushroom tyrosinase. OSRT was shown to oxidize Gly-l-Tyr most effectively among four peptide substrates tested. On the other hand, mushroom tyrosinase showed the highest activity toward l-Tyr-Gly under the condition of 1 mM substrate. OSRT oxidized several proteins, including casein and hemoglobin, with relatively higher activity compared with mushroom tyrosinase under the condition of 1% (w/v) substrate. Thus, it was clarified that the catalytic properties of OSRT toward tyrosine-containing peptides and proteins are different from those of mushroom tyrosinase under these conditions. The OSRT-encoding gene operon was cloned, and found to consist of two genes, designated ORF-OSRT and ORF-393. The former encodes apo-OSRT, and the latter encodes the putative activator protein of apo-OSRT. A binuclear copper-binding site (type-3 copper site) characteristic of tyrosinases is contained in the deduced amino acid sequence for apo-OSRT. A high-level production system for the OSRT was constructed using pET20b(+) and Escherichia coli BL21(DE3)pLysS. Approximately 54 mg of active OSRT was synthesized in a 1-liter broth culture by this system. The properties of the recombinant OSRT were similar to those of the wild-type enzyme. In conclusion, we succeeded in constructing a high-level production system for OSRT.

  9. Targeted genome editing in the rare actinomycete Actinoplanes sp. SE50/110 by using the CRISPR/Cas9 System.

    PubMed

    Wolf, Timo; Gren, Tetiana; Thieme, Eric; Wibberg, Daniel; Zemke, Till; Pühler, Alfred; Kalinowski, Jörn

    2016-08-10

    The application of genome editing technologies, like CRISPR/Cas9 for industrially relevant microorganisms, is becoming increasingly important. Compared to other methods of genetic engineering the decisive factor is that CRISPR/Cas9 is relatively easy to apply and thus time and effort can be significantly reduced in organisms, which are otherwise genetically difficult to access. Because of its many advantages and opportunities, we adopted the CRISPR/Cas9 technology for Actinoplanes sp. SE50/110, the producer of the diabetes type II drug acarbose. The functionality of genome editing was successfully shown by the scarless and antibiotic marker-free deletion of the gene encoding the tyrosinase MelC, which catalyzes the formation of the dark pigment eumelanin in the wild type strain. The generated ΔmelC2 mutant of Actinoplanes sp. SE50/110 no longer produces this pigment and therefore the supernatant does not darken. Furthermore, it was shown that the plasmid containing the gene for the Cas9 protein was removed by increasing the temperature due to its temperature-sensitive replication. The precision of the intended mutation was proven and possible off-target effects caused by the genome editing system were ruled out by genome sequencing of several mutants.

  10. The Madeira Archipelago As a Significant Source of Marine-Derived Actinomycete Diversity with Anticancer and Antimicrobial Potential.

    PubMed

    Prieto-Davó, Alejandra; Dias, Tiago; Gomes, Sofia E; Rodrigues, Sara; Parera-Valadez, Yessica; Borralho, Pedro M; Pereira, Florbela; Rodrigues, Cecilia M P; Santos-Sanches, Ilda; Gaudêncio, Susana P

    2016-01-01

    Marine-derived actinomycetes have demonstrated an ability to produce novel compounds with medically relevant biological activity. Studying the diversity and biogeographical patterns of marine actinomycetes offers an opportunity to identify genera that are under environmental pressures, which may drive adaptations that yield specific biosynthetic capabilities. The present study describes research efforts to explore regions of the Atlantic Ocean, specifically around the Madeira Archipelago, where knowledge of the indigenous actinomycete diversity is scarce. A total of 400 actinomycetes were isolated, sequenced, and screened for antimicrobial and anticancer activities. The three most abundant genera identified were Streptomyces, Actinomadura, and Micromonospora. Phylogenetic analyses of the marine OTUs isolated indicated that the Madeira Archipelago is a new source of actinomycetes adapted to life in the ocean. Phylogenetic differences between offshore (>100 m from shore) and nearshore (< 100 m from shore) populations illustrates the importance of sampling offshore in order to isolate new and diverse bacterial strains. Novel phylotypes from chemically rich marine actinomycete groups like MAR4 and the genus Salinispora were isolated. Anticancer and antimicrobial assays identified Streptomyces, Micromonospora, and Salinispora as the most biologically active genera. This study illustrates the importance of bioprospecting efforts at unexplored regions of the ocean to recover bacterial strains with the potential to produce novel and interesting chemistry.

  11. The Madeira Archipelago As a Significant Source of Marine-Derived Actinomycete Diversity with Anticancer and Antimicrobial Potential

    PubMed Central

    Prieto-Davó, Alejandra; Dias, Tiago; Gomes, Sofia E.; Rodrigues, Sara; Parera-Valadez, Yessica; Borralho, Pedro M.; Pereira, Florbela; Rodrigues, Cecilia M. P.; Santos-Sanches, Ilda; Gaudêncio, Susana P.

    2016-01-01

    Marine-derived actinomycetes have demonstrated an ability to produce novel compounds with medically relevant biological activity. Studying the diversity and biogeographical patterns of marine actinomycetes offers an opportunity to identify genera that are under environmental pressures, which may drive adaptations that yield specific biosynthetic capabilities. The present study describes research efforts to explore regions of the Atlantic Ocean, specifically around the Madeira Archipelago, where knowledge of the indigenous actinomycete diversity is scarce. A total of 400 actinomycetes were isolated, sequenced, and screened for antimicrobial and anticancer activities. The three most abundant genera identified were Streptomyces, Actinomadura, and Micromonospora. Phylogenetic analyses of the marine OTUs isolated indicated that the Madeira Archipelago is a new source of actinomycetes adapted to life in the ocean. Phylogenetic differences between offshore (>100 m from shore) and nearshore (< 100 m from shore) populations illustrates the importance of sampling offshore in order to isolate new and diverse bacterial strains. Novel phylotypes from chemically rich marine actinomycete groups like MAR4 and the genus Salinispora were isolated. Anticancer and antimicrobial assays identified Streptomyces, Micromonospora, and Salinispora as the most biologically active genera. This study illustrates the importance of bioprospecting efforts at unexplored regions of the ocean to recover bacterial strains with the potential to produce novel and interesting chemistry. PMID:27774089

  12. Anti-phytopathogen potential of endophytic actinobacteria isolated from tomato plants (Lycopersicon esculentum) in southern Brazil, and characterization of Streptomyces sp. R18(6), a potential biocontrol agent.

    PubMed

    de Oliveira, Margaroni Fialho; da Silva, Mariana Germano; Van Der Sand, Sueli T

    2010-09-01

    Tomato plants (Lycopersicon esculentum) are highly susceptible to phytopathogen attack. The resulting intensive application of pesticides on tomato crops can affect the environment and health of humans and animals. The objective of this study was to select potential biocontrol agents among actinobacteria from tomato plants, in a search for alternative phytopathogen control. We evaluated 70 endophytic actinobacteria isolated from tomato plants in southern Brazil, testing their antimicrobial activity, siderophore production, indoleacetic acid production, and phosphate solubility. The actinomycete isolate with the highest antimicrobial potential was selected using the agar-well diffusion method, in order to optimize conditions for the production of compounds with antimicrobial activity. For this study, six growth media (starch casein-SC, ISP2, Bennett's, Sahin, Czapek-Dox, and TSB), three temperatures (25 degrees C, 30 degrees C, and 35 degrees C) and different pH were tested. Of the actinobacteria tested, 88.6% showed antimicrobial activity against at least one phytopathogen, 72.1% showed a positive reaction for indoleacetic acid production, 86.8% produced siderophores and 16.2% showed a positive reaction for phosphate solubility. Isolate R18(6) was selected due to its antagonistic activity against all phytopathogenic microorganisms tested in this study. The best conditions for production were observed in the SC medium, at 30 degrees C and pH 7.0. The isolate R18(6) showed close biochemical and genetic similarity to Streptomyces pluricolorescens.

  13. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

    PubMed

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  14. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants

    PubMed Central

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10–32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  15. Screening of Rhizospheric Actinomycetes for Various In-vitro and In-vivo Plant Growth Promoting (PGP) Traits and for Agroactive Compounds

    PubMed Central

    Anwar, Sumaira; Ali, Basharat; Sajid, Imran

    2016-01-01

    In this study 98 rhizospheric actinomycetes were isolated from different wheat and tomato fields, Punjab, Pakistan. The isolates were characterized morphologically, biochemically, and genetically and were subjected to a comprehensive in vitro screening for various plant growth promoting (PGP) traits. About 30% of the isolates screened were found to be the promising PGP rhizobacteria (PGPRs), which exhibited maximum genetic similarity (up to 98–99%) with different species of the genus Streptomyces by using16S rRNA gene sequencing. The most active indole acetic acid (IAA) producer Streptomyces nobilis WA-3, Streptomyces Kunmingenesis WC-3, and Streptomyces enissocaesilis TA-3 produce 79.5, 79.23, and 69.26 μg/ml IAA respectively at 500 μg/ml L-tryptophan. The highest concentration of soluble phosphate was produced by Streptomyces sp. WA-1 (72.13 mg/100 ml) and S. djakartensis TB-4 (70.36 mg/100 ml). All rhizobacterial isolates were positive for siderophore, ammonia, and hydrogen cyanide production. Strain S. mutabilis WD-3 showed highest concentration of ACC-deaminase (1.9 mmol /l). For in-vivo screening, seed germination, and plant growth experiment were conducted by inoculating wheat (Triticum aestivum) seeds with the six selected isolates. Significant increases in shoot length was observed with S. nobilis WA-3 (65%), increased root length was recorded in case of S. nobilis WA-3 (81%) as compared to water treated control plants. Maximum increases in plant fresh weight were recorded with S. nobilis WA-3 (84%), increased plant dry weight was recorded in case of S. nobilis WA-3 (85%) as compared to water treated control plants. In case of number of leaves, significant increase was recorded with S. nobilis WA-3 (27%) and significant increase in case of number of roots were recorded in case of strain S. nobilis WA-3 (30%) as compared to control plants. Over all the study revealed that these rhizospheric PGP Streptomyces are good candidates to be developed as

  16. Streptomyces phospholipase D cloning and production.

    PubMed

    Nakazawa, Yozo

    2012-01-01

    The transphosphatidylation catalytic ability of phospholipase D (PLD, EC 3.1.4.4) is a powerful biochemical tool for the acquisition of rare phospholipids (PLs), e.g., phosphatidylglycerol (PG) and phosphatidylserine (PS), and artificial phospholipids, which do not occur in nature. Specifically, actinomycete PLD recognizes not only the alcohols (i.e., glycerol or serine) corresponding to the polar head groups of natural PLs, but also secondary alcohols, aromatic alcohols, saccharides, N-heterocyclic alcohols, and vitamins as acceptors. Therefore, actinomycete PLD is a valuable enzyme in food, cosmetics, fine chemical and pharmaceutical industries. Here, we describe a protocol for the screening for PLD-producing microorganisms, several PLD assays and methods of PLD production-purification and the strategy of cloning of the Streptomyces PLD gene.

  17. Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82

    PubMed Central

    El-Naggar, Noura El-Ahmady; Deraz, Sahar F.; Soliman, Hoda M.; El-Deeb, Nehal M.; El-Ewasy, Sara M.

    2016-01-01

    L-asparaginase is an important enzyme as therapeutic agents used in combination with other drugs in the treatment of acute lymphoblastic leukemia. A newly isolated actinomycetes strain, Streptomyces sp. NEAE-82, was potentially producing extracellular L-asparaginase, it was identified as Streptomyces fradiae NEAE-82, sequencing product was deposited in the GenBank database under accession number KJ467538. L-asparaginase was purified from the crude enzyme using ammonium sulfate precipitation, dialysis and ion exchange chromatography using DEAE Sepharose CL-6B. Further the kinetic studies of purified enzyme were carried out. The optimum pH, temperature and incubation time for maximum L-asparaginase activity were found to be 8.5, 40 °C and 30 min, respectively. The optimum substrate concentration was found to be 0.06 M. The Km and Vmax of the enzyme were 0.01007 M and 95.08 Uml−1min−1, respectively. The half-life time (T1/2) was 184.91 min at 50 °С, while being 179.53 min at 60 °С. The molecular weight of the subunits of L-asparaginase was found to be approximately 53 kDa by SDS–PAGE analysis. The purified L-asparaginase showed a final specific activity of 30.636 U/mg protein and was purified 3.338-fold. The present work for the first time reported more information in the production, purification and characterization of L-asparaginase produced by newly isolated actinomycetes Streptomyces fradiae NEAE-82. PMID:27605431

  18. A mixed community of actinomycetes produce multiple antibiotics for the fungus farming ant Acromyrmex octospinosus

    PubMed Central

    2010-01-01

    Background Attine ants live in an intensely studied tripartite mutualism with the fungus Leucoagaricus gongylophorus, which provides food to the ants, and with antibiotic-producing actinomycete bacteria. One hypothesis suggests that bacteria from the genus Pseudonocardia are the sole, co-evolved mutualists of attine ants and are transmitted vertically by the queens. A recent study identified a Pseudonocardia-produced antifungal, named dentigerumycin, associated with the lower attine Apterostigma dentigerum consistent with the idea that co-evolved Pseudonocardia make novel antibiotics. An alternative possibility is that attine ants sample actinomycete bacteria from the soil, selecting and maintaining those species that make useful antibiotics. Consistent with this idea, a Streptomyces species associated with the higher attine Acromyrmex octospinosus was recently shown to produce the well-known antifungal candicidin. Candicidin production is widespread in environmental isolates of Streptomyces, so this could either be an environmental contaminant or evidence of recruitment of useful actinomycetes from the environment. It should be noted that the two possibilities for actinomycete acquisition are not necessarily mutually exclusive. Results In order to test these possibilities we isolated bacteria from a geographically distinct population of A. octospinosus and identified a candicidin-producing Streptomyces species, which suggests that they are common mutualists of attine ants, most probably recruited from the environment. We also identified a Pseudonocardia species in the same ant colony that produces an unusual polyene antifungal, providing evidence for co-evolution of Pseudonocardia with A. octospinosus. Conclusions Our results show that a combination of co-evolution and environmental sampling results in the diversity of actinomycete symbionts and antibiotics associated with attine ants. PMID:20796277

  19. Streptomyces rochei ACTA1551, an Indigenous Greek Isolate Studied as a Potential Biocontrol Agent against Fusarium oxysporum f.sp. lycopersici

    PubMed Central

    Kanini, Grammatiki S.; Katsifas, Efstathios A.; Savvides, Alexandros L.; Karagouni, Amalia D.

    2013-01-01

    Many studies have shown that several Greek ecosystems inhabit very interesting bacteria with biotechnological properties. Therefore Streptomyces isolates from diverse Greek habitats were selected for their antifungal activity against the common phytopathogenic fungus Fusarium oxysporum. The isolate encoded ACTA1551, member of Streptomyces genus, could strongly suppress the fungal growth when examined in antagonistic bioassays in vitro. The isolate was found phylogenetically relative to Streptomyces rochei after analyzing its 16S rDNA sequence. The influence of different environmental conditions, such as medium composition, temperature, and pH on the expression of the antifungal activity was thoroughly examined. Streptomyces rochei ACTA1551 was able to protect tomato seeds from F. oxysporum infection in vivo while it was shown to promote the growth of tomato plants when the pathogen was absent. In an initial effort towards the elucidation of the biochemical and physiological nature of ACTA1551 antifungal activity, extracts from solid streptomycete cultures under antagonistic or/and not antagonistic conditions were concentrated and fractionated. The metabolites involved in the antagonistic action of the isolate showed to be more than one and produced independently of the presence of the pathogen. The above observations could support the application of Streptomyces rochei ACTA1551 as biocontrol agent against F. oxysporum. PMID:23762841

  20. Ecological and Taxonomic Features of Actinomycetal Complexes in Soils of the Lake Elton Basin

    NASA Astrophysics Data System (ADS)

    Zenova, G. M.; Dubrova, M. S.; Kuznetsova, A. I.; Gracheva, T. A.; Manucharova, N. A.; Zvyagintsev, D. G.

    2016-02-01

    In the sor (playa) solonchaks of chloride and sulfate-chloride salinity (the content of readily soluble salts is 0.9-1.0%) in the delta of the Khara River discharging into Lake Elton, the number of mycelial actinobacteria (actinomycetes) is low ((2-3) × 103 CFU/g of soil). At a distance from the water's edge, these soils are substituted for the light chestnut ones, for which an elevated number of actinomycetes (an order of magnitude higher than in the sor solonchaks) and a wider generic spectrum are characteristic. The actinomycetal complex is included the Streptomyces and Micromonospora genera, whereas in the sor solonchaks around the lake, representatives of Micromonospora were not found.

  1. Salininema proteolyticum gen. nov., sp. nov., a halophilic rare actinomycete isolated from wetland soil, and emended description of the family Glycomycetaceae.

    PubMed

    Nikou, Mahdi Moshtaghi; Ramezani, Mohaddaseh; Amoozegar, Mohammad Ali; Rasouli, Mehrnoush; Fazeli, Seyed Abolhassan Shahzadeh; Schumann, Peter; de la Haba, Rafael R; Ventosa, Antonio

    2015-10-01

    A Gram-stain-positive actinobacterial strain, Miq-4T, was isolated from soil around Meighan wetland in the centre of Iran. Strain Miq-4T was strictly aerobic, catalase- and oxidase-positive. The isolate grew in the presence of 3–15 % (w/v) NaCl, at 20–40 °C and pH 6.0–11.0. The optimum NaCl, temperature and pH for growth were 7.0 %, 30 °C and 7.0–8.5, respectively. The cell wall of strain Miq-4T contained meso-diaminopimelic acid as the diamino acid and glucose and ribose as the whole-cell sugars. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Strain Miq-4T synthesized cellular fatty acids of anteiso- and iso-branched types, including anteiso-C17 : 0, anteiso- C15 : 0 and iso-C16 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 68.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences and characteristic patterns of 16S rRNA gene signature nucleotides revealed that strain Miq-4T belongs to the family Glycomycetaceae and showed the closest phylogenetic similarity with Haloglycomyces albus YIM 92370T (94.1 % 16S rRNA gene sequence similarity). On the basis of phylogenetic analysis and phenotypic and chemotaxonomic characteristics, strain Miq-4T represents a novel species of a new genus in the family Glycomycetaceae, for which the name Salininema proteoliyticum gen. nov., sp. nov. is proposed. The type strain of the type species is Miq-4T ( = IBRC-M 10908T = LMG 28391T). An emended description of the family Glycomycetaceae is also proposed in order to include features of the new genus.

  2. Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea.

    PubMed

    Sreevidya, M; Gopalakrishnan, S; Kudapa, H; Varshney, R K

    2016-01-01

    The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20°C to 40°C, pH range of 7-11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea.

  3. Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea

    PubMed Central

    Sreevidya, M.; Gopalakrishnan, S.; Kudapa, H.; Varshney, R.K.

    2016-01-01

    The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20 °C to 40 °C, pH range of 7–11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea. PMID:26887230

  4. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    PubMed Central

    Bernal, Milagro García; Campa-Córdova, Ángel Isidro; Saucedo, Pedro Enrique; González, Marlen Casanova; Marrero, Ricardo Medina; Mazón-Suástegui, José Manuel

    2015-01-01

    Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4) among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world. PMID:27047067

  5. [Isolation and antimicrobial activities of actinomycetes from vermicompost].

    PubMed

    Wang, Xue-jun; Yan, Shuang-lin; Min, Chang-li; Yang, Yan

    2015-02-01

    In this paper, actinomycetes were isolated from vermicompost by tablet coating method. Antimicrobial activities of actinomycetes were measured by the agar block method. Strains with high activity were identified based on morphology and biochemical characteristics, as well as 16S rDNA gene sequence analysis. The results showed that 26 strains of actinomycetes were isolated, 16 of them had antimicrobial activities to the test strains which accounts for 61.54% of all strains. Among the 16 strains, the strain QYF12 and QYF22 had higher antimicrobial activity to Micrococcus luteus, with a formed inhibition zone of 27 mm and 31 mm, respectively. While the strain QYF26 had higher antimicrobial activity to Bacillus subtilis, and the inhibition zone diameter was 21 mm. Based on the identification of strains with high activity, the strain QYF12 was identified as Streptomyces chartreusis, the strain QYF22 was S. ossamyceticus and the strain QYF26 was S. gancidicus. This study provided a theoretical basis for further separate antibacterial product used for biological control.

  6. Artificial chromosomes to explore and to exploit biosynthetic capabilities of actinomycetes.

    PubMed

    Alduina, Rosa; Gallo, Giuseppe

    2012-01-01

    Actinomycetes are an important source of biologically active compounds, like antibiotics, antitumor agents, and immunosuppressors. Genome sequencing is revealing that this class of microorganisms has larger genomes relative to other bacteria and uses a considerable fraction of its coding capacity (5-10%) for the production of mostly cryptic secondary metabolites. To access actinomycetes biosynthetic capabilities or to improve the pharmacokinetic properties and production yields of these chemically complex compounds, genetic manipulation of the producer strains can be performed. Heterologous expression in amenable hosts can be useful to exploit and to explore the genetic potential of actinomycetes and not cultivable but interesting bacteria. Artificial chromosomes that can be stably integrated into the Streptomyces genome were constructed and demonstrated to be effective for transferring entire biosynthetic gene clusters from intractable actinomycetes into more suitable hosts. In this paper, the construction of several shuttle Escherichia coli-Streptomyces artificial chromosomes is discussed together with old and new strategies applied to improve heterologous production of secondary metabolites.

  7. Antimicrobial biosynthetic potential and genetic diversity of endophytic actinomycetes associated with medicinal plants.

    PubMed

    Gohain, Anwesha; Gogoi, Animesh; Debnath, Rajal; Yadav, Archana; Singh, Bhim P; Gupta, Vijai K; Sharma, Rajeev; Saikia, Ratul

    2015-10-01

    Endophytic actinomycetes are one of the primary groups that share symbiotic relationships with medicinal plants and are key reservoir of biologically active compounds. In this study, six selective medicinal plants were targeted for the first time for endophytic actinomycetes isolation from Gibbon Wild Life Sanctuary, Assam, India, during winter and summer and 76 isolates were obtained. The isolates were found to be prevalent in roots followed by stem and leaves. 16S rRNA gene sequence analysis revealed 16 genera, including rare genera, Verrucosispora, Isoptericola and Kytococcus, which have never been previously reported as endophytic. The genus Streptomyces (66%) was dominant in both seasons. Shannon's diversity index showed that Azadirachta indica (1.49), Rauwolfia serpentina (1.43) and Emblica officinalis (1.24) were relatively good habitat for endophytic actinomycetes. Antimicrobial strains showed prevalence of polyketide synthase (PKS) type-II (85%) followed by PKS type-I (14%) encoded in the genomes. Expression studies showed 12-fold upregulation of PKSII gene in seventh day of incubation for Streptomyces antibioticus (EAAG90). Our results emphasize that the actinomycetes assemblages within plant tissue exhibited biosynthetic systems encoding for important biologically active compounds.

  8. Novel bioactive compounds from actinomycetes.

    PubMed

    Sanglier, J J; Wellington, E M; Behal, V; Fiedler, H P; Ellouz Ghorbel, R; Finance, C; Hacene, M; Kamoun, A; Kelly, C; Mercer, D K

    1993-10-01

    Actinomycetes form an enormous reservoir of secondary metabolites and enzymes. The potential for exploiting rare actinomycetes is highlighted by the discovery of novel compounds from strains of Spirillospora and Nocardioides. Novel compounds of well known classes of antibiotics, such as polyenes, continue to be discovered. For compounds containing a chromophore, the analysis by high-performance liquid chromatography coupled with a diode-array detector enables the elimination of producers of known compounds and facilitates the discovery of novel compounds or derivatives. The complexity of the regulatory mechanisms is illustrated by glutamine synthetase. The characterization of thermostable amylolytic, lignolytic, peroxidase and neuramidase activities, and the isolation of novel cellulolytic actinomycetes clearly demonstrate the potential of Actinomycetes as producers of enzymes.

  9. Diversity and bioprospecting of culturable actinomycetes from marine sediment of the Yellow Sea, China.

    PubMed

    Xiong, Zhi-Qiang; Liu, Qiao-Xia; Pan, Zhao-Long; Zhao, Na; Feng, Zhi-Xiang; Wang, Yong

    2015-03-01

    Marine actinomycetes are a potential source of a wide variety of bioactive natural products. In this work, seven pretreatments, three selective isolation media, and five artificial seawater concentrations were used to isolate actinomycetes from the sediments collected from Yellow Sea, China. Statistical analysis showed that only the isolation medium strongly affected the total and bioactive numbers of actinomycete isolates. A total of 613 actinobacterial strains were isolated and screened for antimicrobial activities; 154 isolates showed activity against at least one of nine test drug-resistant microorganisms. Eighty-nine representatives with strong antimicrobial activity were identified phylogenetically based on 16S rRNA gene sequencing, which were assigned to five different actinomycete genera Streptomyces, Kocuria, Saccharomonospora, Micromonospora, and Nocardiopsis. Using PCR-based screening for six biosynthetic genes of secondary metabolites, all 45 isolates with acute activity have at least one biosynthetic gene, 28.8 % of which possess more than three biosynthetic genes. As a case, strain SMA-1 was selected for antimicrobial natural product discovery. Three diketopiperazine dimers including a new compound iso-naseseazine B (1) and two known compounds naseseazine B (2) and aspergilazine A (3) were isolated by bioassay-guided separation. These results suggested that actinomycetes from marine sediments are a potential resource of novel secondary metabolites and drugs.

  10. Comparison of the Cell-Wall Composition of Morphologically Distinct Actinomycetes

    PubMed Central

    Yamaguchi, Tatsuro

    1965-01-01

    Yamaguchi, Tatsuro (The University of Tokyo, Tokyo, Japan). Comparison of the cell-wall composition of morphologically distinct actinomycetes. J. Bacteriol. 89:444–453. 1965.—Cell-wall composition of various morphologically distinct actinomycetes was studied to determine the relationship, if any, between cell-wall composition and morphological criteria in actinomycete taxonomy. The methods used were similar to those of Cummins and Harris. At least five types of cell-wall composition were obtained; however, these were not always correlated with groupings by the conventional classification system. For instance, the sporangium-forming actinomycetes, Actinoplanaceae, had three types of cell-wall composition; the composition of cell walls of Promicromonospora, Micromonospora, and Microbispora was the same as, or similar to, that of Actinomyces, Actinoplanes, and Streptosporangium, respectively; Chainia, Actinopycnidium, Actinosporangium, and Microellobosporia had the same cell-wall composition as Streptomyces, whereas that of Streptoverticillium was slightly different. Possible implications of cell-wall composition and morphological differentiation of hyphae for the taxonomy and phylogeny of actinomycetes are also discussed. PMID:14255713

  11. The putative elaiophylin biosynthetic gene cluster in Streptomyces sp. DSM4137 is adjacent to genes encoding adenosylcobalamin-dependent methylmalonyl CoA mutase and to genes for synthesis of cobalamin.

    PubMed

    Haydock, Stephen F; Mironenko, Tatiana; Ghoorahoo, Haroun I; Leadlay, Peter F

    2004-09-30

    A type I PKS gene probe obtained from RAPB of the rapamycin producer Streptomyces hygroscopicus, strongly hybridised to 92 out of 1120 cosmids from a genomic library of the elaiophylin-producing strain Streptomyces sp. DSM4137. Partial cosmid sequencing suggested the presence of 10 separate sequences encoding type I PKS genes. One entire DNA sequence was obtained and found exactly to match the gene organisation expected for the biosynthesis of the unusual macrodiolide polyketide elaiophylin. The putative elaiophylin gene cluster contains five large open-reading frames encoding typical modular polyketide synthases, which together catalyse the synthesis of the octaketide monomer of elaiophylin. Other genes were identified that would be required for provision of the ethylmalonate extender unit, for the synthesis and attachment of 2-deoxy-L-fucose and in regulation, or in export of the product. Immediately adjacent to the putative elaiophylin biosynthetic gene cluster is a 30-kbp region containing the gene for adenosylcobalamin-dependent methylmalonyl CoA mutase and also genes involved in the biosynthesis of the cobalamin cofactor. Analysis of the latter gene set confirms the view that cbiD of the anaerobic pathway and cobF in the aerobic pathway catalyse the same methylation of precorrin-5. The proximity of these genes to the putative elaiophylin gene cluster can best be rationalised if in this organism succinyl-CoA is a significant source of the methylmalonate units for complex polyketide biosynthesis.

  12. Degradation of carbonyl sulfide by Actinomycetes and detection of clade D of β-class carbonic anhydrase.

    PubMed

    Ogawa, Takahiro; Kato, Hiromi; Higashide, Mitsuru; Nishimiya, Mami; Katayama, Yoko

    2016-09-25

    Carbonyl sulfide (COS) is an atmospheric trace gas and one of the sources of stratospheric aerosol contributing to climate change. Although one of the major sinks of COS is soil, the distribution of COS degradation ability among bacteria remains unclear. Seventeen out of 20 named bacteria belonging to Actinomycetales had COS degradation activity at mole fractions of 30 parts per million by volume (ppmv) COS. Dietzia maris NBRC 15801(T) and Mycobacterium sp. THI405 had the activity comparable to a chemolithoautotroph Thiobacillus thioparus THI115 that degrade COS by COS hydrolase for energy production. Among 12 bacteria manifesting rapid degradation at 30 ppmv COS, Dietzia maris NBRC 15801(T) and Streptomyces ambofaciens NBRC 12836(T) degraded ambient COS (∼500 parts per trillion by volume). Geodermatophilus obscurus NBRC 13315(T) and Amycolatopsis orientalis NBRC 12806(T) increased COS concentrations. Moreover, six of eight COS degrading bacteria isolated from soils had partial nucleotide sequences similar to that of the gene encoding clade D of β-class carbonic anhydrase, which included COS hydrolase. These results indicate the potential importance of Actinomycetes in the role of soils as sinks of atmospheric COS.

  13. Characterization of a gamma-butyrolactone synthetase gene homologue (stcA) involved in bafilomycin production and aerial mycelium formation in Streptomyces sp. SBI034.

    PubMed

    Intra, Bungonsiri; Euanorasetr, Jirayut; Nihira, Takuya; Panbangred, Watanalai

    2016-03-01

    Streptomyces SBI034 produces several bafilomycin derivatives. Its afsA homologue (stcA) and putative γ-butyrolactone receptor gene (stcB) were cloned. Construction of a stcA disruptant (stcA gene knockout) resulted in complete abolishment of all bafilomycin production. Electron microscopic analysis showed a defect of aerial mycelium formation and sporulation in the stcA disruptant. Restoration of all phenotypic defects and bafilomycin production was observed in a stcA complemented strain. Addition of exogenous γ-butyrolactone (GBL) extracted from the culture broth of the wild-type strain could stimulate the aerial mycelium and spore formation of the stcA disruptant. These results suggest that stcA plays a role in GBL-mediated regulation of bafilomycin biosynthesis and morphological development in Streptomyces strain SBI034.

  14. Taxonomic study of a salt tolerant Streptomyces sp. strain C-2012 and the effect of salt and ectoine on lon expression level.

    PubMed

    Sadeghi, Akram; Soltani, Bahram M; Jouzani, Gholamreza Salehi; Karimi, Ebrahim; Nekouei, Mojtaba Khayam; Sadeghizadeh, Majid

    2014-01-01

    Streptomyces strain C-2012 is a salt tolerant biocontrol PGPR that has been isolated from Iranian soil. The main aim of current study was finding strain C-2012 taxonomic position and to find the genes which are potentially involved in salt tolerance phenotype. Strain C-2012 chemotaxonomic, morphological and molecular characteristics indicate that this strain is a member of the genus Streptomyces. Phylogenetic analyses based on an almost complete 16S rRNA gene sequence revealed that this strain is closely related to Streptomyces rimosus JCM 4667(T). Also, DNA-DNA hybridization test estimated 74% relatedness between two strains and confirmed that C-2012 is a strain of S. rimosus. In order to find novel genes that are differentially expressed in response to the salt treatment, cDNA-AFLP was carried out. One of the selected expressed sequence tags (TDF-1) was found to be homologous to lon gene which produces a bacterial ATP-dependent proteases (proteases LA). Lon gene expression was induced following 450 mM salt (NaCl) treatment and its expression level was further (5.2-fold) increased in response to salt when ectoine was added to the medium. These results suggest that two protein protection systems including ectoine and ATP-dependent proteases synergistically are related. NaCl stress also caused an enhancement in the activity of extracellular protease.

  15. Enhancement of ε-poly-L-lysine production coupled with precursor L-lysine feeding in glucose-glycerol co-fermentation by Streptomyces sp. M-Z18.

    PubMed

    Chen, Xu-Sheng; Ren, Xi-Dong; Zeng, Xin; Zhao, Fu-Lin; Tang, Lei; Zhang, Hong-Jian; Zhang, Jian-Hua; Mao, Zhong-Gui

    2013-12-01

    ε-Poly-L-lysine (ε-PL), one of the only two homo-poly amino acids known in nature, is used as a preservative. In this study, strategies of feeding precursor L-lysine into 5 L laboratory scale fermenters, including optimization of L-lysine concentration and time, was investigated to optimize the production of ε-PL by Streptomyces sp. M-Z18. The optimized strategy was then used in ε-PL fed-batch fermentation in which glucose and glycerol served as mixed carbon sources. In this way, a novel ε-PL production strategy involving precursor L-lysine coupled with glucose-glycerol co-fermentation was developed. Under optimal conditions, ε-PL production reached 37.6 g/l, which was 6.2 % greater than in a previous study in which glucose and glycerol co-fermentation was performed without added L-lysine (35.14 g/l). To the best of our knowledge, this is the first report of the enhancement of ε-PL production through L-lysine feeding to evaluate the use of fermenters. Meanwhile, the role of L-lysine in the promotion of ε-PL production, participating ε-PL synthesis as a whole, was first determined using the L-[U-(13)C] lysine labeling method. It has been suggested that the bottleneck of ε-PL synthesis in Streptomyces sp. M-Z18 is in the biosynthesis of precursor L-lysine. The information obtained in the present work may facilitate strain improvement and efficient large-scale ε-PL production.

  16. [Selective Isolation of Rare Actinomycetes from Soil].

    PubMed

    Sineva, O N; Terekhova, L P

    2015-01-01

    Many diverse methods for selective isolation of actinomycetes are used in discovery of organisms producing biologically active substances, as well as in ecological studies. Methods for isolation of rare actinomycetes from soil are reviewed.

  17. Presence, molecular characteristics and geosmin producing ability of actinomycetes isolated from South Korean terrestrial and aquatic environments.

    PubMed

    Lee, Gyu-Cheol; Kim, Yun S; Kim, Min-Jeong; Oh, Sung-Ae; Choi, Ilhwan; Choi, Jaewon; Park, Jong-Geun; Chong, Chom-Kyu; Kim, Yong-Yeon; Lee, Kyeunghee; Lee, Chan Hee

    2011-01-01

    The unpleasant odor of drinking water is one of the major problems in many water utilities in the world. Actinomycetes have long been associated with odorous compounds. Considering the paucity of research on Actinomycetes producing odorous compounds in South Korea, presence of Actinomycetes, their molecular characteristics and ability to produce odorous compounds were investigated in this study. Findings confirmed the presence of Actinomycetes in surface soil, sediment, and water samples from four sites: two artificial lakes [Paldang and Cheongpyeong (CP)], and two streams [Gyeongan (GA) and Yangpyeong]. Surface soil and sediment from CP area had the greatest concentration of Actinomycetes (8.2 x 10(7) and 6.8 x 10(6) colony forming units (CFUs)/gram, dry weight, respectively). When water samples are considered, samples from GA had the highest concentration (1.9 x 10(2) CFU/mL). 16S rRNA sequencing and molecular phylogenetic analysis showed that Streptomyces was the dominant genus (64.1%). In addition, the isolated Actinomycetes synthesized 5.4 ng/L geosmin as demonstrated by thermal desorption unit-gas chromatograph/mass spectrometry analysis.

  18. Actinomycetes from solitary wasp mud nest and swallow bird mud nest: isolation and screening for their antibacterial activity.

    PubMed

    Kumar, Vijay; Bharti, Alpana; Gupta, Vivek Kumar; Gusain, Omprakash; Bisht, Gajraj Singh

    2012-03-01

    The aim of this study was to isolate and screen actinomycetes from solitary wasp and swallow bird mud nests for antimicrobial activity. The actinomycetes were isolated from soil of nests of solitary wasp and swallow bird, and identified on the basis of morphological characteristics and molecular biological methods. A total of 109 actinomycetal isolates were obtained from 12 soil samples (6 from each habitat) using two media. The highest number of actinomycetes were recovered on Humic acid vitamin agar media (65.13%, n = 71) as compared to actinomycetes isolation agar media (34.86%, n = 38). The antimicrobial activity of actinomycetes isolates was determined using the agar plug method. Among 109 isolates, 51 isolates (46.78%) showed antibacterial activity by agar plug assay. The morphological and molecular characteristics confirmed that the most of active isolates in both sample belonged to the genus Streptomyces, the other potential genera like Streptosporangium, Actinomadura, Saccharopolyspora, Thermoactinomycetes and Nocardia were also recovered, but in a low frequency. The isolates designated as 8(1), BN-6, MN 2(6), MN 2(7) and MN 9(V) showed most promising activity against various drug resistant bacterial pathogens. It seems that the promising isolates from these unusual/unexplored habitats may prove to be an important step in development of drug for treating multi-drug resistant bacterial pathogens.

  19. Unique actinomycetes from marine caves and coral reef sediments provide novel PKS and NRPS biosynthetic gene clusters.

    PubMed

    Hodges, Tyler W; Slattery, Marc; Olson, Julie B

    2012-06-01

    In the ever-expanding search for novel bioactive molecules and enzymes, marine actinomycetes have proven to be a productive source. While open reef sediment and sponge-associated actinomycetes have been extensively examined, their marine cave counterparts remain unevaluated. Anchialine cave systems in the Bahamas offered an ideal setting to evaluate the occurrence and variation within sediment-associated actinomycete communities. While in close geographical proximity to open reef environments, these systems provide a specialized environmental niche devoid of light and direct exposure to nutrient input. In the present study, selective isolation techniques and molecular methods were used to test the hypothesis that variable distribution of actinomycetes and secondary metabolite gene clusters occur between open reef and marine cave systems. The results indicated that differences exist within the culturable sediment-associated actinomycete communities between marine caves and open reef systems, with members of the genus Streptomyces dominating cultures from open reef sediments and a more diverse suite of actinomycetes isolated from marine cave sediment samples. Within the cave isolates, members of the proposed genus Solwaraspora were the most represented. Based on PKS- and NRPS-gene-targeted PCR amplification and sequencing, geographic variation in the occurrence of these biosynthetic pathways was also observed. These findings indicate that marine cave systems are a lucrative source in the search for novel secondary metabolite producers with biotechnological applications and that environmental and geographic factors likely affect the occurrence of these biosynthetic pathways.

  20. Melanogenic actinomycetes from rhizosphere soil-antagonistic activity against Xanthomonas oryzae and plant-growth-promoting traits.

    PubMed

    Muangham, Supattra; Pathom-Aree, Wasu; Duangmal, Kannika

    2015-02-01

    A total of 210 melanogenic actinomycetes were isolated from 75 rhizospheric soils using ISP6 and ISP7 agar supplemented with antifungal and antibacterial agents. Their morphological characteristics and the presence of ll-diaminopimelic acid in whole-cell hydrolyzates revealed that all isolates belonged to the genus Streptomyces. Their ability to inhibit the growth of 2 pathogenic rice bacteria, Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola, was observed using the agar overlay method. The results indicated that 61.9% of the isolates could inhibit at least one of the tested rice pathogens. Among these, isolate TY68-3 showed the highest antibacterial activity and siderophore production. The 16S rRNA gene sequence analysis of 46 representative isolates revealed that isolates with high similarity to Streptomyces bungoensis were frequently found. The present study indicated the potential of melanogenic actinomycetes for use as biocontrol agents against X. oryzae as well as their diversity in rhizospheric soils.

  1. New benzoxazine secondary metabolites from an arctic actinomycete.

    PubMed

    Moon, Kyuho; Ahn, Chan-Hong; Shin, Yoonho; Won, Tae Hyung; Ko, Keebeom; Lee, Sang Kook; Oh, Ki-Bong; Shin, Jongheon; Nam, Seung-Il; Oh, Dong-Chan

    2014-04-30

    Two new secondary metabolites, arcticoside (1) and C-1027 chromophore-V (2), were isolated along with C-1027 chromophore-III and fijiolides A and B (3-5) from a culture of an Arctic marine actinomycete Streptomyces strain. The chemical structures of 1 and 2 were elucidated through NMR, mass, UV, and IR spectroscopy. The hexose moieties in 1 were determined to be d-glucose from a combination of acid hydrolysis, derivatization, and gas chromatographic analyses. Arcticoside (1) and C-1027 chromophore-V (2), which have a benzoxazine ring, inhibited Candida albicans isocitrate lyase. Chromophore-V (2) exhibited significant cytotoxicity against breast carcinoma MDA-MB231 cells and colorectal carcinoma cells (line HCT-116), with IC₅₀ values of 0.9 and 2.7 μM, respectively.

  2. Discovery of novel metabolites from marine actinomycetes.

    PubMed

    Lam, Kin S

    2006-06-01

    Recent findings from culture-dependent and culture-independent methods have demonstrated that indigenous marine actinomycetes exist in the oceans and are widely distributed in different marine ecosystems. There is tremendous diversity and novelty among the marine actinomycetes present in marine environments. Progress has been made to isolate novel actinomycetes from samples collected at different marine environments and habitats. These marine actinomycetes produce different types of new secondary metabolites. Many of these metabolites possess biological activities and have the potential to be developed as therapeutic agents. Marine actinomycetes are a prolific but underexploited source for the discovery of novel secondary metabolites.

  3. Taxonomic and ecological studies of actinomycetes from Vietnam: isolation and genus-level diversity.

    PubMed

    Hop, Duong Van; Sakiyama, Yayoi; Binh, Chu Thi Thanh; Otoguro, Misa; Hang, Dinh Thuy; Miyadoh, Shinji; Luong, Dao Thi; Ando, Katsuhiko

    2011-09-01

    Actinomycetes were isolated from 109 soil and 93 leaf-litter samples collected at five sites in Vietnam between 2005 and 2008 using the rehydration-centrifugation (RC) method, sodium dodecyl sulfate-yeast extract dilution method, dry-heating method and oil-separation method in conjunction with humic acid-vitamin agar as an isolation medium. A total of 1882 strains were identified as Vietnamese (VN)-actinomycetes including 1080 (57%) streptomycetes (the genus Streptomyces isolates) and 802 (43%) non-streptomycetes. The 16S ribosomal RNA gene sequences of the VN-actinomycetes were analyzed using BLAST searches. The results showed that these isolates belonged to 53 genera distributed among 21 families. Approximately 90% of these strains were members of three families: Streptomycetaceae (1087 strains, 58%); Micromonosporaceae (516 strains, 27%); and Streptosporangiaceae (89 strains, 5%). Motile actinomycetes of the genera Actinoplanes, Kineosporia and Cryptosporangium, which have quite common morphological characteristics, were frequently isolated from leaf-litter samples using the RC method. It is possible that these three genera acquired common properties during a process of convergent evolution. By contrast, strains belonging to the suborder Streptosporangineae were exclusively isolated from soils. A comparison of the sampling sites revealed no significant difference in taxonomic diversity between these sites. Among the non-streptomycetes, 156 strains (19%) were considered as new taxa distributed into 21 genera belonging to 12 families. Interestingly, the isolation of actinomycetes from leaf-litter samples using the RC method proved to be the most efficient way to isolate new actinomycetes in Vietnam, especially the Micromonosporaceae species.

  4. [Progress in developing and applying Streptomyces chassis - A review].

    PubMed

    Xiao, Liping; Deng, Zixin; Liu, Tiangang

    2016-03-04

    Natural products and their derivatives play an important role in modern healthcare. Their diversity in bioactivity and chemical structure inspires scientists to discover new drug entities for clinical use. However, chemical synthesis of natural compounds has insurmountable difficulties in technology and cost. Also, many original-producing bacteria have disadvantages of needing harsh cultivation conditions, having low productivity and other shortcomings. In addition, some gene clusters responsible for secondary metabolite biosynthesis are silence in the original strains. Therefore, it is of great significance to exploit strategy for the heterologous expression of natural products guided by synthetic biology. Recently, researchers pay more attention on using actinomycetes that are the main source of many secondary metabolites, such as antibiotics, anticancer agents, and immunosuppressive drugs. Especially, with huge development of genome sequencing, abundant resources of natural product biosynthesis in Streptomyces have been discovered, which highlight the special advantages on developing Streptomyces as the heterologous expression chassis cells. This review begins with the significance of the development of Streptomyces chassis, focusing on the strategies and the status in developing Streptomyces chassis cells, followed by examples to illustrate the practical applications of a variety of Streptomyces chassis.

  5. Red Soils Harbor Diverse Culturable Actinomycetes That Are Promising Sources of Novel Secondary Metabolites

    PubMed Central

    Guo, Xiaoxuan; Liu, Ning; Li, Xiaomin; Ding, Yun; Shang, Fei; Gao, Yongsheng; Ruan, Jisheng

    2015-01-01

    Red soils, which are widely distributed in tropical and subtropical regions of southern China, are characterized by low organic carbon, high content of iron oxides, and acidity and, hence, are likely to be ideal habitats for acidophilic actinomycetes. However, the diversity and biosynthetic potential of actinomycetes in such habitats are underexplored. Here, a total of 600 actinomycete strains were isolated from red soils collected in Jiangxi Province in southeast China. 16S rRNA gene sequence analysis revealed a high diversity of the isolates, which were distributed into 26 genera, 10 families, and 7 orders within the class Actinobacteria; these taxa contained at least 49 phylotypes that are likely to represent new species within 15 genera. The isolates showed good physiological potentials for biosynthesis and biocontrol. Chemical screening of 107 semirandomly selected isolates spanning 20 genera revealed the presence of at least 193 secondary metabolites from 52 isolates, of which 125 compounds from 39 isolates of 12 genera were putatively novel. Macrolides, polyethers, diketopiperazines, and siderophores accounted for most of the known compounds. The structures of six novel compounds were elucidated, two of which had a unique skeleton and represented characteristic secondary metabolites of a putative novel Streptomyces phylotype. These results demonstrate that red soils are rich reservoirs for diverse culturable actinomycetes, notably members of the families Streptomycetaceae, Pseudonocardiaceae, and Streptosporangiaceae, with the capacity to synthesize novel bioactive compounds. PMID:25724963

  6. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges

    PubMed Central

    Cheng, Cheng; MacIntyre, Lynsey; Abdelmohsen, Usama Ramadan; Horn, Hannes; Polymenakou, Paraskevi N.; Edrada-Ebel, RuAngelie; Hentschel, Ute

    2015-01-01

    Marine sponge–associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes. PMID:26407167

  7. Population densities and genetic diversity of actinomycetes associated to the rhizosphere of Theobroma cacao

    PubMed Central

    Barreto, Tâmara R.; da Silva, Augusto C.M.; Soares, Ana Cristina F.; de Souza, Jorge T.

    2008-01-01

    In spite of the acknowledged importance of growth-promoting bacteria, only a reduced number of studies were conducted with these microorganisms on Theobroma cacao. The objectives of this work were to study the population densities and genetic diversity of actinomycetes associated with the rhizosphere of cacao as a first step in their application in plant growth promotion and biological control. The populations densities of actinomycetes in soil and cacao roots were similar, with mean values of 1,0 x 106 CFU/g and 9,6 x 105 CFU/g, respectively. All isolates selected and used in this study were identified through sequencing analyses of a fragment of the rpoB gene that encodes the β-subunit of the RNA polymerase as species of the genus Streptomyces. In vitro cellulolytic, xilanolytic and chitinolytic activity, indolacetic acid production and phosphate solubilization activities were observed in most of the isolates tested. The data obtained in this study demonstrate that actinomycetes account for a higher percentage of the total population of culturable bacteria in soil than on cacao roots. Additionally, actinomycetes from the cacao rhizosphere are genetically diverse and have potential applications as agents of growth promotion. PMID:24031247

  8. Eliciting antibiotics active against the ESKAPE pathogens in a collection of actinomycetes isolated from mountain soils.

    PubMed

    Zhu, Hua; Swierstra, Jasper; Wu, Changsheng; Girard, Geneviève; Choi, Young Hae; van Wamel, Willem; Sandiford, Stephanie K; van Wezel, Gilles P

    2014-08-01

    The rapid emergence of multidrug-resistant (MDR) bacterial pathogens poses a major threat for human health. In recent years, genome sequencing has unveiled many poorly expressed antibiotic clusters in actinomycetes. Here, we report a well-defined ecological collection of >800 actinomycetes obtained from sites in the Himalaya and Qinling mountains, and we used these in a concept study to see how efficiently antibiotics can be elicited against MDR pathogens isolated recently from the clinic. Using 40 different growth conditions, 96 actinomycetes were identified - predominantly Streptomyces - that produced antibiotics with efficacy against the MDR clinical isolates referred to as ESKAPE pathogens: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and/or Enterobacter cloacae. Antimicrobial activities that fluctuated strongly with growth conditions were correlated with specific compounds, including borrelidin, resistomycin, carbomethoxy-phenazine, and 6,7,8- and 5,6,8-trimethoxy-3-methylisocoumarin, of which the latter was not described previously. Our work provided insights into the potential of actinomycetes as producers of drugs with efficacy against clinical isolates that have emerged recently and also underlined the importance of targeting a specific pathogen.

  9. Red soils harbor diverse culturable actinomycetes that are promising sources of novel secondary metabolites.

    PubMed

    Guo, Xiaoxuan; Liu, Ning; Li, Xiaomin; Ding, Yun; Shang, Fei; Gao, Yongsheng; Ruan, Jisheng; Huang, Ying

    2015-05-01

    Red soils, which are widely distributed in tropical and subtropical regions of southern China, are characterized by low organic carbon, high content of iron oxides, and acidity and, hence, are likely to be ideal habitats for acidophilic actinomycetes. However, the diversity and biosynthetic potential of actinomycetes in such habitats are underexplored. Here, a total of 600 actinomycete strains were isolated from red soils collected in Jiangxi Province in southeast China. 16S rRNA gene sequence analysis revealed a high diversity of the isolates, which were distributed into 26 genera, 10 families, and 7 orders within the class Actinobacteria; these taxa contained at least 49 phylotypes that are likely to represent new species within 15 genera. The isolates showed good physiological potentials for biosynthesis and biocontrol. Chemical screening of 107 semirandomly selected isolates spanning 20 genera revealed the presence of at least 193 secondary metabolites from 52 isolates, of which 125 compounds from 39 isolates of 12 genera were putatively novel. Macrolides, polyethers, diketopiperazines, and siderophores accounted for most of the known compounds. The structures of six novel compounds were elucidated, two of which had a unique skeleton and represented characteristic secondary metabolites of a putative novel Streptomyces phylotype. These results demonstrate that red soils are rich reservoirs for diverse culturable actinomycetes, notably members of the families Streptomycetaceae, Pseudonocardiaceae, and Streptosporangiaceae, with the capacity to synthesize novel bioactive compounds.

  10. Detection of polyketide synthase and nonribosomal peptide synthetase biosynthetic genes from antimicrobial coral-associated actinomycetes.

    PubMed

    Li, Jie; Dong, Jun-De; Yang, Jian; Luo, Xiong-Ming; Zhang, Si

    2014-10-01

    The diversity and properties of actinobacteria, predominant residents in coral holobionts, have been rarely documented. In this study, we aimed to explore the species diversity, antimicrobial activities and biosynthetic potential of culturable actinomycetes within the tissues of the scleractinian corals Porites lutea, Galaxea fascicularis and Acropora millepora from the South China Sea. A total of 70 strains representing 13 families and 15 genera of actinobacteria were isolated. The antimicrobial activity and biosynthetic potential of fifteen representative filamentous actinomycetes were estimated. Crude fermentation extracts of 6 strains exhibited comparable or greater activities against Vibrio alginolyticus than ciprofloxacin. Seven of the 15 actinomycetes strains possess type I polyketide synthases (PKS-I) and/or nonribosomal peptide synthetases (NRPS) genes. Nine tested strains possess type II polyketide synthases (PKS-II). Phylogenetic analysis based on 16S rRNA gene sequences indicated that these PKS and NRPS gene screening positive strains belong to genera Nocardiopsis, Pseudonocardia, Streptomyces, Micromonospora, Amycolatopsis and Prauserella. One PKS-I and four NRPS fragments showed <70% similarity to their closest relatives, which suggested the novelty of these genes. This study helps uncover the genetic capacity of stony coral-associated actinomycetes to produce bioactive molecules.

  11. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges.

    PubMed

    Cheng, Cheng; MacIntyre, Lynsey; Abdelmohsen, Usama Ramadan; Horn, Hannes; Polymenakou, Paraskevi N; Edrada-Ebel, RuAngelie; Hentschel, Ute

    2015-01-01

    Marine sponge-associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes.

  12. Discaria trinervis - Frankia symbiosis promotion by saprophytic actinomycetes.

    PubMed

    Solans, Mariana

    2007-06-01

    The influence of saprophytic actinomycetes strains on the Discaria trinervis - Frankia actinorhizal symbiosis was investigated. Three strains out of 122 isolated from the rhizosphere and rhizoplane of D. trinervis with multiple enzymatic activities, were selected for plant growth experiments: Streptomyces (BCRU-MM40), Actinoplanes (BCRU-ME3) and Micromonospora (BCRU-MM18). Inoculated seedlings of Discaria trinervis were grown in glass tubes with vermiculite-sand for 12 weeks. They were inoculated either with a single saprophytic strain or a combination of one or two of them together with the symbiotic N(2) fixing strain Frankia BCU110501. The saprophytic strains were applied in two experimental series, i.e. mycelium + supernatant simultaneously or mycelium and supernatant (growth medium free of cells) separately. Micromonospora strain MM18 showed a direct promotion effect on shoot growth, when plants were inoculated with mycelium and supernatant together. Streptomyces strain MM40 and Actinoplanes strain ME3 promoted the actinorhizal symbiosis with Frankia and consequently the development of plant shoots, when supernatant was involved as inoculum. It is supposed, that the strains MM18, MM40 and ME3 produce bioactive metabolites, which are released into the culture medium. The saprophytic strains studied could be considered as "promoting or helper rhizoactinomycetes" of the actinorhizal plant D. trinervis.

  13. Rapid identification of filamentous actinomycetes to the genus level using genus-specific 16S rRNA gene restriction fragment patterns.

    PubMed

    Cook, Andrew E; Meyers, Paul R

    2003-11-01

    A rapid method for identifying filamentous actinomycete genera was developed based on 16S rRNA gene restriction fragment patterns. The patterns were generated by using specific restriction endonucleases to perform in silico digestions on the 16S rRNA gene sequences of all validly published filamentous actinomycete species. The method was applied to identifying actinomycete isolates from soil. Amplified 16S rDNA of soil actinomycetes was restricted with selected endonucleases and electrophoresed on agarose gels. The restriction fragment patterns of the unknown isolates were easily compared to the established patterns. Significantly, the genus Streptomyces could be differentiated from all other actinomycete genera by using only four restriction endonucleases, Sau3AI, AsnI, KpnI and SphI. This could be achieved in a time period of as little as a week, following PCR-template DNA isolation by a simple method. The identification method allowed unknown, non-Streptomyces soil isolates to be identified to a genus or small subgroup of genera. The genera in these subgroups could, in some cases, be distinguished by virtue of colony-morphology differences.

  14. Isolation and characterization of cyclo-(tryptophanyl-prolyl) and chloramphenicol from Streptomyces sp. SUK 25 with antimethicillin-resistant Staphylococcus aureus activity

    PubMed Central

    Alshaibani, Muhanna M; Jalil, Juriyati; Sidik, Nik M; Edrada-Ebel, Ruangelie; Zin, Noraziah M

    2016-01-01

    Background Zingiber spectabile, commonly known as Beehive Ginger, is used as an ethnobotanical plant in many countries as an appetizer or to treat stomachache, toothache, muscle sprain, and as a cure for swelling, sores and cuts. This is the first report of isolation of Streptomyces strain from the root of this plant. Strain Universiti Kebangsaan 25 (SUK 25) has a very high activity to produce secondary metabolites against methicillin-resistant Staphylococcus aureus (MRSA), which is associated with high morbidity and mortality rates due to acquired multidrug resistance genes and causes medication failure in some clinical cases worldwide. Phylogenetic analysis based on the 16S ribosomal RNA gene sequence exhibited that the most closely related strain was Streptomyces omiyaensis NBRC 13449T (99.0% similarity). Aim This study was conducted to carry out the extraction, identification, and biological evaluation of active metabolites isolated from SUK 25 against three MRSA strains, namely, MRSA ATCC 43300, MRSA ATCC 33591, and MRSA ATCC 49476. Materials and methods The production of secondary metabolites by this strain was optimized through Thronton’s media. Isolation, purification, and identification of the bioactive compounds were carried out using reversed-phase high-performance liquid chromatography, high-resolution mass spectrometry, Fourier transform infrared, and one-dimensional and two-dimensional nuclear magnetic resonance. Results During screening procedure, SUK 25 exhibited good antimicrobial potential against several strains of MRSA. The best biological activity was shown from fraction number VII and its subfractions F2 and F3 with minimum inhibitory concentration values at 16 µg/mL and 8 µg/mL, respectively. These two subfractions were identified as diketopiperazine cyclo-(tryptophanyl-prolyl) and chloramphenicol. Conclusion On the basis of obtained results, SUK 25 isolated from Z. spectabile can be regarded as a new valuable source to produce secondary

  15. Actinomycete integrative and conjugative pMEA-like elements of Amycolatopsis and Saccharopolyspora decoded.

    PubMed

    te Poele, Evelien M; Samborskyy, Markiyan; Oliynyk, Markiyan; Leadlay, Peter F; Bolhuis, Henk; Dijkhuizen, Lubbert

    2008-05-01

    Actinomycete integrative and conjugative elements (AICEs) are present in diverse genera of the actinomycetes, the most important bacterial producers of bioactive secondary metabolites. Comparison of pMEA100 of Amycolatopsis mediterranei, pMEA300 of Amycolatopsis methanolica and pSE211 of Saccharopolyspora erythraea, and other AICEs, revealed a highly conserved structural organisation, consisting of four functional modules (replication, excision/integration, regulation, and conjugative transfer). Features conserved in all elements, or specific for a single element, are discussed and analysed. This study also revealed two novel putative AICEs (named pSE222 and pSE102) in the Sac. erythraea genome, related to the previously described pSE211 and pSE101 elements. Interestingly, pSE102 encodes a putative aminoglycoside phosphotransferase which may confer antibiotic resistance to the host. Furthermore, two of the six pSAM2-like insertions in the Streptomyces coelicolor genome described by Bentley et al. [Bentley, S.D., Chater, K.F., Cerdeno-Tarraga, A.M., et al., 2002. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417, 141-147] could be functional AICEs. Homologues of various AICE proteins were found in other actinomycetes, in Frankia species and in the obligate marine genus Salinispora and may be part of novel AICEs as well. The data presented provide a better understanding of the origin and evolution of these elements, and their functional properties. Several AICEs are able to mobilise chromosomal markers, suggesting that they play an important role in horizontal gene transfer and spread of antibiotic resistance, but also in evolution of genome plasticity.

  16. Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost.

    PubMed

    Habbeche, Amina; Saoudi, Boudjema; Jaouadi, Bassem; Haberra, Soumaya; Kerouaz, Bilal; Boudelaa, Mokhtar; Badis, Abdelmalek; Ladjama, Ali

    2014-04-01

    An extracellular thermostable keratinase (KERAK-29) was purified and biochemically characterized from a thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 newly isolated from Algerian poultry compost. The isolate exhibited high keratinase production when grown in chicken feather meal media (24,000 U/ml). Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 29,233.10-Da. The data revealed that the 25 N-terminal residue sequence displayed by KERAK-29 was TQADPPSWGLNNIDRQTAFTKATSI, which showed high homology with those of Streptomyces proteases. This keratinase was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP), which suggests that it belongs to the serine protease family. Using keratin azure as a substrate, the optimum pH and temperature values for keratinase activity were pH 10 and 70°C, respectively. KERAK-29 was stable between 20 and 60°C and pH 3 and 10 for 5 and 120 h, respectively, and its thermoactivity and thermostability were enhanced in the presence of 5 mM Mn(2+). Its catalytic efficiency was higher than that of the KERAB keratinase from Streptomyces sp. strain AB1. KERAK-29 was also noted to show high keratinolytic activity and significant stability in the presence of detergents, which made it able to accomplish the entire feather-biodegradation process on its own. The ability of the A. keratinilytica strain Cpt29 to grow and produce substantial levels of keratinase using feather as a substrate could open new promising opportunities for the valorization of keratin-containing wastes and reduction of its impacts on the environment.

  17. Novel Actinomycete Isolated from Bulking Industrial Sludge

    PubMed Central

    White, Johanna M.; Labeda, David P.; Lechevalier, Mary P.; Owens, James R.; Jones, Daniel D.; Gauthier, Joseph J.

    1986-01-01

    A novel actinomycete was the predominant filamentous microorganism in bulking activated sludge in a bench-scale reactor treating coke plant wastewater. The bacterium was isolated and identified as an actinomycete that is biochemically and morphologically similar to Amycolatopsis orientalis; however, a lack of DNA homology excludes true relatedness. At present, the isolate (NRRL B 16216) cannot be assigned to the recognized taxa of actinomycetes. Images PMID:16347238

  18. Release of ferulic acid and feruloylated oligosaccharides from sugar beet pulp by Streptomyces tendae.

    PubMed

    Ferreira, P; Diez, N; Faulds, C B; Soliveri, J; Copa-Patiño, J L

    2007-05-01

    Given several promising industrial applications of ferulic acid, this study was designed to identify actinomycete strains able to release high levels of this acid from sugar beet pulp (SBP). Out of 47 strains tested, 37% were found to release free ferulic acid from the growth substrate. One strain, identified as Streptomyces tendae by 16S RNA gene sequencing, was capable of releasing 80% of the ferulic acid ester-linked to the pectin in SBP after 5 days of growth. These data suggest that some actinomycetes are able to release ferulic acid and feruloylated oligosaccharides from SBP. During growth on SBP, it seems that Streptomyces species solubilize and release feruloylated oligosaccharides by specific carbohydrase activities before de-esterification and release of free ferulic acid.

  19. Antimicrobial Activities of Some Actinomycetes Isolated from Different Rhizospheric Soils in Tunisia.

    PubMed

    Trabelsi, Ines; Oves, Daniel; Manteca, Angel; Genilloud, Olga; Altalhi, Abdullah; Nour, Mohamed

    2016-08-01

    Fifty four isolates of actinomycetes were collected from four different rhizospheric soils: 18 strains from palm tree bark and soil, 12 strains from an olive field soil, 9 strains from a coastal forest, and 15 strains from an agriculture soil situated in the Algerian-Tunisian border (Oum Tboul). Based on morphological and cultural characters, the isolates were classified as Streptomyces (42 strains), Micromonospora (4 strains), Pseudonocardia (1 strain), Actinomadura (1 strain), Nocardia (1 strain), and non-Streptomyces (5 strains). More than half of the isolates inhibited at least one tested pathogenic microorganisms in liquid culture. In addition, antimicrobial activities of some strains were tested on solid culture. Several bioactive compounds were identified by liquid chromatography joined with low-resolution mass spectroscopy (LC/MS) and analysed by MEDINA's database and by the dictionary of natural products Chapman & Hall. An interesting chlorinated compound with the molecular formula C20H37ClN2O4, produced by three different strains (SF1, SF2, and SF5), was subject of an attempted purification. However, it was demonstrated using confocal microscopy and LC/MS high resolution that this compound is produced only on solid culture. These three potential antimicrobial isolates showed high similarity with Streptomyces thinghirensis and Streptomyces lienomycini, in terms of morphological characteristics and 16S rRNA gene sequences (bootstrap 97 %). All these findings prove the high antimicrobial diversity of the studied soils. The potential of the selected and other relatively unexplored extreme environments constitute a source of interesting actinomycete strains producing several biologically active secondary metabolites.

  20. Feeding of [5,5-2H(2)]-1-desoxy-D-xylulose and [4,4,6,6,6-2H(5)]-mevalolactone to a geosmin-producing Streptomyces sp. and Fossombronia pusilla.

    PubMed

    Spiteller, Dieter; Jux, Andreas; Piel, Jörn; Boland, Wilhelm

    2002-12-01

    The biosynthesis of the trisnor sesquiterpenoid geosmin (4,8a-dimethyl-octahydro-naphthalen-4a-ol) (1) was investigated by feeding labeled [5,5-2H(2)]-1-desoxy-D-xylulose (11), [4,4,6,6,6-(2)H(5)]-mevalolactone (7) and [2,2-2H(2)]-mevalolactone (9) to Streptomyces sp. JP95 and the liverwort Fossombronia pusilla. The micro-organism produced geosmin via the 1-desoxy-D-xylulose pathway, whereas the liverwort exclusively utilized mevalolactone for terpenoid biosynthesis. Analysis of the labeling pattern in the resulting isotopomers of geosmin (1) by mass spectroscopy (EI/MS) revealed that geosmin is synthesized in both organisms by cyclization of farnesyl diphosphate to a germacradiene-type intermediate 4. Further transformations en route to geosmin (1) involve an oxidative dealkylation of an i-propyl substituent, 1,2-reduction of a resulting conjugated diene, and bicyclization of a germacatriene intermediate 13. The transformations largely resemble the biosynthesis of dehydrogeosmin (2) in cactus flowers but differ with respect to the regioselectivity of the side chain dealkylation and 1,2-reduction

  1. Evaluation of Streptomyces strains isolated from herbal vermicompost for their plant growth-promotion traits in rice.

    PubMed

    Gopalakrishnan, Subramaniam; Vadlamudi, Srinivas; Bandikinda, Prakash; Sathya, Arumugam; Vijayabharathi, Rajendran; Rupela, Om; Kudapa, Himabindu; Katta, Krishnamohan; Varshney, Rajeev Kumar

    2014-01-20

    Six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140, CAI-155 and KAI-180, isolated from six different herbal vermi-composts were characterized for in vitro plant growth-promoting (PGP) properties and further evaluated in the field for PGP activity in rice. Of the six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140 and CAI-155 produced siderophores; CAI-13, CAI-93, CAI-155 and KAI-180 produced chitinase; CAI-13, CAI-140, CAI-155 and KAI-180 produced lipase; CAI-13, CAI-93, CAI-155 and KAI-180 produced protease; and CAI-13, CAI-85, CAI-140 and CAI-155 produced ß-1-3-glucanase whereas all the six actinomycetes produced cellulase, hydrocyanic acid and indole acetic acid (IAA). The actinomycetes were able to grow in NaCl concentrations of up to 8%, at pH values between 7 and 11, temperatures between 20 and 40 °C and compatible with fungicide bavistin at field application levels. In the rice field, the actinomycetes significantly enhanced tiller numbers, panicle numbers, filled grain numbers and weight, stover yield, grain yield, total dry matter, root length, volume and dry weight over the un-inoculated control. In the rhizosphere, the actinomycetes also significantly enhanced total nitrogen, available phosphorous, % organic carbon, microbial biomass carbon and nitrogen and dehydrogenase activity over the un-inoculated control. Sequences of 16S rDNA gene of the actinomycetes matched with different Streptomyces species in BLAST analysis. Of the six actinomycetes, CAI-85 and CAI-93 were found superior over other actinomycetes in terms of PGP properties, root development and crop productivity. qRT-PCR analysis on selected plant growth promoting genes of actinomycetes revealed the up-regulation of IAA genes only in CAI-85 and CAI-93.

  2. Challenges in the Heterologous Production of Antibiotics in Streptomyces.

    PubMed

    Bekiesch, Paulina; Basitta, Patrick; Apel, Alexander K

    2016-08-01

    The fast growing genome databases provide us with a large number of so far unknown secondary metabolite biosynthetic gene clusters. A key method to study these gene clusters is their heterologous expression in an engineered host strain. Gene clusters derived from actinomycetes are usually expressed in a Streptomyces host strain to identify and investigate the corresponding compounds. However, heterologous expression is often accompanied with some challenges affecting the production rates of secondary metabolites. The first step is therefore the selection of a suitable expression vector and host strain. Once production has been established, there are several possibilities to improve compound yields either by media screens, by overexpression of regulatory or transport genes or by introduction of constitutive or inducible promoters. A surely important, but hitherto little studied factor is also the regulation of a heterologously expressed gene cluster by its host strain. This review gives a short overview on the chances and challenges provided by heterologous production of secondary metabolites in Streptomyces.

  3. QM-HiFSA-Aided Structure Determination of Succinilenes A–D, New Triene Polyols from a Marine-Derived Streptomyces sp.

    PubMed Central

    Bae, Munhyung; Park, So Hyun; Kwon, Yun; Lee, Sang Kook; Shin, Jongheon; Nam, Joo-Won; Oh, Dong-Chan

    2017-01-01

    Based on profiles of secondary metabolites produced by marine bacteria obtained using LC/MS, succinilenes A–D (1–4), new triene polyols, were discovered from a culture of a Streptomyces strain SAK1, which was collected in the southern area of Jeju Island, Republic of Korea. The gross structures of 1–4 were primarily determined through analysis of NMR spectra. The double bond geometries of the succinilenes, which could not be established from conventional 1H NMR spectra because of the highly overlapped olefinic signals, were successfully deciphered using the recently developed quantum-mechanics-driven 1H iterative full spin analysis (QM-HiFSA). Succinilenes A–C (1–3) displayed inhibitory effects against lipopolysaccharide (LPS)-induced nitric oxide (NO) production, indicating their anti-inflammatory significance. These three compounds (1–3) commonly bear a succinic acid moiety, although succinilene D (4), which did not inhibit NO production, does not have this moiety in its structure. The absolute configurations of succinilenes A–D (1–4) were established through J-based configuration analysis, the modified Mosher’s method following methanolysis, and CD spectral analysis. PMID:28216577

  4. Differential proteomic analysis of an engineered Streptomyces coelicolor strain reveals metabolic pathways supporting growth on n-hexadecane.

    PubMed

    Gallo, Giuseppe; Lo Piccolo, Luca; Renzone, Giovanni; La Rosa, Ruggero; Scaloni, Andrea; Quatrini, Paola; Puglia, Anna Maria

    2012-06-01

    The alkB gene, encoding an alkane monooxygenase in the actinomycete Gordonia sp. SoCg, was expressed in the non-alkane-degrading actinomycete Streptomyces coelicolor M145. The resulting engineered strain, M145-AH, can grow on n-hexadecane as sole carbon source. To unravel proteins associated with growth on n-alkanes, proteome of M145-AH after 6, 24, and 48 h of incubation in the Bushnell-Haas (BH) mineral medium containing n-hexadecane as sole carbon source (H condition) and in BH without any carbon source (0 condition) were compared using 2D-differential gel electrophoresis. Proteome analysis revealed significant changes only at 48 h, showing 48 differentially abundant proteins identified by mass spectrometry procedures. To asses if these proteins were specifically related to n-hexadecane metabolism, their expression was investigated, comparing H proteome with that of M145-AH incubated in BH with glucose as sole carbon source (G condition). Thus, protein expression profiles at 6, 24, and 48 h under H, 0, and G conditions were combined, revealing that M145-AH regulates in a temporally- and carbon source-dependent manner the expression of proteins involved in regulatory events, central carbon metabolism, respiration, β-oxidation, membrane transport, and amino acid and protein metabolism. Interestingly, 21 % of them, mostly involved in membrane transport and protein metabolism, showed a n-hexadecane-dependent regulation with regulatory proteins such as CRP likely to have a key role in M145-AH n-hexadecane growth. These results, expanding the knowledge on n-alkane utilization in Gram-positive bacteria, reveal genes to be targeted to develop an efficient S. coelicolor M145-AH-based bioremediation system.

  5. Binding and molecular dynamic studies of sesquiterpenes (2R-acetoxymethyl-1,3,3-trimethyl-4t-(3-methyl-2-buten-1-yl)-1t-cyclohexanol) derived from marine Streptomyces sp. VITJS8 as potential anticancer agent.

    PubMed

    Naine, S Jemimah; Devi, C Subathra; Mohanasrinivasan, V; Doss, C George Priya; Kumar, D Thirumal

    2016-03-01

    The main aim of the current study is to explore the bioactive potential of Streptomyces sp. VITJS8 isolated from the marine saltern. The cultural, biochemical, and morphological studies were performed to acquire the characteristic features of the potent isolate VITJS8. The 16Sr DNA sequencing was performed to investigate the phylogenetic relationship between the Streptomyces genera. The structure of the compound was elucidated by gas chromatography-mass spectrometry (GC-MS), infra-red (IR), and ultra-violet (UV) spectroscopic data analysis. The GC-MS showed the retention time at 22.39 with a single peak indicating the purity of the active compound, and the molecular formula was established as C14H9ONCl2 based on the peak at m/z 277 [M](+). Furthermore, separated by high-performance liquid chromatography (HPLC), their retention time (t r) 2.761 was observed with the absorption maxima at 310 nm. The active compound showed effective inhibitory potential against four clinical pathogens at 500 μg/mL. The antioxidant activity was found effective at the IC50 value of 500 μg/mL with 90 % inhibition. The 3-(4,5-dimethylthiazol-2-yl)-2,5-ditetrazolium bromide (MTT) assay revealed the cytotoxicity against HepG2 cells at IC50 of 250 μg/mL. The progression of apoptosis was evidenced by morphological changes by nuclear staining. The DNA fragmentation pattern was observed at 250 μg/mL concentration. Based on flow cytometric analysis, it was evident that the compound was effective in inhibiting the sub-G0/G1 phase of cell cycle. The in vitro findings were also supported by the binding mode molecular docking studies. The active compound revealed minimum binding energy of -7.84 and showed good affinity towards the active region of topoisomerase-2α that could be considered as a suitable inhibitor. Lastly, we performed 30 ns molecular dynamic simulation analysis using GROMACS to aid in better designing of anticancer drugs. Simulation result of root mean square deviation (RMSD

  6. Biotransformation of trinitrotoluene (TNT) by Streptomyces species

    SciTech Connect

    Funk, S.B.; Pasti-Grigsby, M.B.; Felicione, E.C.; Crawford, D.L.

    1995-12-31

    Composting has been proposed as one process for use in the bioremediation of 2,4,6 trinitrotoluene (TNT)-contaminated soils. However, the biotransformations of TNT that occur during composting, and the specific compost microorganisms involved in TNT metabolism, are not well understood. Both mesophilic and thermophilic actinomycetes are important participants in the biodegradation of organic matter, and possibly TNT, in composts. Here the authors report on the biotransformation of TNT by Streptomyces species growing aerobically in a liquid medium supplemented with 10 to 100 mg/L of TNT. Streptomyces spp. are able to completely remove TNT from the culture medium within 24 hours. As has been observed with other bacteria, these streptomycetes transform TNT first by reducing the 4-nitro and 2-nitro groups to the corresponding amino group; reducing TNT first to 4-amino-2,6-dinitrotoluene and then 2,4-diamino-6-nitrotoluene. These intermediates are transitory and are themselves removed from the medium within 7 days.

  7. Cloning and Heterologous Expression of a Large-sized Natural Product Biosynthetic Gene Cluster in Streptomyces Species

    PubMed Central

    Nah, Hee-Ju; Pyeon, Hye-Rim; Kang, Seung-Hoon; Choi, Si-Sun; Kim, Eung-Soo

    2017-01-01

    Actinomycetes family including Streptomyces species have been a major source for the discovery of novel natural products (NPs) in the last several decades thanks to their structural novelty, diversity and complexity. Moreover, recent genome mining approach has provided an attractive tool to screen potentially valuable NP biosynthetic gene clusters (BGCs) present in the actinomycetes genomes. Since many of these NP BGCs are silent or cryptic in the original actinomycetes, various techniques have been employed to activate these NP BGCs. Heterologous expression of BGCs has become a useful strategy to produce, reactivate, improve, and modify the pathways of NPs present at minute quantities in the original actinomycetes isolates. However, cloning and efficient overexpression of an entire NP BGC, often as large as over 100 kb, remain challenging due to the ineffectiveness of current genetic systems in manipulating large NP BGCs. This mini review describes examples of actinomycetes NP production through BGC heterologous expression systems as well as recent strategies specialized for the large-sized NP BGCs in Streptomyces heterologous hosts. PMID:28360891

  8. Siderophore production by actinomycetes isolates from two soil sites in Western Australia.

    PubMed

    Lee, Joanna; Postmaster, Armin; Soon, Hooi Peng; Keast, David; Carson, Kerry C

    2012-04-01

    The actinomycetes are metabolically flexible soil micro-organisms capable of producing a range of compounds of interest, including siderophores. Siderophore production by actinomycetes sampled from two distinct and separate geographical sites in Western Australia were investigated and found to be generally similar in the total percentage of siderophore producers found. The only notable difference was the proportion of isolates producing catechol siderophores with only 3% found in site 1 (from the north-west of Western Australia and reportedly containing 40% magnetite) and 17% in site 2 (a commercial stone fruit orchard in the hills east of Perth with a soil base ranging from sandy loam to laterite). Further detailed characterization of isolates of interest identified a Streptomyces that produced extracellularly excreted enterobactin, the characteristic Enterobacteriaceae siderophore, and also revealed some of the conditions required for enterobactin production. Carriage of the entF gene, which codes for the synthetase responsible for the final assembly of the tri-cyclic structure of enterobactin, was confirmed by PCR in this isolate. Another separate Streptomyces produced a compound that matched the UV/VIS spectra of heterobactin, a siderophore previously only described in Rhodococcus and Nocardia.

  9. Multiplexed site-specific genome engineering for overproducing bioactive secondary metabolites in actinomycetes.

    PubMed

    Li, Lei; Zheng, Guosong; Chen, Jun; Ge, Mei; Jiang, Weihong; Lu, Yinhua

    2017-03-01

    Actinomycetes produce a large variety of pharmaceutically active compounds, yet production titers often require to be improved for discovery, development and large-scale manufacturing. Here, we describe a new technique, multiplexed site-specific genome engineering (MSGE) via the 'one integrase-multiple attB sites' concept, for the stable integration of secondary metabolite biosynthetic gene clusters (BGCs). Using MSGE, we achieved five-copy chromosomal integration of the pristinamycin II (PII) BGC in Streptomyces pristinaespiralis, resulting in the highest reported PII titers in flask and batch fermentations (2.2 and 2g/L, respectively). Furthermore, MSGE was successfully extended to develop a panel of powerful Streptomyces coelicolor heterologous hosts, in which up to four copies of the BGCs for chloramphenicol or anti-tumour compound YM-216391 were efficiently integrated in a single step, leading to significantly elevated productivity (2-23 times). Our multiplexed approach holds great potential for robust genome engineering of industrial actinomycetes and novel drug discovery by genome mining.

  10. Genome sequencing reveals complex secondary metabolome in themarine actinomycete Salinispora tropica

    SciTech Connect

    Udwary, Daniel W.; Zeigler, Lisa; Asolkar, Ratnakar; Singan,Vasanth; Lapidus, Alla; Fenical, William; Jensen, Paul R.; Moore, BradleyS.

    2007-05-01

    Recent fermentation studies have identified actinomycetes ofthe marine-dwelling genus Salinispora as prolific natural productproducers. To further evaluate their biosynthetic potential, we analyzedall identifiable secondary natural product gene clusters from therecently sequenced 5,184,724 bp S. tropica CNB-440 circular genome. Ouranalysis shows that biosynthetic potential meets or exceeds that shown byprevious Streptomyces genome sequences as well as other naturalproduct-producing actinomycetes. The S. tropica genome features ninepolyketide synthase systems of every known formally classified family,non-ribosomal peptide synthetases and several hybrid clusters. While afew clusters appear to encode molecules previously identified inStreptomyces species,the majority of the 15 biosynthetic loci are novel.Specific chemical information about putative and observed natural productmolecules is presented and discussed. In addition, our bioinformaticanalysis was critical for the structure elucidation of the novelpolyenemacrolactam salinilactam A. This study demonstrates the potentialfor genomic analysis to complement and strengthen traditional naturalproduct isolation studies and firmly establishes the genus Salinispora asa rich source of novel drug-like molecules.

  11. Isolation and characterization of Cr(VI)-reducing actinomycetes from estuarine sediments.

    PubMed

    Terahara, Takeshi; Xu, Xudan; Kobayashi, Takeshi; Imada, Chiaki

    2015-04-01

    Bioremediation technologies have strong potential use in the less costly and more environmentally friendly removal of highly toxic hexavalent-chromium (Cr(VI)) compared with physicochemical technologies. Several Cr(VI)-reducing bacteria have been isolated; however, there are few studies on Cr(VI)-resistant and Cr(VI)-reducing actinomycetes. In this study, Cr(VI)-reducing actinomycetes were screened from estuarine, marine, and terrestrial samples on the basis of Cr(VI)-resistant and Cr(VI)-reducing ability. Of the 80 Streptomyces-like strains isolated, 20 strains were found to be resistant to 50 mg/l of Cr(VI). In addition, two strains isolated from the estuarine sediment of Tokyo Bay were found to be resistant to a concentration of 150 mg/l of Cr(VI). Furthermore, one Cr(VI)-reducing strain was found to remove 60 mg/l of Cr(VI) within 1 week and was identified as Streptomyces thermocarboxydus based on 16S rRNA gene analysis. The comparative evaluation with the type strain S. thermocarboxydus NBRC 16323 showed that our isolated strain had higher ability to grow at 27 °C and reduce Cr(VI) at a NaCl concentration of 6.0 % at 27 °C compared with the type strain NBRC 16323. These results indicate that our isolated strain have a potential ability to remove Cr(VI) from contaminated, highly saline sources without heating.

  12. Screening of Marine Actinomycetes from Segara Anakan for Natural Pigment and Hydrolytic Activities

    NASA Astrophysics Data System (ADS)

    Asnani, A.; Ryandini, D.; Suwandri

    2016-02-01

    Marine actinomycetes have become sources of great interest to natural product chemistry due to their new chemical entities and bioactive metabolites. Since April 2010, we have screened actinobacteria from five sites that represent different ecosystems of Segara Anakan lagoon. In this present study we focus on specific isolates, K-2C which covers 1) actinomycetes identification based on morphology observation and 16S rRNA gene; 2) fermentation and isolation of pigment; 3) structure determination of pigment; and 4) hydrolytic enzymes characterization; Methodologies relevant to the studies were implemented accordingly. The results indicated that K-2C was likely Streptomyces fradiae strain RSU15, and the best fermentation medium should contain starch and casein with 21 days of incubation. The isolate has extracellular as well as intracellular pigments. Isolated pigments gave purple color with λmax of 529.00 nm. The pigment was structurally characterized. Interestingly, Streptomyces K-2C was able to produce potential hydrolytic enzymes such as amylase, cellulase, protease, lipase, urease, and nitrate reductase.

  13. Biosurfactant produced from Actinomycetes nocardiopsis A17: Characterization and its biological evaluation.

    PubMed

    Chakraborty, Samrat; Ghosh, Mandakini; Chakraborti, Srijita; Jana, Sougata; Sen, Kalyan Kumar; Kokare, Chandrakant; Zhang, Lixin

    2015-08-01

    This investigation aims to isolate an Actinomycetes strain producing a biosurfactant from the unexplored region of industrial and coal mine areas. Actinomycetes are selected for this study as their novel chemistry was not exhausted and they have tremendous potential to produce bioactive secondary metabolites. The biosurfactant was characterized and further needed to be utilized for pharmaceutical dosage form. Isolation, purification, screening, and characterization of the Actinomycetes A17 were done followed by its fermentation in optimized conditions. The cell-free supernatant was used for the extraction of the biosurfactant and precipitated by cold acetone. The dried precipitate was purified by TLC and the emulsification index, surface tension and CMC were determined. The isolated strain with preferred results was identified as Actinomycetes nocardiopsis A17 with high foam-forming properties. It gives lipase, amylase, gelatinase, and protease activity. The emulsification index was found to be 93±0.8 with surface tension 66.67 dyne/cm at the lowest concentration and cmc 0.6 μg/ml. These biosurfactants were characterized by Fourier transform infra red (FT-IR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS). Therefore, it can be concluded that the biosurfactant produced by Actinomycetes nocardiopsis sp. strain A17 was found to have satisfactory results with high surface activity and emulsion-forming ability.

  14. [Distribution and characteristics of soil antagonistic actinomycetes on northern slope of Taibai Mountain, Qinling].

    PubMed

    Zhu, Wen-Jie; Xue, Quan-Hong; Cao, Yan-Ru; Xue, Lei; Shen, Guang-Hui; Lai, Hang-Xian

    2011-11-01

    Twelve representative soil samples were collected from different altitudes on the northern slope of Taibai Mountain to study the distribution and characteristics of soil antagonistic actinomyces by using agar block method. There existed a great deal of soil antagonistic actinomyces in the study area. Among the 141 actinomycete strains isolated, 116 strains (82.3%) showed antagonism toward 12 target bacteria or fungi. The antagonistic strains at altitudes 800-1845, 3488, 3655, and 3670 m occupied 73.7% -86.8%, 81.3%, 78.9% and 82.3% of the total, respectively. 42.1% of the strains at altitudes 1200-2300 m and > 3400 m showed strong and broad spectrum antagonistic activity, suggesting that there was a great potential for the isolation of actinomycete strains with strong anti-biotic capability at these altitudes. 24.1% of the antagonistic actinomycetes showed antagonism against Staphyloccocus aureu, and 2.4%, 6.9% and 11.2% of them showed activity toward Verticillium dahliae in cotton, Phytophthora sp. in strawberry and Neonectria radiciccla in ginseng, respectively. This study showed that the soil actinomycete antagonistic potentiality (SAAP) could be used as a quantitative indicator to evaluate the potential of antagonistic actinomycete resources in soil.

  15. Chromium(VI) resistance and removal by actinomycete strains isolated from sediments.

    PubMed

    Polti, Marta A; Amoroso, María J; Abate, Carlos M

    2007-03-01

    Forty-one isolated actinomycetes were used to study qualitative and semi-quantitative screening of chromium(VI) resistance. Chromate-removing activity was estimated using the Cr(VI) specific colorimetric reagent 1,5-diphenylcarbazide. Twenty percent of the isolates from El Cadillal (EC) and 14% of isolates from a copper filter plant (CFP) were able to grow at 13 mM of Cr(VI). All isolates from sugar cane (SCP) could grow up to Cr(VI) concentration of 17 mM. EC, CFP and SCP strains were able to remove 24%, 30% and more than 40% of Cr(VI), respectively. The highest and lowest Cr(VI) specific removal values were 75.5 mg g(-1) cell by M3 (CFP), and 1.5 mg g(-1) cell by C35 (EC) strains. Eleven Cr(VI) resistant strains were characterized and identified as species of the genera Streptomyces (10) and Amycolatopsis (1). Differences on actinomycete community composition between contaminated and non-contaminated soil were found. This study showed the potential capacity of actinomycetes as tools for Cr(VI) bioremediation.

  16. [Coculture of actinomycetes with Bacillus subtilis and its effect on the bioactive secondary metabolites].

    PubMed

    Huang, Bing; Liu, Ning; Huang, Ying; Chen, Jinchun

    2009-06-01

    To explore the effect of coculturing actinomycetes with Bacillus subtilis on the production of bioactive secondary metabolites, we studied the difference between fermentation products of monocultures and the corresponding cocultures of 22 actinomycetes by antimicrobial assay and HPLC-PDA analysis. We selected Streptomyces strain FXJ2.014 with high bioactivity for further analysis and found additional metabolites in fermentation extracts of cocultures of strains FXJ2.014, FXJ1.296 and AS 4.1252 respectively with B. subtilis. Quinomycin A was the main bioactive metabolite produced by the monoculture of strain FXJ2.014, while a new quinomycin-like component named FXJ2.014-HB was produced when strain FXJ2.014 was cocultured with B. subtilis. Further tests of antimicrobial and antitumor activities indicated that FXJ2.014-HB and Quinomycin A had significant differences in terms of bioactivity. Moreover, the inhibitory activity of FXJ2.014-HB to a variety of tumor cell lines was weaker than the highly toxic Quinomycin A, indicating its potential to be an antibiotic with low cell toxicity. In conclusion, coculture can be used as a promising approach to discover bioactive secondary metabolites from actinomycetes.

  17. I-SceI endonuclease: a new tool for DNA repair studies and genetic manipulations in streptomyces.

    PubMed

    Siegl, Theresa; Petzke, Lutz; Welle, Elisabeth; Luzhetskyy, Andriy

    2010-07-01

    Actinomycetes are Gram-positive bacteria with a complex life cycle. They produce many pharmaceutically relevant secondary metabolites, including antibiotics and anticancer drugs. However, there is a limited number of biotechnological applications available as opposed to genetic model organisms like Bacillus subtilis or Escherichia coli. We report here a system for the functional expression of a synthetic gene encoding the I-SceI homing endonuclease in several streptomycetes. Using the synthetic sce(a) gene, we were able to create controlled genomic DNA double-strand breaks. A mutagenesis system, based on the homing endonuclease I-SceI, has been developed to construct targeted, non-polar, unmarked gene mutations in Streptomyces sp. Tü6071. In addition, we have shown that homologous recombination is a major pathway in streptomycetes to repair an I-SceI-generated DNA double-strand break. This novel I-SceI-based tool will be useful in fundamental studies on the repair mechanism of DNA double-strand breaks and for a variety of biotechnological applications.

  18. Latex Clearing Protein (Lcp) of Streptomyces sp. Strain K30 Is a b-Type Cytochrome and Differs from Rubber Oxygenase A (RoxA) in Its Biophysical Properties.

    PubMed

    Birke, Jakob; Röther, Wolf; Jendrossek, Dieter

    2015-06-01

    Specific polyisoprene-cleaving activities of 1.5 U/mg and 4.6 U/mg were determined for purified Strep-tagged latex clearing protein (Lcp) of Streptomyces sp. strain K30 at 23 °C and 37 °C, respectively. Metal analysis revealed the presence of approximately one atom of iron per Lcp molecule. Copper, which had been identified in Lcp1VH2 of Gordonia polyisoprenivorans previously, was below the detection limit in LcpK30. Heme was identified as a cofactor in purified LcpK30 by (i) detection of characteristic α-, β-, and γ (Soret)-bands at 562 nm, 532 nm, and 430 nm in the visible spectrum after chemical reduction, (ii) detection of an acetone-extractable porphyrin molecule, (iii) determination of a heme b-type-specific absorption maximum (556 nm) after chemical conversion of the heme group to a bipyridyl-heme complex, and (iv) detection of a b-heme-specific m/z value of 616.2 via mass spectrometry. Spectroscopic analysis showed that purified Lcp as isolated contains an oxidized heme-Fe(3+) that is free of bound dioxygen. This is in contrast to the rubber oxygenase RoxA, a c-type heme-containing polyisoprene-cleaving enzyme present in Gram-negative rubber degraders, in which the covalently bound heme firmly binds a dioxygen molecule. LcpK30 also differed from RoxA in the lengths of the rubber degradation cleavage products and in having a higher melting point of 61.5 °C (RoxA, 54.3 °C). In summary, RoxA and Lcp both are equipped with a heme cofactor and catalyze an oxidative C-C cleavage reaction but differ in the heme subgroup type and in several biochemical and biophysical properties. These findings suggest differences in the catalytic reaction mechanisms.

  19. Cloning and Characterization of 1-Deoxy-d-Xylulose 5-Phosphate Synthase from Streptomyces sp. Strain CL190, Which Uses both the Mevalonate and Nonmevalonate Pathways for Isopentenyl Diphosphate Biosynthesis

    PubMed Central

    Kuzuyama, Tomohisa; Takagi, Motoki; Takahashi, Shunji; Seto, Haruo

    2000-01-01

    In addition to the ubiquitous mevalonate pathway, Streptomyces sp. strain CL190 utilizes the nonmevalonate pathway for isopentenyl diphosphate biosynthesis. The initial step of this nonmevalonate pathway is the formation of 1-deoxy-d-xylulose 5-phosphate (DXP) by condensation of pyruvate and glyceraldehyde 3-phosphate catalyzed by DXP synthase. The corresponding gene, dxs, was cloned from CL190 by using PCR with two oligonucleotide primers synthesized on the basis of two highly conserved regions among dxs homologs from six genera. The dxs gene of CL190 encodes 631 amino acid residues with a predicted molecular mass of 68 kDa. The recombinant enzyme overexpressed in Escherichia coli was purified as a soluble protein and characterized. The molecular mass of the enzyme was estimated to be 70 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 130 kDa by gel filtration chromatography, suggesting that the enzyme is most likely to be a dimer. The enzyme showed a pH optimum of 9.0, with a Vmax of 370 U per mg of protein and Kms of 65 μM for pyruvate and 120 μM for d-glyceraldehyde 3-phosphate. The purified enzyme catalyzed the formation of 1-deoxyxylulose by condensation of pyruvate and glyceraldehyde as well, with a Km value of 35 mM for d-glyceraldehyde. To compare the enzymatic properties of CL190 and E. coli DXP synthases, the latter enzyme was also overexpressed and purified. Although these two enzymes had different origins, they showed the same enzymatic properties. PMID:10648511

  20. Diversity of Streptomyces spp. in Eastern Himalayan region – computational RNomics approach to phylogeny

    PubMed Central

    Bhattacharjee, Kaushik; Banerjee, Subhro; Joshi, Santa Ram

    2012-01-01

    Isolation and characterization of actinomycetes from soil samples from altitudinal gradient of North-East India were investigated for computational RNomics based phylogeny. A total of 52 diverse isolates of Streptomyces from the soil samples were isolated on four different media and from these 6 isolates were selected on the basis of cultural characteristics, microscopic and biochemical studies. Sequencing of 16S rDNA of the selected isolates identified them to belong to six different species of Streptomyces. The molecular morphometric and physico-kinetic analysis of 16S rRNA sequences were performed to predict the diversity of the genus. The computational RNomics study revealed the significance of the structural RNA based phylogenetic analysis in a relatively diverse group of Streptomyces. PMID:22829729

  1. Renaissance in antibacterial discovery from actinomycetes.

    PubMed

    Baltz, Richard H

    2008-10-01

    The soil actinomycetes have been important sources of antibiotics, but were nearly abandoned in recent years in favor of high-throughput target-based screening of chemical libraries. The latter approach has not been productive, so it is time to reinvigorate the discovery of new antibiotics from a proven source. Recent progress has been made on antibiotic discovery from actinomycetes by using high-throughput fermentation, isolation of marine actinomycetes, mining genomes for cryptic pathways, and combinatorial biosynthesis to generate new secondary metabolites related to existing pharmacophores.

  2. Activation and products of the cryptic secondary metabolite biosynthetic gene clusters by rifampin resistance (rpoB) mutations in actinomycetes.

    PubMed

    Tanaka, Yukinori; Kasahara, Ken; Hirose, Yutaka; Murakami, Kiriko; Kugimiya, Rie; Ochi, Kozo

    2013-07-01

    A subset of rifampin resistance (rpoB) mutations result in the overproduction of antibiotics in various actinomycetes, including Streptomyces, Saccharopolyspora, and Amycolatopsis, with H437Y and H437R rpoB mutations effective most frequently. Moreover, the rpoB mutations markedly activate (up to 70-fold at the transcriptional level) the cryptic/silent secondary metabolite biosynthetic gene clusters of these actinomycetes, which are not activated under general stressful conditions, with the exception of treatment with rare earth elements. Analysis of the metabolite profile demonstrated that the rpoB mutants produced many metabolites, which were not detected in the wild-type strains. This approach utilizing rifampin resistance mutations is characterized by its feasibility and potential scalability to high-throughput studies and would be useful to activate and to enhance the yields of metabolites for discovery and biochemical characterization.

  3. Study of the diversity of culturable actinomycetes in the North Pacific and Caribbean coasts of Costa Rica

    PubMed Central

    Solano, Godofredo; Rojas-Jiménez, Keilor; Jaspars, Marcel

    2011-01-01

    In this study, 137 actinomycetes were isolated from subtidal marine sediments in the North Pacific and Caribbean coasts of Costa Rica. Bioinformatics analysis of the 16S rRNA gene sequences assigned the isolates to 15 families and 21 genera. Streptomyces was the dominant genus while the remaining 20 genera were poorly represented. Nearly 70% of the phylotypes presented a coastal-restricted distribution whereas the other 30% were common inhabitants of both shores. The coastal tropical waters of Costa Rica showed a high diversity of actinomycetes, both in terms of the number of species and phylogenetic composition, although significant differences were observed between and within shores. The observed pattern of species distribution might be the result of several factors including the characteristics of the ecosystems, presence of endemic species and the influence of terrestrial runoff. PMID:19365710

  4. Antitumor Compounds from Marine Actinomycetes

    PubMed Central

    Olano, Carlos; Méndez, Carmen; Salas, José A.

    2009-01-01

    Chemotherapy is one of the main treatments used to combat cancer. A great number of antitumor compounds are natural products or their derivatives, mainly produced by microorganisms. In particular, actinomycetes are the producers of a large number of natural products with different biological activities, including antitumor properties. These antitumor compounds belong to several structural classes such as anthracyclines, enediynes, indolocarbazoles, isoprenoides, macrolides, non-ribosomal peptides and others, and they exert antitumor activity by inducing apoptosis through DNA cleavage mediated by topoisomerase I or II inhibition, mitochondria permeabilization, inhibition of key enzymes involved in signal transduction like proteases, or cellular metabolism and in some cases by inhibiting tumor-induced angiogenesis. Marine organisms have attracted special attention in the last years for their ability to produce interesting pharmacological lead compounds. PMID:19597582

  5. Isolation of cellulolytic actinomycetes from marine sediments

    SciTech Connect

    Veiga, M.; Esparis, A.; Fabregas, J.

    1983-07-01

    The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity. 13 references.

  6. Acidophilic actinomycetes from rhizosphere soil: diversity and properties beneficial to plants.

    PubMed

    Poomthongdee, Nalin; Duangmal, Kannika; Pathom-aree, Wasu

    2015-02-01

    Three hundred and fifty-one isolates of actinomycetes were recovered from 21 rhizospheric soil samples using acidified media of pH 5.5. They were evaluated for their antifungal, siderophore production and phosphate solubilization activities. The total count of actinomycetes growing on acidified starch casein agar and Gause no. 1 agar were below 2.48 × 10(4) CFU g(-1) soil. Two hundred and twelve isolates were assigned to acidophiles and the remaining 139 isolates were neutrophiles. Of these actinomycetes, 57.8, 32.5 and 50.4%, showed antagonistic activity against three rice pathogenic fungi; Fusarium moniliforme, Helminthosporium oryzae and Rhizoctonia solani, respectively. More than half of the isolates (68.1%) inhibited at least one tested pathogenic fungus, whereas 25.9% exhibited antifungal activities against all tested fungi. Three hundred and thirty-eight isolates (96.3%) produced siderophore and 266 isolates (75.8%) solubilized phosphate. A greater proportion of the acidophilic actinomycetes exhibited antifungal, siderophore production and phosphate solubilization activity compared with the neutrophiles. Three hundred and twenty-five isolates (92.6%) were classified as streptomycetes based on their morphological characteristics and the presence of the LL-isomeric form of diaminopimelic acid in whole-cell hydrolysates. The 16S ribosomal RNA (rRNA) gene analysis of representative non-streptomycete strains showed that the isolates belonged to seven genera, that is, Allokutzneria, Amycolatopsis, Mycobacterium, Nocardia, Nonomuraea, Saccharopolyspora and Verrucosispora. The potential antifungal acidophilic isolates, R9-4, R14-1, R14-5 and R20-5, showed close similarity to Streptomyces misionensis NBRC 13063(T) (AB184285) in terms of morphological characteristics and 16S rRNA gene sequences.

  7. Distribution and generic composition of culturable marine actinomycetes from the sediments of Indian continental slope of Bay of Bengal

    NASA Astrophysics Data System (ADS)

    Das, Surajit; Lyla, P. S.; Ajmal Khan, S.

    2008-05-01

    Actinomycetes population from continental slope sediment of the Bay of Bengal was studied. Samples were collected during two voyages of FORV Sagar Sampada in 2004 (May-June) and 2005 (July) respectively from 11 transects (each transect had ca. 200 m, 500 m, and 1 000 m depth stations). The physicochemical parameters of overlying water, and sediment samples were also recorded. The actinomycete population ranged from 5.17 to 51.94 CFU/g dry sediment weight and 9.38 to 45.22 CFU/g dry sediment weight during the two cruises respectively. No actinomycete colony was isolated from stations in 1 000 m depth. Two-way analysis of variance showed significant variation among stations (ANOVA two-way, P<0.05), but no significance was found between the two cruises (ANOVA two-way, P<0.05). Populations in stations in 500 m depth in both cruises were higher than that of 200 m depth stations with statistically insignificant difference (ANOVA two-way, P>0.05). Three actinomycetes genera were identified. Streptomyces was found to be the dominating one in both the cruises, followed by Micromonospora, and Actinomyces. The spore of Streptomyces isolates showed the abundance in spiral spore chain. Spore surface was smooth. Multiple regression analysis revealed that the influencing physico-chemical factors were sediment pH, sediment temperature, TOC, porosity, salinity, and pressure. The media used in the present study was prepared with seawater. Thus, they may represent an autochthonous marine flora and deny the theory of land runoff carriage into the sea for adaptation to the salinity of the seawater and sediments.

  8. Elicitation of secondary metabolism in actinomycetes.

    PubMed

    Abdelmohsen, Usama Ramadan; Grkovic, Tanja; Balasubramanian, Srikkanth; Kamel, Mohamed Salah; Quinn, Ronald J; Hentschel, Ute

    2015-11-01

    Genomic sequence data have revealed the presence of a large fraction of putatively silent biosynthetic gene clusters in the genomes of actinomycetes that encode for secondary metabolites, which are not detected under standard fermentation conditions. This review focuses on the effects of biological (co-cultivation), chemical, as well as molecular elicitation on secondary metabolism in actinomycetes. Our review covers the literature until June 2014 and exemplifies the diversity of natural products that have been recovered by such approaches from the phylum Actinobacteria.

  9. An Efficient Method To Generate Gene Deletion Mutants of the Rapamycin-Producing Bacterium Streptomyces iranensis HM 35

    PubMed Central

    Netzker, Tina; Schroeckh, Volker; Gregory, Matthew A.; Flak, Michal; Krespach, Mario K. C.; Leadlay, Peter F.

    2016-01-01

    ABSTRACT Streptomyces iranensis HM 35 is an alternative rapamycin producer to Streptomyces rapamycinicus. Targeted genetic modification of rapamycin-producing actinomycetes is a powerful tool for the directed production of rapamycin derivatives, and it has also revealed some key features of the molecular biology of rapamycin formation in S. rapamycinicus. The approach depends upon efficient conjugational plasmid transfer from Escherichia coli to Streptomyces, and the failure of this step has frustrated its application to Streptomyces iranensis HM 35. Here, by systematically optimizing the process of conjugational plasmid transfer, including screening of various media, and by defining optimal temperatures and concentrations of antibiotics and Ca2+ ions in the conjugation media, we have achieved exconjugant formation for each of a series of gene deletions in S. iranensis HM 35. Among them were rapK, which generates the starter unit for rapamycin biosynthesis, and hutF, encoding a histidine catabolizing enzyme. The protocol that we have developed may allow efficient generation of targeted gene knockout mutants of Streptomyces species that are genetically difficult to manipulate. IMPORTANCE The developed protocol of conjugational plasmid transfer from Escherichia coli to Streptomyces iranensis may allow efficient generation of targeted gene knockout mutants of other genetically difficult to manipulate, but valuable, Streptomyces species. PMID:27037115

  10. Antibacterial agents from actinomycetes - a review.

    PubMed

    Mahajan, Girish Badrinath; Balachandran, Lakshmi

    2012-01-01

    The discovery of Penicillin in 1928 and that of Streptomycin in 1943, has been pivotal to the exploration of nature as a source of new lead molecules. Globally, the microbiologist today is acknowledged as a crucial player in the drug discovery program. The microbial products, especially those from actinomycetes have been a phenomenal success for the past seven decades. Bioprospecting for new leads are often compounded by the recurrence of known antibiotics in newer microbial isolates. Despite all these deterrents, actinomycetes have proved to be a sustained mine of novel antibiotics, which selectively destroys the pathogens without affecting the host tissues. Each of these antibiotics is unique in their mode of action. Their versatility and immense economic value is something, which is extremely noteworthy. The anti-infective turn-over of over 79 billion US dollars in 2009, includes about 166 antibiotics and derivatives such as the Beta-lactam peptide antibiotics, the macrolide polyketide erythromycin, tetracyclines, aminoglycosides, daptomycin, tigecycline, most of which are produced by actinomycetes (1). Actinomycetes continue to play a highly significant role in drug discovery and development. Among the bioactive compounds that have been obtained so far from microbes, 45 % are produced by actinomycetes, 38 % by fungi and 17 % by unicellular eubacteria (2). Further many chemically synthesized drugs owe their origin to natural sources. In this review article, we highlight the recent antibiotics from actinomycetes with emphasis on their source, structures, activity, mechanism of action and current status.

  11. Case report of Streptomyces endocarditis of a prosthetic aortic valve.

    PubMed Central

    Mossad, S B; Tomford, J W; Stewart, R; Ratliff, N B; Hall, G S

    1995-01-01

    We describe the first case of prosthetic valve endocarditis due to a Streptomyces sp. The patient presented with fever, cutaneous embolic lesions, and bacteremia 3 months after aortic valve replacement. Treatment required valve replacement and a long course of parenteral imipenem. PMID:8586732

  12. Bioactive potential of Streptomyces against fish and shellfish pathogens

    PubMed Central

    Selvakumar, D; Arun, K; Suguna, S; Kumar, D; Dhevendaran, K

    2010-01-01

    Background and Objectives In the present study, isolation of Streptomyces associated with marine sponges and its bioactive potential against fish and shellfish pathogens were assessed. The Streptomyces sp. were isolated from the marine sponges namely Callyspongia diffusa, Mycale mytilorum, Tedania anhelans and Dysidea fragilis collected from Vizhinjam port, situated in the South-West coast of India. Materials and Methods The Streptomyces associated with marine sponges were isolated using specific ISP media. The isolates of Streptomyces were characterized for their colony characteristics, morphological properties, physiological and biochemical properties and were tentatively identified. The strains were cultivated on a lab scale level as shake-flask cultures and the crude extracts of the bioactive compounds obtained with ethyl acetate were screened biologically and chemically. By biological screening, the extracts were analyzed for their activity against fish and shellfish pathogens namely Aeromonas hydrophila, Serratia sp. and Vibrio spp, using the disk and agar-well diffusion bioassay method, while by chemical screening the crude culture extracts were analyzed by TLC and UV–Vis spectrophotometer. Results Ninety-four isolates were found to be associated with marine sponges, among them only seven strains showed antagonism against fish and shellfish pathogens. Analysis of morphological, physiological and biochemical characteristics suggested that these strains belonged to the genus Streptomyces. The initial screening of the isolates by spot inoculation method exhibited antibacterial activity against Aeromonas hydrophila. In-vitro screening of the submerge culture extracts showed positive inhibition against the fish and shellfish pathogens namely Aeromonas hydrophila, Serratia sp. and Vibrio spp. The screening of bioactive compounds confirmed the production of polyene substances by UV spectrum, which resulted in absorbance peaks ranging from 225 to 245 nm and TLC

  13. The Synthesis of Quinolone Natural Products from Pseudonocardia sp.

    PubMed Central

    Salvaggio, Flavia; Hodgkinson, James T.; Carro, Laura; Geddis, Stephen M.; Galloway, Warren R. J. D.; Welch, Martin

    2015-01-01

    Abstract The synthesis of four quinolone natural products from the actinomycete Pseudonocardia sp. is reported. The key step involved a sp2–sp3 Suzuki–Miyaura reaction between a common boronic ester lateral chain and various functionalised quinolone cores. The quinolones slowed growth of E. coli and S. aureus by inducing extended lag phases.

  14. Marine actinomycete diversity and natural product discovery.

    PubMed

    Jensen, Paul R; Mincer, Tracy J; Williams, Philip G; Fenical, William

    2005-01-01

    Microbial natural products remain an important resource for drug discovery yet the microorganisms inhabiting the world's oceans have largely been overlooked in this regard. The recent discovery of novel secondary metabolites from taxonomically unique populations of marine actinomycetes suggests that these bacteria add an important new dimension to microbial natural product research. Continued efforts to characterize marine actinomycete diversity and how adaptations to the marine environment affect secondary metabolite production will create a better understanding of the potential utility of these bacteria as a source of useful products for biotechnology.

  15. Effect of crude extracts of selected actinomycetes on biofilm formation of A. schindleri, M. aci, and B. cereus.

    PubMed

    Saleem, Hafiz Ghulam Murtaza; Aftab, Usman; Sajid, Imran; Abbas, Zaigham; Sabri, Anjum Nasim

    2015-05-01

    Actinomycetes are well known group of gram positive bacteria for their potential to produce antibiotics. This study sought to assess the ability of the selected actinomycetes to control biofilm forming bacteria isolated from different dental plaque samples. On the basis of morphological differences three out of ten different dental plaque bacterial isolates were selected for further study. These isolates were biochemically and genetically characterized and were identified as Acinetobacter schinndleri, Moraxella aci, and Bacillus cereus. Antibiotic resistant profile was measured through disc diffusion method and found that all three isolates were moderately sensitive to ofloxacin and erythromycin and resistant to trimethoprim. Antibacterial activity of ten different Streptomyces strains was assessed through an agar plug and well diffusion method against three dental biofilm forming bacteria. Two Streptomyces strains named as S. erythrogriseus and S. labedae showed good antibacterial activity against Moraxella and Acinetobacter strains. Ability of the four active antibiotic producing strains to inhibit biofilm formation was assessed using microtiter biofilm detection assay. It was found that biofilm forming ability of Acinetobacter and Moraxella was inhibited by S. labedae an antibiotic producing strain, while S. macrosporeus can only inhibit biofilm formation by B. cereus.

  16. Effect of antibiotic down-regulatory gene wblA ortholog on antifungal polyene production in rare actinomycetes Pseudonocardia autotrophica.

    PubMed

    Kim, Hye-Jin; Kim, Min-Kyung; Jin, Ying-Yu; Kim, Young-Woo; Kim, Eung-Soo

    2014-09-01

    The rare actinomycete Pseudonocardia autotrophica was previously shown to produce a solubilityimproved toxicity-reduced novel polyene compound named Nystatin-like Pseudonocardia Polyene (NPP). The low productivity of NPP in P. autotrophica implies that its biosynthetic pathway is tightly regulated. In this study, wblApau was isolated and identified as a novel negative regulatory gene for NPP production in P. autotrophica, which showed approximately 49% amino acid identity with a global antibiotic down-regulatory gene, wblA, identified from various Streptomycetes species. Although no significant difference in NPP production was observed between P. autotrophica harboring empty vector and the S. coelicolor wblA under its native promoter, approximately 12% less NPP was produced in P. autotrophica expressing the wblA gene under the strong constitutive ermE(*) promoter. Furthermore, disruption of the wblApau gene from P. autotrophica resulted in an approximately 80% increase in NPP productivity. These results strongly suggest that identification and inactivation of the global antibiotic down-regulatory gene wblA ortholog are a critical strategy for improving secondary metabolite overproduction in not only Streptomyces but also non-Streptomyces rare actinomycete species.

  17. Production of specialized metabolites by Streptomyces coelicolor A3(2).

    PubMed

    van Keulen, Geertje; Dyson, Paul J

    2014-01-01

    The actinomycetes are well-known bioactive natural product producers, comprising the Streptomycetes, the richest drug-prolific family in all kingdoms, producing therapeutic compounds for the areas of infection, cancer, circulation, and immunity. Completion and annotation of many actinomycete genomes has highlighted further how proficient these bacteria are in specialized metabolism, which have been largely underexploited in traditional screening programs. The genome sequence of the model strain Streptomyces coelicolor A3(2), and subsequent development of genomics-driven approaches to understand its large specialized metabolome, has been key in unlocking the high potential of specialized metabolites for natural product genomics-based drug discovery. This review discusses systematically the biochemistry and genetics of each of the specialized metabolites of S. coelicolor and describes metabolite transport processes for excretion and complex regulatory patterns controlling biosynthesis.

  18. Novel tryptophan metabolism by a potential gene cluster that is widely distributed among actinomycetes.

    PubMed

    Ozaki, Taro; Nishiyama, Makoto; Kuzuyama, Tomohisa

    2013-04-05

    The characterization of potential gene clusters is a promising strategy for the identification of novel natural products and the expansion of structural diversity. However, there are often difficulties in identifying potential metabolites because their biosynthetic genes are either silenced or expressed only at a low level. Here, we report the identification of a novel metabolite that is synthesized by a potential gene cluster containing an indole prenyltransferase gene (SCO7467) and a flavin-dependent monooxygenase (FMO) gene (SCO7468), which were mined from the genome of Streptomyces coelicolor A3(2). We introduced these two genes into the closely related Streptomyces lividans TK23 and analyzed the culture broths of the transformants. This process allowed us to identify a novel metabolite, 5-dimethylallylindole-3-acetonitrile (5-DMAIAN) that was overproduced in the transformant. Biochemical characterization of the recombinant SCO7467 and SCO7468 demonstrated the novel L-tryptophan metabolism leading to 5-DMAIAN. SCO7467 catalyzes the prenylation of L-tryptophan to form 5-dimethylallyl-L-tryptophan (5-DMAT). This enzyme is the first actinomycetes prenyltransferase known to catalyze the addition of a dimethylallyl group to the C-5 of tryptophan. SCO7468 then catalyzes the conversion of 5-DMAT into 5-dimethylallylindole-3-acetaldoxime (5-DMAIAOx). An aldoxime-forming reaction catalyzed by the FMO enzyme was also identified for the first time in this study. Finally, dehydration of 5-DMAIAOx presumably occurs to yield 5-DMAIAN. This study provides insight into the biosynthesis of prenylated indoles that have been purified from actinomycetes.

  19. Evolutionary relationships among actinophages and a putative adaptation for growth in Streptomyces spp.

    PubMed

    Smith, Margaret C M; Hendrix, Roger W; Dedrick, Rebekah; Mitchell, Kaitlin; Ko, Ching-Chung; Russell, Daniel; Bell, Emma; Gregory, Matthew; Bibb, Maureen J; Pethick, Florence; Jacobs-Sera, Deborah; Herron, Paul; Buttner, Mark J; Hatfull, Graham F

    2013-11-01

    The genome sequences of eight Streptomyces phages are presented, four of which were isolated for this study. Phages R4, TG1, Hau3, and SV1 were isolated previously and have been exploited as tools for understanding and genetically manipulating Streptomyces spp. We also extracted five apparently intact prophages from recent Streptomyces spp. genome projects and, together with six phage genomes in the database, we analyzed all 19 Streptomyces phage genomes with a view to understanding their relationships to each other and to other actinophages, particularly the mycobacteriophages. Fifteen of the Streptomyces phages group into four clusters of related genomes. Although the R4-like phages do not share nucleotide sequence similarity with other phages, they clearly have common ancestry with cluster A mycobacteriophages, sharing many protein homologues, common gene syntenies, and similar repressor-stoperator regulatory systems. The R4-like phage Hau3 and the prophage StrepC.1 (from Streptomyces sp. strain C) appear to have hijacked a unique adaptation of the streptomycetes, i.e., use of the rare UUA codon, to control translation of the essential phage protein, the terminase. The Streptomyces venezuelae generalized transducing phage SV1 was used to predict the presence of other generalized transducing phages for different Streptomyces species.

  20. Culturable rare Actinomycetes: diversity, isolation and marine natural product discovery.

    PubMed

    Subramani, Ramesh; Aalbersberg, William

    2013-11-01

    Rare Actinomycetes from underexplored marine environments are targeted in drug discovery studies due to the Actinomycetes' potentially huge resource of structurally diverse natural products with unusual biological activity. Of all marine bacteria, 10 % are Actinomycetes, which have proven an outstanding and fascinating resource for new and potent bioactive molecules. Past and present efforts in the isolation of rare Actinomycetes from underexplored diverse natural habitats have resulted in the isolation of about 220 rare Actinomycete genera of which more than 50 taxa have been reported to be the producers of 2,500 bioactive compounds. That amount represents greater than 25 % of the total Actinomycetes metabolites, demonstrating that selective isolation methods are being developed and extensively applied. Due to the high rediscovery rate of known compounds from Actinomycetes, a renewed interest in the development of new antimicrobial agents from rare and novel Actinomycetes is urgently required to combat the increasing number of multidrug-resistant human pathogens. To facilitate that discovery, this review updates all selective isolation media including pretreatment and enrichment methods for the isolation of marine rare Actinomycetes. In addition, this review demonstrates that discovering new compounds with novel scaffolds can be increased by intensive efforts in isolating and screening rare marine genera of Actinomycetes. Between 2007 and mid-2013, 80 new rare Actinomycete species were reported from marine habitats. They belong to 23 rare families, of which three are novel, and 20 novel genera. Of them, the family Micromonosporaceae is dominant as a producer of promising chemical diversity.

  1. Systematic and biotechnological aspects of halophilic and halotolerant actinomycetes.

    PubMed

    Hamedi, Javad; Mohammadipanah, Fatemeh; Ventosa, Antonio

    2013-01-01

    More than 70 species of halotolerant and halophilic actinomycetes belonging to at least 24 genera have been validly described. Halophilic actinomycetes are a less explored source of actinomycetes for discovery of novel bioactive secondary metabolites. Degradation of aliphatic and aromatic organic compounds, detoxification of pollutants, production of new enzymes and other metabolites such as antibiotics, compatible solutes and polymers are other potential industrial applications of halophilic and halotolerant actinomycetes. Especially new bioactive secondary metabolites that are derived from only a small fraction of the investigated halophilic actinomycetes, mainly from marine habitats, have revealed the huge capacity of this physiological group in production of new bioactive chemical entities. Combined high metabolic capacities of actinomycetes and unique features related to extremophilic nature of the halophilic actinomycetes have conferred on them an influential role for future biotechnological applications.

  2. Structural and Functional Characterizations of SsgB, a Conserved Activator of Developmental Cell Division in Morphologically Complex Actinomycetes*

    PubMed Central

    Xu, Qingping; Traag, Bjørn A.; Willemse, Joost; McMullan, Daniel; Miller, Mitchell D.; Elsliger, Marc-André; Abdubek, Polat; Astakhova, Tamara; Axelrod, Herbert L.; Bakolitsa, Constantina; Carlton, Dennis; Chen, Connie; Chiu, Hsiu-Ju; Chruszcz, Maksymilian; Clayton, Thomas; Das, Debanu; Deller, Marc C.; Duan, Lian; Ellrott, Kyle; Ernst, Dustin; Farr, Carol L.; Feuerhelm, Julie; Grant, Joanna C.; Grzechnik, Anna; Grzechnik, Slawomir K.; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K.; Klock, Heath E.; Knuth, Mark W.; Kozbial, Piotr; Krishna, S. Sri; Kumar, Abhinav; Marciano, David; Minor, Wladek; Mommaas, A. Mieke; Morse, Andrew T.; Nigoghossian, Edward; Nopakun, Amanda; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Puckett, Christina; Reyes, Ron; Rife, Christopher L.; Sefcovic, Natasha; Tien, Henry J.; Trame, Christine B.; van den Bedem, Henry; Wang, Shuren; Weekes, Dana; Hodgson, Keith O.; Wooley, John; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.; van Wezel, Gilles P.

    2009-01-01

    SsgA-like proteins (SALPs) are a family of homologous cell division-related proteins that occur exclusively in morphologically complex actinomycetes. We show that SsgB, a subfamily of SALPs, is the archetypal SALP that is functionally conserved in all sporulating actinomycetes. Sporulation-specific cell division of Streptomyces coelicolor ssgB mutants is restored by introduction of distant ssgB orthologues from other actinomycetes. Interestingly, the number of septa (and spores) of the complemented null mutants is dictated by the specific ssgB orthologue that is expressed. The crystal structure of the SsgB from Thermobifida fusca was determined at 2.6 Å resolution and represents the first structure for this family. The structure revealed similarities to a class of eukaryotic “whirly” single-stranded DNA/RNA-binding proteins. However, the electro-negative surface of the SALPs suggests that neither SsgB nor any of the other SALPs are likely to interact with nucleotide substrates. Instead, we show that a conserved hydrophobic surface is likely to be important for SALP function and suggest that proteins are the likely binding partners. PMID:19567872

  3. Spontaneous and induced mutations to rifampicin, streptomycin and spectinomycin resistances in actinomycetes: mutagenic mechanisms and applications for strain improvement.

    PubMed

    Baltz, Richard H

    2014-09-01

    Chemical mutagenesis continues to be an important foundational methodology for the generation of highly productive actinomycete strains for the commercial production of antibiotics and other secondary metabolites. In the past, the determination of frequencies of chemically induced resistance to rifampicin (RifR), spectinomycin (SpcR) and streptomycin (StrR) have served as surrogate markers to monitor the efficiencies and robustness of mutagenic protocols. Recent studies indicate that high level RifR, SpcR and StrR phenotypes map to specific regions of the rpoB, rpsE and rpsL genes, respectively, in actinomycetes. Moreover, mutagenesis to RifR can occur spontaneously at many different sites in rpoB, and all six types of base-pair substitutions, as well as in-frame deletions and insertions, have been observed. The RifR/rpoB system provides a robust method to rank mutagenic protocols, to evaluate mutagen specificity and to study spontaneous mutagenesis mechanisms involved in the maintenance of high G+C content in Streptomyces species and other actinomycetes.

  4. Structural and Functional Characterizations of SsgB, a Conserved Activator of Developmental Cell Division in Morphologically Complex Actinomycetes

    SciTech Connect

    Xu, Qingping; Traag, Bjørn A.; Willemse, Joost; McMullan, Daniel; Miller, Mitchell D.; Elsliger, Marc-André; Abdubek, Polat; Astakhova, Tamara; Axelrod, Herbert L.; Bakolitsa, Constantina; Carlton, Dennis; Chen, Connie; Chiu, Hsiu-Ju; Chruszcz, Maksymilian; Clayton, Thomas; Das, Debanu; Deller, Marc C.; Duan, Lian; Ellrott, Kyle; Ernst, Dustin; Farr, Carol L.; Feuerhelm, Julie; Grant, Joanna C.; Grzechnik, Anna; Grzechnik, Slawomir K.; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K.; Klock, Heath E.; Knuth, Mark W.; Kozbial, Piotr; Krishna, S. Sri; Kumar, Abhinav; Marciano, David; Minor, Wladek; Mommaas, A. Mieke; Morse, Andrew T.; Nigoghossian, Edward; Nopakun, Amanda; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Puckett, Christina; Reyes, Ron; Rife, Christopher L.; Sefcovic, Natasha; Tien, Henry J.; Trame, Christine B.; van den Bedem, Henry; Wang, Shuren; Weekes, Dana; Hodgson, Keith O.; Wooley, John; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.; van Wezel, Gilles P.

    2010-01-20

    SsgA-like proteins (SALPs) are a family of homologous cell division-related proteins that occur exclusively in morphologically complex actinomycetes. We show that SsgB, a subfamily of SALPs, is the archetypal SALP that is functionally conserved in all sporulating actinomycetes. Sporulation-specific cell division of Streptomyces coelicolor ssgB mutants is restored by introduction of distant ssgB orthologues from other actinomycetes. Interestingly, the number of septa (and spores) of the complemented null mutants is dictated by the specific ssgB orthologue that is expressed. The crystal structure of the SsgB from Thermobifida fusca was determined at 2.6 {angstrom} resolution and represents the first structure for this family. The structure revealed similarities to a class of eukaryotic 'whirly' single-stranded DNA/RNA-binding proteins. However, the electro-negative surface of the SALPs suggests that neither SsgB nor any of the other SALPs are likely to interact with nucleotide substrates. Instead, we show that a conserved hydrophobic surface is likely to be important for SALP function and suggest that proteins are the likely binding partners.

  5. Study of the cellulases produced by three mesophilic actinomycetes grown on bagasse as substrate

    SciTech Connect

    Van Zyl, W.H.

    1985-09-01

    The cellulases that strains of Streptomyces albogrisolus, S. nitrosporeus, and Micromonospora melanosporea produce when grown on untreated ballmilled bagasse were investigated. Optimum conditions for extracellular cellulase production and activity were determined to be growth at pH 6.7-7.4 and 25-35 degrees C for 4-5 days and assay at pH 5.0-6.0 and 45-55 degrees C, respectively. The endoglucanases were thermally stable at 50 degrees C, but the Avicelases had a half-life of approximately 24 hours at this temperature. Nearly half of the endoglucanases and almost all of the Avicelases were absorbed on ballmilled bagasse after 15 minutes incubation at 50 degrees C. The ..beta..-glucosidases were found to be mainly intracellular or cell wall bound. These mesophilic actinomycetes concomitantly produced xylanases and ..beta..-xylosidases with cellulases that, apart from cellobiose and glucose, also release xylose from bagasse. This feature may be advantageous in the commercial application of the enzymes of mesophilic actinomycetes for the saccharification of natural cellulosic substrates.

  6. Structural and functional properties of actinomycetal communities in chernozems and saline soils of Ukraine

    NASA Astrophysics Data System (ADS)

    Grishko, V. N.; Syshchikova, O. V.

    2010-02-01

    In the profiles of ordinary and southern chernozems, the total numbers of amylolytic microorganisms and actinomycetes decreased with the depth by 2.4-4.2 and 3.4 times, respectively; in the profiles of solonetz and solonchak soils, by 4.2-5.3 and 4.8 times, respectively. In the genetic horizons of the ordinary and southern chernozems, the share of actinomycetes amounted to 29-30% of the total population of microorganisms; in the saline soils, it increases with the depth from 23 to 43%. In the chernozems, Streptomyces violaceomaculatus (Roseus section), St. sporoherbeus (Azureus), St. aerionidulus (Cinereus), St. enduracidicus (Cinereus), and St. grisinus (Cinereus) predominated; in the saline soils, St. violaceomaculatus and St. aerionidulus prevailed. In the ordinary chernozem, the Berger-Parker index was 1.5 times higher than in the southern chernozem. High similarity was found between the streptomycete communities in the chernozems (the Sorensen coefficient was 0.78). In the solonetzes, the species richness of the streptomycetes was higher by 1.7 times than in the solonchaks. In the chernozems, the similarity of the streptomycete communities was higher than in the solonchaks (0.78 and 0.60, respectively).

  7. A diaminopimelic acid auxotrophic Escherichia coli donor provides improved counterselection following intergeneric conjugation with actinomycetes.

    PubMed

    Allard, Nancy; Garneau, Daniel; Poulin-Laprade, Dominic; Burrus, Vincent; Brzezinski, Ryszard; Roy, Sébastien

    2015-08-01

    Considering the medical, biotechnological, and economical importance of actinobacteria, there is a continuous need to improve the tools for genetic engineering of a broad range of these microorganisms. Intergeneric conjugation has proven to be a valuable yet imperfect tool for this purpose. The natural resistance of many actinomycetes to nalidixic acid (Nal) is generally exploited to eliminate the sensitive Escherichia coli donor strain following conjugation. Nevertheless, Nal can delay growth and have other unexpected effects on the recipient strain. To provide an improved alternative to antibiotics, we propose a postconjugational counterselection using a diaminopimelic acid (DAP) auxotrophic donor strain. The DAP-negative phenotype was obtained by introducing a dapA deletion into the popular methylase-negative donor strain E. coli ET12567/pUZ8002. The viability of ET12567 and its ΔdapA mutant exposed to DAP deprivation or Nal selection were compared in liquid pure culture and after mating with Streptomyces coelicolor. Results showed that death of the E. coli ΔdapA Nal-sensitive donor strain occurred more efficiently when subjected to DAP deprivation than when exposed to Nal. Our study shows that postconjugational counterselection based on DAP deprivation circumvents the use of antibiotics and will facilitate the transfer of plasmids into actinomycetes with high biotechnological potential, yet currently not accessible to conjugative techniques.

  8. Development of actinomycetes in brown semidesert soil under low water pressure

    NASA Astrophysics Data System (ADS)

    Zvyagintsev, D. G.; Zenova, G. M.; Sudnitsyn, I. I.; Gracheva, T. A.; Lapygina, E. E.; Napol'skaya, K. R.; Sydnitsyna, A. E.

    2012-07-01

    Under laboratory conditions, the spores of a xerotolerant Streptomyces odorifera strain germinated in brown semidesert soil even at extremely low soil water pressure ( P = -96.4 MPa, -964 atm, a w 0.50); the plantlets increased in length and formed mycelium, on which a new generation of spores was produced (a complete development cycle of the actinomycetes—from a spore to the formation of new spores—passed). The duration of the first cycles of the actinomycetes' development varied from 13 days at P = -27 atm to 57 days at P = -964 atm and was directly proportional to the absolute value of the soil water pressure ( P). In the first cycles of the actinomycetes' development, the rate of increase of the concentration of the germinated spores and mycelium, as well as the logarithms of the mycelium-to-germinated spore concentration ratios, was inversely proportional to the logarithm of P. These relationships indicated that the energy state of the water determined its availability to soil biota and, hence, the activity of its physiological and biochemical processes.

  9. Antimicrobial potential of Halophilic actinomycetes against multi drug resistant (MDR) ventilator associated pneumonia causing bacterial pathogens.

    PubMed

    Aslam, Sana; Sajid, Imran

    2016-03-01

    A collection of forty halophilic actinomycetes isolated from water and mud samples of the saline lake at Kalar Kahar, salt range, Pakistan, was screened to investigate their antimicrobial potential against multi drug resistant (MDR) ventilator associated pneumonia causing bacterial pathogens. The isolates exhibited significant tolerance to alkaline conditions and grew well at pH 9-11. The taxonomic status of the isolated strains was determined by morphological, biochemical and physiological characterization and by 16s rRNA gene sequencing. The results revealed that majority of the isolates (90%) belong to the genus Streptomyces. Most of the isolates exhibited remarkable antimicrobial activity up to 20mm zone of inhibition against MDR ventilator associated pneumonia causing bacteria including Staphylococcus aureus, Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Enterobacter and Acinetobacter spp. Additionally the isolates showed moderate to high cytotoxicity in the range of 40 to 80% larval mortality against Artemia salina in a micro well cytotoxicity assay. The chemical screening or the so called metabolic fingerprinting of the methanolic extracts of each isolate, by thin layer chromatography (TLC) using various staining reagents and by high performance liquid chromatography (HPLC-UV), indicated an impressive diversity of the compounds produced by these strains. The study reveals that these halophilic actinomycetes are a promising source of bioactive compounds. The preparative scale fermentation, isolation, purification and structure elucidation of the compounds produced by them may yield novel antimicrobial or chemotherapeutic agents.

  10. Comparative genomics of actinomycetes with a focus on natural product biosynthetic genes

    PubMed Central

    2013-01-01

    Background Actinomycetes are a diverse group of medically, industrially and ecologically important bacteria, studied as much for the diseases they cause as for the cures they hold. The genomes of actinomycetes revealed that these bacteria have a large number of natural product gene clusters, although many of these are difficult to tie to products in the laboratory. Large scale comparisons of these clusters are difficult to perform due to the presence of highly similar repeated domains in the most common biosynthetic machinery: polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs). Results We have used comparative genomics to provide an overview of the genomic features of a set of 102 closed genomes from this important group of bacteria with a focus on natural product biosynthetic genes. We have focused on well-represented genera and determine the occurrence of gene cluster families therein. Conservation of natural product gene clusters within Mycobacterium, Streptomyces and Frankia suggest crucial roles for natural products in the biology of each genus. The abundance of natural product classes is also found to vary greatly between genera, revealing underlying patterns that are not yet understood. Conclusions A large-scale analysis of natural product gene clusters presents a useful foundation for hypothesis formulation that is currently underutilized in the field. Such studies will be increasingly necessary to study the diversity and ecology of natural products as the number of genome sequences available continues to grow. PMID:24020438

  11. In vitro actinomycete biofilm development and inhibition by the polyene antibiotic, nystatin, on IUD copper surfaces.

    PubMed

    Shanmughapriya, Santhanam; Francis, Arumugam Lency; Kavitha, Senthil; Natarajaseenivasan, Kalimuthusamy

    2012-01-01

    The presence of intrauterine contraceptive devices (IUDs) gives a solid surface for attachment and an ideal niche for biofilm to form and flourish. Pelvic actinomycosis is often associated with the use of IUDs. Treatment of IUD-associated pelvic actinomycosis requires the immediate removal of the IUD. Therefore, this article presents in vitro evidence to support the use of novel antibiotics in the treatment of actinomycete biofilms. Twenty one clinical actinomycetes isolates from endocervical swabs of IUD wearers were assessed for their biofilm forming ability. An in vitro biofilm model with three isolates, Streptomyces strain A4, Nocardia strain C15 and Nocardia strain C17 was subjected to treatment with nystatin. Inhibition of biofilm formation by nystatin was found to be concentration dependent, with MBIC50 values in the range 0.08-0.16 mg ml(-1). Furthermore, at a concentration of 0.16 mg ml(-1), nystatin inhibited the twitching motility of the isolates, providing evidence for a possible mechanism of biofilm inhibition.

  12. Streptomyces rhizobacteria modulate the secondary metabolism of Eucalyptus plants.

    PubMed

    Salla, Tamiris Daros; da Silva, Ramos; Astarita, Leandro Vieira; Santarém, Eliane Romanato

    2014-12-01

    The genus Eucalyptus comprises economically important species, such as Eucalyptus grandis and Eucalyptus globulus, used especially as a raw material in many industrial sectors. Species of Eucalyptus are very susceptible to pathogens, mainly fungi, which leads to mortality of plant cuttings in rooting phase. One alternative to promote plant health and development is the potential use of microorganisms that act as agents for biological control, such as plant growth-promoting rhizobacteria (PGPR). Rhizobacteria Streptomyces spp have been considered as PGPR. This study aimed at selecting strains of Streptomyces with ability to promote plant growth and modulate secondary metabolism of E. grandis and E. globulus in vitro plants. The experiments assessed the development of plants (root number and length), changes in key enzymes in plant defense (polyphenol oxidase and peroxidase) and induction of secondary compounds(total phenolic and quercetinic flavonoid fraction). The isolate Streptomyces PM9 showed highest production of indol-3-acetic acid and the best potential for root induction. Treatment of Eucalyptus roots with Streptomyces PM9 caused alterations in enzymes activities during the period of co-cultivation (1-15 days), as well as in the levels of phenolic compounds and flavonoids. Shoots also showed alteration in the secondary metabolism, suggesting induced systemic response. The ability of Streptomyces sp. PM9 on promoting root growth, through production of IAA, and possible role on modulation of secondary metabolism of Eucalyptus plants characterizes this isolate as PGPR and indicates its potential use as a biological control in forestry.

  13. Actinomycetes: A Source of Lignocellulolytic Enzymes

    PubMed Central

    Saini, Anita; Aggarwal, Neeraj K.; Sharma, Anuja; Yadav, Anita

    2015-01-01

    Lignocellulose is the most abundant biomass on earth. Agricultural, forest, and agroindustrial activities generate tons of lignocellulosic wastes annually, which present readily procurable, economically affordable, and renewable feedstock for various lignocelluloses based applications. Lignocelluloses are the focus of present decade researchers globally, in an attempt to develop technologies based on natural biomass for reducing dependence on expensive and exhaustible substrates. Lignocellulolytic enzymes, that is, cellulases, hemicellulases, and lignolytic enzymes, play very important role in the processing of lignocelluloses which is prerequisite for their utilization in various processes. These enzymes are obtained from microorganisms distributed in both prokaryotic and eukaryotic domains including bacteria, fungi, and actinomycetes. Actinomycetes are an attractive microbial group for production of lignocellulose degrading enzymes. Various studies have evaluated the lignocellulose degrading ability of actinomycetes, which can be potentially implemented in the production of different value added products. This paper is an overview of the diversity of cellulolytic, hemicellulolytic, and lignolytic actinomycetes along with brief discussion of their hydrolytic enzyme systems involved in biomass modification. PMID:26793393

  14. Degradation by Streptomyces viridosporus T7A of plant material grown under elevated CO2 conditions.

    PubMed

    Ball, A S

    1991-11-15

    The biodegradability of plant material derived from wheat grown under different concentrations of atmospheric CO2 was investigated using the lignocarbohydrate solubilising actinomycete, Streptomyces viridosporus. Growth of S. viridosporus and solubilisation of lignocarbohydrate were highest when wheat grown at ambient CO2 concentrations (350 ppm) was used as C-source. Growth of S. viridosporus and solubilisation were reduced when the plant material was derived from wheat grown at 645 ppm CO2. The results suggest that modifications in plant structure occur when wheat is grown under conditions of elevated atmospheric CO2 which make it more resistant to microbial digestion.

  15. Copper removal ability by Streptomyces strains with dissimilar growth patterns and endowed with cupric reductase activity.

    PubMed

    Albarracín, Virginia Helena; Avila, Ana Lucía; Amoroso, María Julia; Abate, Carlos Mauricio

    2008-11-01

    Morphological, physiological and molecular characterization of three copper-resistant actinobacterial strains (AB2A, AB3 and AB5A) isolated from copper-polluted sediments of a drainage channel showed that they belonged to the genus Streptomyces. These characteristics plus their distinctive copper resistance phenotypes revealed considerable divergence among the isolates. Highly dissimilar growth patterns and copper removal efficiency were observed for the selected Streptomyces strains grown on minimal medium (MM) added with 0.5 mM of copper sulfate (MM(Cu)). Strain AB2A showed an early mechanism of copper uptake/retention (80% until day 3), followed by a drastic metal efflux process (days 5-7). In contrast, Streptomyces sp. AB3 and AB5A showed only copper retention phenotypes under the same culture conditions. Particularly, Streptomyces sp. AB5A showed a better efficiency in copper removal (94%), although a longer lag phase was observed for this microorganism grown for 7 days in MM(Cu). Cupric reductase activity was detected in both copper-adapted cells and nonadapted cells of all three strains but this activity was up to 100-fold higher in preadapted cells of Streptomyces sp. AB2A. To our knowledge, this is the first time that cupric reductase activity was demonstrated in Streptomyces strains.

  16. Comparative genomic hybridizations reveal absence of large Streptomyces coelicolor genomic islands in Streptomyces lividans

    PubMed Central

    Jayapal, Karthik P; Lian, Wei; Glod, Frank; Sherman, David H; Hu, Wei-Shou

    2007-01-01

    Background The genomes of Streptomyces coelicolor and Streptomyces lividans bear a considerable degree of synteny. While S. coelicolor is the model streptomycete for studying antibiotic synthesis and differentiation, S. lividans is almost exclusively considered as the preferred host, among actinomycetes, for cloning and expression of exogenous DNA. We used whole genome microarrays as a comparative genomics tool for identifying the subtle differences between these two chromosomes. Results We identified five large S. coelicolor genomic islands (larger than 25 kb) and 18 smaller islets absent in S. lividans chromosome. Many of these regions show anomalous GC bias and codon usage patterns. Six of them are in close vicinity of tRNA genes while nine are flanked with near perfect repeat sequences indicating that these are probable recent evolutionary acquisitions into S. coelicolor. Embedded within these segments are at least four DNA methylases and two probable methyl-sensing restriction endonucleases. Comparison with S. coelicolor transcriptome and proteome data revealed that some of the missing genes are active during the course of growth and differentiation in S. coelicolor. In particular, a pair of methylmalonyl CoA mutase (mcm) genes involved in polyketide precursor biosynthesis, an acyl-CoA dehydrogenase implicated in timing of actinorhodin synthesis and bldB, a developmentally significant regulator whose mutation causes complete abrogation of antibiotic synthesis belong to this category. Conclusion Our findings provide tangible hints for elucidating the genetic basis of important phenotypic differences between these two streptomycetes. Importantly, absence of certain genes in S. lividans identified here could potentially explain the relative ease of DNA transformations and the conditional lack of actinorhodin synthesis in S. lividans. PMID:17623098

  17. The biodegradation of layered silicates under the influence of cyanobacterial-actinomycetes associations

    NASA Astrophysics Data System (ADS)

    Ivanova, Ekaterina

    2013-04-01

    The weathering of sheet silicates is well known to be related to local and global geochemical cycles. Content and composition of clay minerals in soil determine the sorption properties of the soil horizons, water-holding capacity of the soil, stickiness, plasticity, etc. Microorganisms have a diverse range of mechanisms of minerals' structure transformation (acid- and alkali formation, biosorption, complexing, etc). One of the methods is an ability of exopolysaccharide-formation, in particular the formation of mucus, common to many bacteria, including cyanobacteria. Mucous covers cyanobacteria are the specific econiches for other bacteria, including actinomycetes. The objective was to analyze the structural changes of clay minerals under the influence of the cyanobacterial-actinomycetes associative growth. The objects of the study were: 1) the experimental symbiotic association, consisting of free-living heterocyst-formative cyanobacterium Anabaena variabilis Kutz. ATCC 294132 and actinomycete Streptomyces cyaneofuscatus FR837630, 2) rock samples obtained from the Museum of the Soil Science Department of the Lomonosov Moscow State University: kaolinite, consisting of kaolin (96%) Al4 (OH) 8 [Si4O10]; mixed with hydromica, chlorite and quartz; vermiculite, consisting of vermiculite (Ca, Mg, ...)*(Mg, Fe)3(OH)2[(Si, Al)4O10]*4H2O and trioctahedral mica (biotite). The mineralogical compositions of the rocks were determined by the universal X-ray Diffractometer Carl Zeiss Yena. The operationg regime was kept constant (30 kv, 40 mA). The cultivation of the association of actinomycete S. cyanoefuscatus and cyanobacterium A. variabilis caused a reduction in the intensity of kaolinite and hydromica reflexes. However, since both (mica and kaolinite) components have a rigid structure, the significant structural transformation of the minerals was not revealed. Another pattern was observed in the experiment, where the rock sample of vermiculite was used as the mineral

  18. Chromogenicity of Streptomyces.

    PubMed

    Arai, T; Mikami, Y

    1972-02-01

    A simplified technique to detect polyphenol oxidase and melanin formation by Streptomyces culture filtrates was developed. The procedure involves the direct assay of pigment formation by the culture filtrate with 3-(3,4-dihydroxyphenyl)-L-alanine (L-dopa) as a substrate. Among cultures of the International Streptomyces Project, 34 failed to produce a diffusible dark brown pigment on peptone-yeast extract-iron-agar and synthetic tyrosine-agar and gave a negative reaction to the melanin formation test. Sixteen cultures produced a diffusible dark brown pigment on both peptone-yeast extract-iron-agar and synthetic tyrosine-agar and gave positive reactions to the test with either L-tyrosine or L-dopa as substrate. Twenty-one cultures produced a diffusible dark brown pigment on peptone-yeast extract-iron-agar, but failed to do so on synthetic tyrosine-agar. Most of these cultures gave a positive reaction to the test when L-dopa was used as the substrate. The correlation between chromogenicity on complex organic media and melanin formation was more clearly established with L-dopa as substrate than with synthetic tyrosine-agar in the present test. The melanin formation test by the present technique, instead of chromogenicity on complex organic media, is recommended as a key feature for the classification of Streptomyces.

  19. Characterization of Streptomyces MITKK-103, a newly isolated actinomycin X2-producer.

    PubMed

    Kurosawa, K; Bui, V P; VanEssendelft, J L; Willis, L B; Lessard, P A; Ghiviriga, I; Sambandan, T G; Rha, C K; Sinskey, A J

    2006-08-01

    A new actinomycete strain designated MITKK-103 was isolated from the soil of a flowerpot using a humic acid agar medium. The newly isolated strain was able to produce a large amount of actinomycin X2 even under nonoptimized growing conditions and serves as a promising source of this antibiotic. Actinomycin X2 has higher cytotoxicity toward cultured human leukemia (HL-60) cells than does actinomycin D, and it induces cell death via apoptosis. A nearly complete 16S ribosomal DNA (rDNA) sequence from the isolate was determined and found to have high identity (98.5-100%) with Streptomyces galbus, Streptomyces griseofuscus, and Streptomyces padanus, indicating that MITKK-103 belongs to the genus Streptomyces. The isolate clustered with species belonging to the S. padanus clade in a 16S-rDNA-based phylogenetic tree and showed 75% overall homology to S. padanus ATCC 25646 in DNA-DNA relatedness analysis. Although the growth of the isolate was somewhat different from the three species mentioned, the strain MITKK-103 most closely resembles S. padanus on the basis of the morphological and phenotypic characteristics, phylogenetic analysis, and genotypic data. As such, this is the first report of a strain of S. padanus capable of producing actinomycins.

  20. Molecules to Ecosystems: Actinomycete Natural Products In situ

    PubMed Central

    Behie, Scott W.; Bonet, Bailey; Zacharia, Vineetha M.; McClung, Dylan J.; Traxler, Matthew F.

    2017-01-01

    Actinomycetes, filamentous actinobacteria found in numerous ecosystems around the globe, produce a wide range of clinically useful natural products (NP). In natural environments, actinomycetes live in dynamic communities where environmental cues and ecological interactions likely influence NP biosynthesis. Our current understating of these cues, and the ecological roles of NP, is in its infancy. We postulate that understanding the ecological context in which actinomycete metabolites are made is fundamental to advancing the discovery of novel NP. In this review we explore the ecological relevance of actinomycetes and their secondary metabolites from varying ecosystems, and suggest that investigating the ecology of actinomycete interactions warrants particular attention with respect to metabolite discovery. Furthermore, we focus on the chemical ecology and in situ analysis of actinomycete NP and consider the implications for NP biosynthesis at ecosystem scales. PMID:28144233

  1. Genome Content and Phylogenomics Reveal both Ancestral and Lateral Evolutionary Pathways in Plant-Pathogenic Streptomyces Species.

    PubMed

    Huguet-Tapia, Jose C; Lefebure, Tristan; Badger, Jonathan H; Guan, Dongli; Pettis, Gregg S; Stanhope, Michael J; Loria, Rosemary

    2016-01-29

    Streptomyces spp. are highly differentiated actinomycetes with large, linear chromosomes that encode an arsenal of biologically active molecules and catabolic enzymes. Members of this genus are well equipped for life in nutrient-limited environments and are common soil saprophytes. Out of the hundreds of species in the genus Streptomyces, a small group has evolved the ability to infect plants. The recent availability of Streptomyces genome sequences, including four genomes of pathogenic species, provided an opportunity to characterize the gene content specific to these pathogens and to study phylogenetic relationships among them. Genome sequencing, comparative genomics, and phylogenetic analysis enabled us to discriminate pathogenic from saprophytic Streptomyces strains; moreover, we calculated that the pathogen-specific genome contains 4,662 orthologs. Phylogenetic reconstruction suggested that Streptomyces scabies and S. ipomoeae share an ancestor but that their biosynthetic clusters encoding the required virulence factor thaxtomin have diverged. In contrast, S. turgidiscabies and S. acidiscabies, two relatively unrelated pathogens, possess highly similar thaxtomin biosynthesis clusters, which suggests that the acquisition of these genes was through lateral gene transfer.

  2. Genome Content and Phylogenomics Reveal both Ancestral and Lateral Evolutionary Pathways in Plant-Pathogenic Streptomyces Species

    PubMed Central

    Huguet-Tapia, Jose C.; Lefebure, Tristan; Badger, Jonathan H.; Guan, Dongli; Stanhope, Michael J.

    2016-01-01

    Streptomyces spp. are highly differentiated actinomycetes with large, linear chromosomes that encode an arsenal of biologically active molecules and catabolic enzymes. Members of this genus are well equipped for life in nutrient-limited environments and are common soil saprophytes. Out of the hundreds of species in the genus Streptomyces, a small group has evolved the ability to infect plants. The recent availability of Streptomyces genome sequences, including four genomes of pathogenic species, provided an opportunity to characterize the gene content specific to these pathogens and to study phylogenetic relationships among them. Genome sequencing, comparative genomics, and phylogenetic analysis enabled us to discriminate pathogenic from saprophytic Streptomyces strains; moreover, we calculated that the pathogen-specific genome contains 4,662 orthologs. Phylogenetic reconstruction suggested that Streptomyces scabies and S. ipomoeae share an ancestor but that their biosynthetic clusters encoding the required virulence factor thaxtomin have diverged. In contrast, S. turgidiscabies and S. acidiscabies, two relatively unrelated pathogens, possess highly similar thaxtomin biosynthesis clusters, which suggests that the acquisition of these genes was through lateral gene transfer. PMID:26826232

  3. Heterologous production of kasugamycin, an aminoglycoside antibiotic from Streptomyces kasugaensis, in Streptomyces lividans and Rhodococcus erythropolis L-88 by constitutive expression of the biosynthetic gene cluster.

    PubMed

    Kasuga, Kano; Sasaki, Akira; Matsuo, Takashi; Yamamoto, Chika; Minato, Yuiko; Kuwahara, Naoya; Fujii, Chikako; Kobayashi, Masayuki; Agematu, Hitosi; Tamura, Tomohiro; Komatsu, Mamoru; Ishikawa, Jun; Ikeda, Haruo; Kojima, Ikuo

    2017-02-27

    Kasugamycin (KSM), an aminoglycoside antibiotic isolated from Streptomyces kasugaensis cultures, has been used against rice blast disease for more than 50 years. We cloned the KSM biosynthetic gene (KBG) cluster from S. kasugaensis MB273-C4 and constructed three KBG cassettes (i.e., cassettes I-III) to enable heterologous production of KSM in many actinomycetes by constitutive expression of KBGs. Cassette I comprised all putative transcriptional units in the cluster, but it was placed under the control of the P neo promoter from Tn5. It was not maintained stably in Streptomyces lividans and did not transform Rhodococcus erythropolis. Cassette II retained the original arrangement of KBGs, except that the promoter of kasT, the specific activator gene for KBG, was replaced with P rpsJ , the constitutive promoter of rpsJ from Streptomyces avermitilis. To enhance the intracellular concentration of myo-inositol, an expression cassette of ino1 encoding the inositol-1-phosphate synthase from S. avermitilis was inserted into cassette II to generate cassette III. These two cassettes showed stable maintenance in S. lividans and R. erythropolis to produce KSM. Particularly, the transformants of S. lividans induced KSM production up to the same levels as those produced by S. kasugaensis. Furthermore, cassette III induced more KSM accumulation than cassette II in R. erythropolis, suggesting an exogenous supply of myo-inositol by the ino1 expression in the host. Cassettes II and III appear to be useful for heterologous KSM production in actinomycetes. Rhodococcus exhibiting a spherical form in liquid cultivation is also a promising heterologous host for antibiotic fermentation.

  4. Biocontrol of Rhizoctonia solani damping-off and promotion of tomato plant growth by endophytic actinomycetes isolated from native plants of Algerian Sahara.

    PubMed

    Goudjal, Yacine; Toumatia, Omrane; Yekkour, Amine; Sabaou, Nasserdine; Mathieu, Florence; Zitouni, Abdelghani

    2014-01-20

    Thirty-four endophytic actinomycetes were isolated from the roots of native plants of the Algerian Sahara. Morphological and chemical studies showed that twenty-nine isolates belonged to the Streptomyces genus and five were non-Streptomyces. All isolates were screened for their in vitro antifungal activity against Rhizoctonia solani. The six that had the greatest pathogen inhibitory capacities were subsequently tested for their in vivo biocontrol potential on R. solani damping-off in sterilized and non-sterilized soils, and for their plant-growth promoting activities on tomato seedlings. In both soils, coating tomato seeds with antagonistic isolates significantly reduced (P<0.05) the severity of damping-off of tomato seedlings. Among the isolates tested, the strains CA-2 and AA-2 exhibited the same disease incidence reduction as thioperoxydicarbonic diamide, tetramethylthiram (TMTD) and no significant differences (P<0.05) were observed. Furthermore, they resulted in a significant increase in the seedling fresh weight, the seedling length and the root length of the seed-treated seedlings compared to the control. The taxonomic position based on 16S rDNA sequence analysis and phylogenetic studies indicated that the strains CA-2 and AA-2 were related to Streptomyces mutabilis NBRC 12800(T) (100% of similarity) and Streptomyces cyaneofuscatus JCM 4364(T) (100% of similarity), respectively.

  5. Altered desferrioxamine-mediated iron utilization is a common trait of bald mutants of Streptomyces coelicolor.

    PubMed

    Lambert, Stéphany; Traxler, Matthew F; Craig, Matthias; Maciejewska, Marta; Ongena, Marc; van Wezel, Gilles P; Kolter, Roberto; Rigali, Sébastien

    2014-08-01

    Streptomyces coelicolor is an important model organism for developmental studies of filamentous GC-rich actinobacteria. The genetic characterization of mutants of S. coelicolor blocked at the vegetative mycelium stage, the so-called bald (bld) mutants that are unable to erect spore-forming aerial hyphae, has opened the way to discovering the molecular basis of development in actinomycetes. Desferrioxamine (DFO) production and import of ferrioxamines (FO; iron-complexed DFO) are key to triggering morphogenesis of S. coelicolor and we show here that growth of S. coelicolor on the reference medium for Streptomyces developmental studies is fully dependent on DFO biosynthesis. UPLC-ESI-MS analysis revealed that all bld mutants tested are affected in DFO biosynthesis, with bldA, bldJ, and ptsH mutants severely impaired in DFO production, while bldF, bldK, crr and ptsI mutants overproduce DFO. Morphogenesis of bldK and bldJ mutants was recovered by supplying exogenous iron. Transcript analysis showed that the bldJ mutant is impaired in expression of genes involved in the uptake of FO, whereas transcription of genes involved in both DFO biosynthesis and FO uptake is increased in bldK mutants. Our study allows proposing altered DFO production and/or FO uptake as a novel phenotypic marker of many S. coelicolor bld mutants, and strengthens the role of siderophores and iron acquisition in morphological development of actinomycetes.

  6. Marine actinomycetes: an ongoing source of novel bioactive metabolites.

    PubMed

    Subramani, Ramesh; Aalbersberg, William

    2012-12-20

    Actinomycetes are virtually unlimited sources of novel compounds with many therapeutic applications and hold a prominent position due to their diversity and proven ability to produce novel bioactive compounds. There are more than 22,000 known microbial secondary metabolites, 70% of which are produced by actinomycetes, 20% from fungi, 7% from Bacillus spp. and 1-2% by other bacteria. Among the actinomycetes, streptomycetes group are considered economically important because out of the approximately more than 10,000 known antibiotics, 50-55% are produced by this genus. The ecological role of actinomycetes in the marine ecosystem is largely neglected and various assumptions meant there was little incentive to isolate marine strains for search and discovery of new drugs. The search for and discovery of rare and new actinomycetes is of significant interest to drug discovery due to a growing need for the development of new and potent therapeutic agents. Modern molecular technologies are adding strength to the target-directed search for detection and isolation of bioactive actinomycetes, and continued development of improved cultivation methods and molecular technologies for accessing the marine environment promises to provide access to this significant new source of chemical diversity with novel/rare actinomycetes including new species of previously reported actinomycetes.

  7. Exploitation of phage battery in the search for bioactive actinomycetes.

    PubMed

    Kurtböke, D Ipek

    2011-02-01

    Screening of microbial natural products continues to represent an important route to the discovery of novel bioactive compounds for the development of new therapeutic agents, and actinomycetes are still the major producers of biopharmaceuticals. Selective isolation of bioactive actinomycete species, in particular the rare ones, has thus become a target for industrial microbiologists. In this context, bacteriophages have proven to be useful tools as (1) naturally present indicators of under-represented or rare actinomycete taxa in environmental samples, (2) indicators of the relatedness of bioactive taxa in target-directed search and discovery, (3) de-selection agents of unwanted taxa on isolation plates in target-specific search for rare actinomycete taxa, (4) tools in screening assays for specific targets. Against this background, a number of case studies are presented to illustrate the use of bacteriophages as tools in actinomycete-origin bioactive compound search and discovery programs.

  8. Screening and characterization of protease producing actinomycetes from marine saltern.

    PubMed

    Suthindhiran, Krish; Jayasri, Mangalam Achuthananda; Dipali, Dipa; Prasar, Apurva

    2014-10-01

    In the course of systematic screening program for bioactive actinomycetes, an alkaline protease producing halophilic strain Actinopolyspora sp. VITSDK2 was isolated from marine saltern, Southern India. The strain was identified as Actinopolyspora based on its phenotypic and phylogenetic characters. The protease was partially purified using ammonium sulfate precipitation and subsequently by DEAE cellulose column chromatography. The enzyme was further purified using HPLC and the molecular weight was found to be 22 kDa as determined by SDS-PAGE analysis. The purified protease exhibited pH stability in a wide range of 4-12 with optimum at 10.0. The enzyme was found to be stable between 25 and 80 °C and displayed a maximum activity at 60 °C. The enzyme activity was increased marginally in presence of Mn(2+) , Mg(2+) , and Ca(2+) and decreased in presence of Cu(2+) . PMSF and DFP completely inhibited the activity suggesting it belongs to serine protease. Further, the proteolytic activity was abolished in presence of N-tosyl-L-lysine chloromethyl ketone suggesting this might be chymotrypsin-like serine protease. The protease was 96% active when kept for 10 days at room temperature. The results indicate that the enzyme belong to chymotrypsin-like serine protease exhibiting both pH and thermostability, which can be used for various applications in industries.

  9. Characterization and phylogenetic analysis of novel polyene type antimicrobial metabolite producing actinomycetes from marine sediments: Bay of Bengal India

    PubMed Central

    Valan, Arasu M; Asha, KRT; Duraipandiyan, V; Ignacimuthu, S; Agastian, P

    2012-01-01

    Objective To isolate and indentify the promising antimicrobial metabolite producing Streptomyces strains from marine sediment samples from Andrapradesh coast of India. Methods Antagonistic actinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1% glucose used as a base for primary screening. Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level. Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method. Results Among the 210 actinomycetes, 64.3% exhibited activity against Gram positive bacteria, 48.5 % showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85 % isolates revealed significant antifungal activity. However, five isolates AP-5, AP-18, AP-41 and AP-70 showed significant antimicrobial activity. The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates. Sequencing analysis indicated that the isolates shared 98.5%-99.8% sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons. The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30°Cfor five days. The antimicrobial activity was assessed using broth micro dilution technique. Each of the culture extracts from these five strains showed a typical polyene-like property. The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Curvularia lunata were 67.5 and 125.0 µg/mL, respectively. Conclusions It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi. PMID:23569851

  10. Antimicrobial Activity and Phylogenetic Analysis of Streptomyces Parvulus Dosmb-D105 Isolated from the Mangrove Sediments of Andaman Islands.

    PubMed

    Baskaran, R; Mohan, P M; Sivakumar, K; Kumar, Ashok

    2016-03-01

    Actinomycetes, especially species of Streptomyces are prolific producers of pharmacologically significant compounds accounting for about 70% of the naturally derived antibiotics that are presently in clinical use. In this study, we used five solvents to extract the secondary metabolites from marine Streptomyces parvulus DOSMB-D105, which was isolated from the mangrove sediments of the South Andaman Islands. Among them, ethyl acetate crude extract showed maximum activity against 11 pathogenic bacteria and six fungi. Presence of bioactive compounds in the ethyl acetate extract was determined using GC-MS and the compounds detected in the ethyl acetate extract were matched with the National Institute of Standards and Technology (NIST) library. Totally eight compounds were identified and the prevalent compounds were 2 steroids, 2 alkaloids, 2 plasticizers, 1 phenolic and 1 alkane. Present study revealed that S. parvulus DOSMB-D105 is a promising species for the isolation of valuable bioactive compounds to combat pathogenic microbes.

  11. The dynamic transcriptional and translational landscape of the model antibiotic producer Streptomyces coelicolor A3(2)

    PubMed Central

    Jeong, Yujin; Kim, Ji-Nu; Kim, Min Woo; Bucca, Giselda; Cho, Suhyung; Yoon, Yeo Joon; Kim, Byung-Gee; Roe, Jung-Hye; Kim, Sun Chang; Smith, Colin P.; Cho, Byung-Kwan

    2016-01-01

    Individual Streptomyces species have the genetic potential to produce a diverse array of natural products of commercial, medical and veterinary interest. However, these products are often not detectable under laboratory culture conditions. To harness their full biosynthetic potential, it is important to develop a detailed understanding of the regulatory networks that orchestrate their metabolism. Here we integrate nucleotide resolution genome-scale measurements of the transcriptome and translatome of Streptomyces coelicolor, the model antibiotic-producing actinomycete. Our systematic study determines 3,570 transcription start sites and identifies 230 small RNAs and a considerable proportion (∼21%) of leaderless mRNAs; this enables deduction of genome-wide promoter architecture. Ribosome profiling reveals that the translation efficiency of secondary metabolic genes is negatively correlated with transcription and that several key antibiotic regulatory genes are translationally induced at transition growth phase. These findings might facilitate the design of new approaches to antibiotic discovery and development. PMID:27251447

  12. Inhibition of Aspergillus parasiticus and cancer cells by marine actinomycete strains

    NASA Astrophysics Data System (ADS)

    Li, Ping; Yan, Peisheng

    2014-12-01

    Ten actinomycete strains isolated from the Yellow Sea off China's coasts were identified as belonging to two genera by 16S rDNA phylogenetic analysis: Streptomyces and Nocardiopsis. Six Streptomyces strains (MA10, 2SHXF01-3, MA35, MA05-2, MA05-2-1 and MA08-1) and one Nocardiopsis strain (MA03) were predicted to have the potential to produce aromatic polyketides based on the analysis of the KSα (ketoacyl-synthase) gene in the type II PKS (polyketides synthase) gene cluster. Four strains (MA03, MA01, MA10 and MA05-2) exhibited significant inhibitory effects on mycelia growth (inhibition rate >50%) and subsequent aflatoxin production (inhibition rate >75%) of the mutant aflatoxigenic Aspergillus parasiticus NFRI-95. The ethyl acetate extracts of the broth of these four strains displayed significant inhibitory effects on mycelia growth, and the IC50 values were calculated (MA03: 0.275 mg mL-1, MA01: 0.106 mg mL-1, MA10: 1.345 mg mL-1 and MA05-2: 1.362 mg mL-1). Five strains (2SHXF01-3, MA03, MA05-2, MA01 and MA08-1) were selected based on their high cytotoxic activities. The ethyl acetate extract of the Nocardiopsis strain MA03 was particularly noted for its high antitumor activity against human carcinomas of the cervix (HeLa), lung (A549), kidney (Caki-1) and liver (HepG2) (IC50: 2.890, 1.981, 3.032 and 2.603 μg mL-1, respectively). The extract also remarkably inhibited colony formation of HeLa cells at an extremely low concentration (0.5 μg mL-1). This study highlights that marine-derived actinomycetes are a huge resource of compounds for the biological control of aflatoxin contamination and the development of novel drugs for human carcinomas.

  13. [Actinomycetes from mangrove and their secondary metabolites].

    PubMed

    Hong, Kui

    2013-11-04

    Mangroves are woody plants located in tropical and subtropical intertidal coastal regions. Driven by the discovery of novel natural products from marine environment, mangrove is becoming a hot spot for actinomycetes resources collection and secondary metabolites (natural products) identification as well as their biosynthesis mechanism investigation. Salinaspora A produced by a Salinispora strain isolated from Bahamas mangrove environment, is in the first clinical trial. Till the time of writing this paper, 24 genera of 11 families and 8 suborders under the actinomycetale have been reported from mangrove, among which 3 are new genera, and 31 are new species. At the same time, secondary metabolites were identified from the mangrove actinomycetes culture, including alkanoids and quinines, azalomycins, antimycins, bezamides and quinazolines, divergolides, indole derivatives, kandenols, macrocyclic dilactones, and the attractive structures, such as the Streptocarbazoles, the multicyclic indolsesquiterpenes, and xiamycin presented unique structures. Their biosynthetic mechanism has also been investigated. Most of the metabolites were isolated from streptomycetes, with a few from Micromonospora and Saccharopolyspora.

  14. Structure of an MmyB-Like Regulator from C. aurantiacus, Member of a New Transcription Factor Family Linked to Antibiotic Metabolism in Actinomycetes

    PubMed Central

    Xu, Qingping; van Wezel, Gilles P.; Chiu, Hsiu-Ju; Jaroszewski, Lukasz; Klock, Heath E.; Knuth, Mark W.; Miller, Mitchell D.; Lesley, Scott A.; Godzik, Adam; Elsliger, Marc-André; Deacon, Ashley M.; Wilson, Ian A.

    2012-01-01

    Actinomycetes are important bacterial sources of antibiotics and other secondary metabolites. Many antibiotic gene clusters are controlled by pathway-specific activators that act in response to growth conditions. Here we present the crystal structure of an MmyB-like transcription regulator MltR (PDB code 3pxp) (Caur_2278) from Chloroflexus aurantiacus, in complex with a fatty acid (myristic acid). MltR is a distant homolog of the methylenomycin activator MmyB and consists of an Xre-type N-terminal DNA-binding domain and a C-terminal ligand-binding module that is related to the Per-Arnt-Sim (PAS) domain. This structure has enabled identification of a new family of bacterial transcription factors that are distributed predominantly in actinomycetes. Bioinformatics analysis of MltR and other characterized family members suggest that they are likely associated with antibiotic and fatty acid metabolism in actinomycetes. Streptomyces coelicolor SCO4944 is a candidate as an ancestral member of the family. Its ortholog in S. griseus, SGR_6891, is induced by A-factor, a γ-butyrolactone that controls antibiotic production and development, and is adjacent to the A-factor synthase gen, afsA. The location of mltR/mmyB homologs, in particular those adjacent to less well-studied antibiotic-related genes, makes them interesting genetic markers for identifying new antibiotic genes. A model for signal-triggered DNA-binding by MltR is proposed. PMID:22844465

  15. Isolation and evaluation of proteolytic actinomycete isolates as novel inducers of pearl millet downy mildew disease protection

    PubMed Central

    Jogaiah, Sudisha; Kurjogi, Mahantesh; Govind, Sharathchandra Ramasandra; Huntrike, Shekar Shetty; Basappa, Vedamurthy Ankala; Tran, Lam-Son Phan

    2016-01-01

    Native endophytic actinomycetes isolated from pearl millet roots were examined for their efficacy to protect pearl millet against downy mildew. Nineteen of 39 isolates were found to be proteolytic, of which 7 strains could directly suppress the sporangium formation of Sclerospora graminicola, the pearl millet downy mildew pathogen. Thus, mycelial suspensions containing either spores or cell-free extract of these 7 isolates were used for seed-coating and -soaking treatments to test for their induction of downy mildew resistance. Results indicated that seed-coating overall provided better protection to downy mildew than seed-soaking. In both treatments, the tested isolates demonstrated differential abilities in downy mildew disease protection, with Streptomyces griseus SJ_UOM-07-09 and Streptosporangium roseum SJ_UOM-18-09 showing the highest protection rates. Additionally, the levels of disease protection conferred by the actinomycetes were just slightly lower than that of the systemic fungicide Apron, suggesting their effectiveness. Further studies revealed that the more rapid root colonization by SJ_UOM-18-09 resulted in faster and higher induced resistance in comparison with SJ_UOM-07-09 under greenhouse conditions, indicating that SJ_UOM-18-09 was superior than SJ_UOM-07-09 in inducing resistance. Results from this study provide comprehensive information on biocontrol functions of SJ_UOM- 18-09 with great potential to control downy mildew disease in pearl millet. PMID:27499196

  16. Isolation, Characterization and Selection of Avermectin-Producing Streptomyces avermitilis Strains From Soil Samples

    PubMed Central

    Siddique, Samia; Syed, Quratulain; Adnan, Ahmad; Qureshi, Fahim Ashraf

    2014-01-01

    Background: Streptomyces avermitilis, belonging to Actinomycetes, is specialized for production of avermectin, used as an anthelmintic and insecticidal agent. It is mostly found in soil and its isolation is very crucial for medically important avermectin production. Objectives: In the present study, 10 bacterial isolates lacking antimicrobial activities were isolated from the soil samples collected from different areas of Lahore, Pakistan. Materials and Methods: Three distinctive localities of Lahore were opted for soil assortment to isolate S. avermitilis. About 50 isolates of Streptomyces species were attained through selective prescreening procedures. All of these isolates were studied for production of the secondary metabolite, avermectin. Different test like soluble pigment color and melanin formation were used for identification. Biochemical characterizations of those isolates closely resembling the control in morphological characteristics, soluble pigment color and melanin formation tests were performed. Results: The 10 selected isolates were identified as the avermectin-producing strain by fermentation and characterized on ISP2 medium for aerial and reverse side mycelia color, soluble pigment color and melanin formation, in comparison with S. avermitilis DSM 41445. The best avermectin-producing isolate S1-C (10.15 mg/L) showed similar result as S. avermitilis D