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1

Glucoamylase production by a newly isolated strain of Aspergillus niger  

SciTech Connect

Glucoamylase production by Aspergillus niger 57 was studied in complex and synthetic media under stationary vs. submerged conditions. Stationary cultivation resulted in significantly greater yields than did submerged culture. Crude enzyme activity was optimum at 60 degrees and pH 4.0.

Sinkar, V.P.; Lewis, N.F.

1982-01-01

2

Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases  

Microsoft Academic Search

In the present study, the extracellular protease activity in a strain of the filamentous fungus Aspergillus niger was investigated and mutant strains deficient in the production of extracellular proteases were isolated. The major protease, which is responsible for 80–85% of the total activity, is aspergillopepsin A, a protein of ca. 43 kDa, the activity of which is inhibited by pepstatin.

Ineke E. Mattern; Johannes M. van Noort; Paul van den Berg; David B. Archer; Ian N. Roberts; Cees A. M. J. J. Hondel

1992-01-01

3

Production of ( R)-1-(4-Bromo-phenyl)-ethanol by locally isolated Aspergillus niger using ram horn peptone  

Microsoft Academic Search

Aspergillus niger EBK-9 was isolated from soil sample. This isolate was evaluated for production of (R)-1-(4-Bromo-phenyl)-ethanol 2 from 1-(4-Bromo-phenyl)-ethanone 1. In this work, the production of the 2 was achieved via fermenter. Glucose, yeast extract and ram horn peptone as medium in fermenter for growth of A. niger was used. A. niger EBK-9 isolate was found to be an effective

Kani Zilbeyaz; Esabi B. Kurbanoglu

2008-01-01

4

Thermal Characterization of Purified Glucose Oxidase from A Newly Isolated Aspergillus Niger UAF-1  

PubMed Central

An intracellular glucose oxidase was isolated from the mycelium extract of a locally isolated strain of Aspergillus niger UAF-1. The enzyme was purified to a yield of 28.43% and specific activity of 135 U mg?1 through ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The enzyme showed high affinity for D-glucose with a Km value of 2.56 mM. The enzyme exhibited optimum catalytic activity at pH 5.5. Temperature optimum for glucose oxidase, catalyzed D-glucose oxidation was 40°C. The enzyme showed a high thermostability having a half-life 30 min, enthalpy of denaturation 99.66 kJ mol?1 and free energy of denaturation 103.63 kJ mol?1. These characteristics suggest the use of glucose oxidase from Aspergillus niger UAF-1 as an analytical reagent and in the design of biosensors for clinical, biochemical and diagnostic assays. PMID:18193107

Anjum Zia, Muhammad; Khalil-ur-Rahman; K. Saeed, Muhammad; Andaleeb, Fozia; I. Rajoka, Muhammad; A. Sheikh, Munir; A. Khan, Iftikhar; I. Khan, Azeem

2007-01-01

5

Naphtho-gamma-pyrone production by Aspergillus niger isolated from stored cottonseed.  

PubMed

Aspergillus niger was found to be the predominant fungal contaminant of stored cottonseed. Seven strains were isolated and grown on rice. The hexane-insoluble material from methylene chloride extracts of 2-week-old cultures contained components toxic to mice. Based on high-pressure thin-layer and liquid chromatographic analyses, the major components in the mixture were eight different naphtho-gamma-pyrones. Of these, the hydrated dimeric naphthopyrones aurasperones B and C occurred in higher yield than aurasperones A, iso-A, and D and the monomeric naphthopyrones flavasperone and rubrofusarin, all of which were present in the mixture. In addition, fonsecin monomethyl ether was isolated. This metabolite may be a precursor in the biosynthesis of the hydrated aurasperones; it has not been identified previously as a metabolite of A. niger. The relative amounts of the different naphthopyrones were dependent on both the growth substrate and the fungal isolate. PMID:6548104

Ehrlich, K C; DeLucca, A J; Ciegler, A

1984-07-01

6

Isolation of epiphytic yeasts with potential for biocontrol of Aspergillus carbonarius and A. niger on grape.  

PubMed

Antagonistic yeasts were isolated from the epiphytic flora associated with grape berries cv. Negroamaro and identified at species level using molecular methods. A total of 144 yeast isolates were tested in a preliminary screening on agar to select isolates showing a killer activity against Aspergillus carbonarius and A. niger, the main species responsible for the accumulation of ochratoxin A in grape. Twenty-eight yeast isolates were selected for their inhibitory effects on the above fungal species and assayed by an in vitro nutritional competition test for their antagonistic capacity towards three selected ochratoxigenic strains. Six yeast isolates belonging to five species, namely 2 isolates of Issatchenkia orientalis and one each of Metschnikowia pulcherrima, Kluyveromyces thermotolerans, Issatchenkia terricola and Candida incommunis, were finally selected and screened on wounded grape berries for their ability to inhibit infection by ochratoxigenic moulds. With the exception of the K. thermotolerans isolate, when inoculated at 10(9) CFU/wound, the other five challenger yeasts reduced the A. carbonarius and A. niger colonization on grape berry (P<0.05). In particular, the best antagonistic activity was shown by the two I. orientalis isolates. Results suggest that antagonist yeasts with the potential to control A. carbonarius and A. niger on grape can be found among the microflora associated with the berries. PMID:16443300

Bleve, Gianluca; Grieco, Francesco; Cozzi, Giuseppe; Logrieco, Antonio; Visconti, Angelo

2006-04-25

7

Isolation and structure of the pectin lyase D-encoding gene from Aspergillus niger.  

PubMed

The filamentous fungus, Aspergillus niger, produces a number of extracellular pectin-degrading enzymes. We present here the isolation and the complete nucleotide sequence of the gene, pelD, coding for a pectin lyase D (PLD), which was previously described as pectin lyase I (Van Houdenhoven, Ph.D. Thesis, Wageningen, 1975). The deduced amino acid (aa) sequence corresponds to 373 aa residues including a signal peptide of 19 aa. The coding region is interrupted by four short introns (57-65 bp). The nucleotide sequence of the 5'- and 3'-flanking regions is also presented and shows no unusual features. By comparing the deduced aa sequences of the A. niger PLD and a number of bacterial pectate lyases, short regions of homology were found despite the different substrate specificities (high methoxyl-pectin versus low methoxyl-pectin or polygalacturonate) of these enzymes. PMID:2373363

Gysler, C; Harmsen, J A; Kester, H C; Visser, J; Heim, J

1990-04-30

8

Thermal Characterization of Purified Glucose Oxidase from A Newly Isolated Aspergillus Niger UAF-1.  

PubMed

An intracellular glucose oxidase was isolated from the mycelium extract of a locally isolated strain of Aspergillus niger UAF-1. The enzyme was purified to a yield of 28.43% and specific activity of 135 U mg(-1) through ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The enzyme showed high affinity for D-glucose with a Km value of 2.56 mM. The enzyme exhibited optimum catalytic activity at pH 5.5. Temperature optimum for glucose oxidase, catalyzed D-glucose oxidation was 40 degrees C. The enzyme showed a high thermostability having a half-life 30 min, enthalpy of denaturation 99.66 kJ mol(-1) and free energy of denaturation 103.63 kJ mol(-1). These characteristics suggest the use of glucose oxidase from Aspergillus niger UAF-1 as an analytical reagent and in the design of biosensors for clinical, biochemical and diagnostic assays. PMID:18193107

Anjum Zia, Muhammad; Khalil-Ur-Rahman; K Saeed, Muhammad; Andaleeb, Fozia; I Rajoka, Muhammad; A Sheikh, Munir; A Khan, Iftikhar; I Khan, Azeem

2007-09-01

9

Ochratoxin A production and amplified fragment length polymorphism analysis of Aspergillus carbonarius, Aspergillus tubingensis, and Aspergillus niger strains isolated from grapes in Italy.  

PubMed

Ochratoxin A is a potent nephrotoxin and a possible human carcinogen that can contaminate various agricultural products, including grapes and wine. The capabilities of species other than Aspergillus carbonarius within Aspergillus section Nigri to produce ochratoxin A from grapes are uncertain, since strain identification is based primarily on morphological traits. We used amplified fragment length polymorphisms (AFLPs) and genomic DNA sequences (rRNA, calmodulin, and beta-tubulin genes) to identify 77 black aspergilli isolated from grape berries collected in a 2-year survey in 16 vineyards throughout Italy. Four main clusters were distinguished, and they shared an AFLP similarity of <25%. Twenty-two of 23 strains of A. carbonarius produced ochratoxin A (6 to 7,500 microg/liter), 5 of 20 strains of A. tubingensis produced ochratoxin A (4 to 130 microg/liter), 3 of 15 strains of A. niger produced ochratoxin A (250 to 360 microg/liter), and none of the 19 strains of Aspergillus "uniseriate" produced ochratoxin A above the level of detection (4 microg/liter). These findings indicate that A. tubingensis is able to produce ochratoxin and that, together with A. carbonarius and A. niger, it may be responsible for the ochratoxin contamination of wine in Italy. PMID:16391107

Perrone, Giancarlo; Mulè, Giuseppina; Susca, Antonia; Battilani, Paola; Pietri, Amedeo; Logrieco, Antonio

2006-01-01

10

Ochratoxin A Production and Amplified Fragment Length Polymorphism Analysis of Aspergillus carbonarius, Aspergillus tubingensis, and Aspergillus niger Strains Isolated from Grapes in Italy  

PubMed Central

Ochratoxin A is a potent nephrotoxin and a possible human carcinogen that can contaminate various agricultural products, including grapes and wine. The capabilities of species other than Aspergillus carbonarius within Aspergillus section Nigri to produce ochratoxin A from grapes are uncertain, since strain identification is based primarily on morphological traits. We used amplified fragment length polymorphisms (AFLPs) and genomic DNA sequences (rRNA, calmodulin, and ?-tubulin genes) to identify 77 black aspergilli isolated from grape berries collected in a 2-year survey in 16 vineyards throughout Italy. Four main clusters were distinguished, and they shared an AFLP similarity of <25%. Twenty-two of 23 strains of A. carbonarius produced ochratoxin A (6 to 7,500 ?g/liter), 5 of 20 strains of A. tubingensis produced ochratoxin A (4 to 130 ?g/liter), 3 of 15 strains of A. niger produced ochratoxin A (250 to 360 ?g/liter), and none of the 19 strains of Aspergillus “uniseriate” produced ochratoxin A above the level of detection (4 ?g/liter). These findings indicate that A. tubingensis is able to produce ochratoxin and that, together with A. carbonarius and A. niger, it may be responsible for the ochratoxin contamination of wine in Italy. PMID:16391107

Perrone, Giancarlo; Mulè, Giuseppina; Susca, Antonia; Battilani, Paola; Pietri, Amedeo; Logrieco, Antonio

2006-01-01

11

Aspergillus niger contains the cryptic phylogenetic species A. awamori.  

PubMed

Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger 'aggregate' represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus acidus, Aspergillus brasiliensis, Aspergillus costaricaensis, Aspergillus lacticoffeatus, Aspergillus piperis, and Aspergillus vadensis. Aspergillus awamori, first described by Nakazawa, has been compared taxonomically with other black aspergilli and recently it has been treated as a synonym of A. niger. Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins ?-tubulin (benA), calmodulin (CaM), and the translation elongation factor-1 alpha (TEF-1?) of a population of A. niger strains isolated from grapes in Europe revealed the presence of a cryptic phylogenetic species within this population, A. awamori. Morphological, physiological, ecological and chemical data overlap occurred between A. niger and the cryptic A. awamori, however the splitting of these two species was also supported by AFLP analysis of the full genome. Isolates in both phylospecies can produce the mycotoxins ochratoxin A and fumonisin B?, and they also share the production of pyranonigrin A, tensidol B, funalenone, malformins, and naphtho-?-pyrones. In addition, sequence analysis of four putative A. awamori strains from Japan, used in the koji industrial fermentation, revealed that none of these strains belong to the A. awamori phylospecies. PMID:22036292

Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena; Varga, János; Frisvad, Jens C; Samson, Robert A

2011-11-01

12

Optimization of the production of thermostable endo-beta-1,4 mannanases from a newly isolated Aspergillus niger gr and Aspergillus flavus gr.  

PubMed

The aim of this work was to establish optimal conditions for the maximum production of endo-beta-1,4 mannanases using cheaper sources. Eight thermotolerant fungal strains were isolated from garden soil and compost samples collected in and around the Gulbarga University campus, India. Two strains were selected based on their ability to produce considerable endo-beta-1,4 mannanases activity while growing in liquid medium at 37 degrees C with locust bean gum (LBG) as the only carbon source. They were identified as Aspergillus niger gr and Aspergillus flavus gr. The experiment to evaluate the effect of different carbon sources, nitrogen sources, temperatures and initial pH of the medium on maximal enzyme production was studied. Enzyme productivity was influenced by the type of polysaccharide used as the carbon source. Copra meal defatted with n-hexane showed to be a better substrate than LBG and guar gum for endo-beta-1,4 mannanases production by A. niger gr (40.011 U/ml), but for A. flavus gr (33.532 U/ml), the difference was not significant. Endo-beta-1,4 mannanases produced from A. niger gr and A. flavus gr have high optimum temperature (65 and 60 degrees C) and good thermostability in the absence of any stabilizers (maintaining 50% of residual activity for 8 and 6 h, respectively, at 60 degrees C) and are stable over in a wide pH range. These new strains offer an attractive alternative source of enzymes for the food and feed processing industries. PMID:18597050

Kote, Naganagouda V; Patil, Aravind Goud G; Mulimani, V H

2009-02-01

13

An electrophoretic karyotype of Aspergillus niger  

Microsoft Academic Search

An electrophoretic karyotype of Aspergillus niger was obtained using contour-clamped homogeneous electric field (CHEF) gel electrophoresis. Chromosomesized DNA was separated into four bands. Seven of the eight linkage groups could be correlated with specific chromosomal bands. For this purpose DNA preparations from seven transformant strains of A. niger each carrying the heterologous amdS gene of Aspergillus nidulans on a different

Alfons J. M. Debets; Edu F. Holub; Klaas Swart; Henk W. J. Broek; Cees J. Bos

1990-01-01

14

Heavy metal biosorption sites in Aspergillus niger  

Microsoft Academic Search

Aspergillus niger is capable of removing heavy metals such as lead, cadmium and copper from aqueous solutions. The role played by various functional groups in the cell wall of A. niger in biosorption of lead, cadmium and copper was investigated. The biomass was subjected to chemical treatments to modify the functional groups, carboxyl, amino and phosphate, to study their role

Anoop Kapoor; T. Viraraghavan

1997-01-01

15

Dye biosorption sites in Aspergillus niger  

Microsoft Academic Search

Aspergillus niger is capable of removing dyes from an aqueous solution. In the study, the roles played by three major functional groups: carboxyl, amino and phosphate, and the lipid fraction in the biomass of A. niger in biosorption of four dyes, Basic Blue 9, Acid Blue 29, Congo Red and Disperse Red 1, were investigated. These functional groups in A.

Yuzhu Fu; T Viraraghavan

2002-01-01

16

Biotransformation of Stypotriol triacetate by Aspergillus niger  

NASA Astrophysics Data System (ADS)

Biological transformation of the meroditerpenoid, stypotriol triacetate ( 1) by the fungi Aspergillus niger, Cunninghamella elegans, Gibberella fujikuroi and Mucor plumbeus was studied. The incubation of 1 with A. niger yielded the new compound 6',14-diacetoxy-stypol-4,5-dione ( 2) whose structure was established by 1H, 13C and 2D NMR and supported by DFT/GIAO.

Areche, Carlos; Vaca, Inmaculada; Labbe, Pamela; Soto-Delgado, Jorge; Astudillo, Luis; Silva, Mario; Rovirosa, Juana; San-Martin, Aurelio

2011-07-01

17

A tyrosinase inhibitor from Aspergillus niger.  

PubMed

Tyrosinase, in the presence of oxygen, is the main culprit in post harvest browning of food products, resulting in the drop in its commercial value. In an effort to seek natural tyrosinase inhibitors for food applications, a screening programme was undertaken. Of the 26 fungal cultures isolated from soil samples of Agumbe forest, India, one isolate S16, identified as Aspergillus niger, gave an inhibition of 84 % against the enzyme. The inhibitor was isolated by following an enzyme inhibition assay guided purification protocol. The structure of the inhibitor was elucidated and found to be kojic acid. The IC50 of the Competitive inhibitor was found to be 8.8 ?g with a Ki of 0.085 mM. PMID:25328242

Vasantha, K Y; Murugesh, C S; Sattur, A P

2014-10-01

18

Phytosterols elevation in bamboo shoot residue through laboratorial scale solid-state fermentation using isolated Aspergillus niger CTBU.  

PubMed

Aspergillus niger CTBU isolated from local decayed bamboo shoot residue was employed to solid-state fermentation (SSF) of bamboo shoot residue to elevate the content of phytosterols. Strain acclimatization was carried out under the fermentation condition using bamboo shoot as substrate for fermentation performance improvement. The optimal fermentation temperature and nitrogen level were investigated using acclimatized strain, and SSF was carried out in a 500-ml Erlenmeyer flask feeding 300-mg bamboo shoot residue chips under the optimal condition (33 °C and feeding 4 % urea), and 1,186 mg (100 g)(-1) of total phytosterol was attained after 5-day fermentation, in comparison, only 523 mg (100 g)(-1) of phytosterol was assayed in fresh shoots residue. HPLC analysis of the main composition of total phytosterols displays that the types of phytosterols and composition ratio of main sterols keep steady. This laboratorial scale SSF unit could be scaled up for raw phytosterols production from discarded bamboo shoot residue and could reduce its cost. PMID:24610040

Zheng, X X; Chen, R S; Shen, Y; Yin, Z Y

2014-04-01

19

Novel metabolites in phenanthrene and pyrene transformation by Aspergillus niger.  

PubMed Central

Aspergillus niger, isolated from hydrocarbon-contaminated soil, was examined for its potential to degrade phenanthrene and pyrene. Two novel metabolites, 1-methoxyphenanthrene and 1-methoxypyrene, were identified by conventional chemical techniques. Minor metabolites identified were 1- and 2-phenanthrol and 1-pyrenol. No 14CO2 evolution was observed in either [14C]phenanthrene or [14C]pyrene cultures. PMID:9212437

Sack, U; Heinze, T M; Deck, J; Cerniglia, C E; Cazau, M C; Fritsche, W

1997-01-01

20

Response surface methodology for optimization of culture conditions for dye decolorization by a fungus, Aspergillus niger HM11 isolated from dye affected soil  

PubMed Central

Background and Objectives Discharge of wastewater from textile dyeing industries has been a problem in terms of pollution and treatment of these waters is a great task. Keeping this in mind, the aim of our current research is to study the effect of various bioprocess variables on decolorization of an azo dye, Congo red, by a fungal isolate, Aspergillus niger HM11. Materials and Methods Central composite design (CCD) and response surface methodology (RSM) have been applied to design experiments to evaluate the interactive effects of the operating variables: on the decolorization of Congo red. A total of 30 experiments were conducted in the present study and a regression coefficient between the variables was generated. Results The RSM indicated that pH 6.0, 150 rpm agitation, incubation time of 36 hrs and a glucose concentration of 1.0% were optimal for maximum decolorization of Congo red and the response indicated excellent evaluation of experimental data. Conclusion From this study, it is very obvious that the fungal isolate, Aspergillus niger HM11 can be used as a promising microbial strain for decolorization of textile dyeing effluent containing similar dyes. PMID:22347575

Karthikeyan, K; Nanthakumar, K; Shanthi, K; Lakshmanaperumalsamy, P

2010-01-01

21

Purification and Characterization of a Dimethoate-Degrading Enzyme of Aspergillus niger ZHY256, Isolated from Sewage  

PubMed Central

A dimethoate-degrading enzyme from Aspergillus niger ZHY256 was purified to homogeneity with a specific activity of 227.6 U/mg of protein. The molecular mass of the purified enzyme was estimated to be 66 kDa by gel filtration and 67 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point was found to be 5.4, and the enzyme activity was optimal at 50°C and pH 7.0. The activity was inhibited by most of the metal ions and reagents, while it was induced by Cu2+. The Michaelis constant (Km) and Vmax for dimethoate were 1.25 mM and 292 ?mol min?1 mg of protein?1, respectively. PMID:11472959

Liu, Yu-Huan; Chung, Ying-Cheng; Xiong, Ya

2001-01-01

22

Biosorption of heavy metals on Aspergillus niger: Effect of pretreatment  

Microsoft Academic Search

The effect of pretreatment of Aspergillus niger biomass on biosorption of lead, cadmium, copper and nickel was studied. Pretreatment of live A. niger biomass using sodium hydroxide, formaldehyde, dimethyl sulphoxide and detergent resulted in significant improvements in biosorption of lead, cadmium and copper in comparison with live A. niger cells. Pretreatment of A. niger reduced biosorption of nickel as compared

A. Kapoor; T. Viraraghavan

1998-01-01

23

Transformation of Aspergillus niger using the homologous orotidine-5?-phosphate-decarboxylase gene  

Microsoft Academic Search

A homologous transformation system for the filamentous fungus Aspergillus niger has been developed, based on the orotidine-5'-phosphate-decarboxylase gene. A. niger Pyr- mutants have been selected from 5-fluoroorotic acid resistant mutants. These mutants were found to comprise two complementation groups, pyrA and pyrB. The A. niger OMP-decarboxylase gene was isolated from a gene library by heterologous hybridization with the Neurospora crassa

Theo Goosen; Gerda Bloemheuvel; Christoph Gysler; Dick A. Bie; Henk W. J. Broek; Klaas Swart

1987-01-01

24

Fingernail Onychomycosis Due to Aspergillus niger.  

PubMed

Onychomycosis is usually caused by dermatophytes, but some species of nondermatophytic molds and yeasts are also associated with nail invasion. Aspergillus niger is a nondermatophytic mold which exists as an opportunistic filamentous fungus in all environments. Here, we report a case of onychomycosis caused by A. niger in a 66-year-old female. The patient presented with a black discoloration and a milky white base and onycholysis on the proximal portion of the right thumb nail. Direct microscopic examination of scrapings after potassium hydroxide (KOH) preparation revealed dichotomous septate hyphae. Repeated cultures on Sabouraud's dextrose agar (SDA) without cycloheximide produced the same black velvety colonies. No colony growth occurred on SDA with cycloheximide slants. Biseriate phialides covering the entire vesicle with radiate conidial heads were observed on the slide culture. The DNA sequence of the internal transcribed spacer region of the clinical sample was a 100% match to that of A. niger strain ATCC 16888 (GenBank accession number AY373852). A. niger was confirmed by KOH mount, colony identification, light microscopic morphology, and DNA sequence analysis. The patient was treated orally with 250 mg terbinafine daily and topical amorolfine 5% nail lacquer for 3 months. As a result, the patient was completely cured clinically and mycologically. PMID:23197914

Kim, Dong Min; Suh, Moo Kyu; Ha, Gyoung Yim; Sohng, Seung Hyun

2012-11-01

25

Chronic bilateral otomycosis caused by Aspergillus niger.  

PubMed

Aspergillus niger, an opportunistic filamentous fungus, was identified as the cause of chronic bilateral otomycosis in a 46-year-old female patient who was unresponsive to different drugs. The patient showed signs of erythema, otalgia, itching, otorrhoea and presence of greyish black coloured mass in both the ear canals. The direct microscopical examination of the ear debris in potassium hydroxide preparations, Giemsa, phase contrast and Gram revealed many thin, branched septate hyphae, condia and conidiophores morphologically indistinguishable from Aspergillus spp. The histopathological section of the ear wax mass by haematoxylin and eosin and periodic acid-Schiff techniques also showed similar fungal elements. The patient responded to 1% solution of mercurochrome. The use of mercurochrome in developing countries like India may be recommended to treat the fungal otitis in patients. We also emphasize that 'Narayan' stain should be routinely employed by microbiology and public health laboratories to study the morphology of pathogenic fungi. PMID:14998406

Mishra, G S; Mehta, Niral; Pal, M

2004-02-01

26

Cloning and expression of a second Aspergillus niger pectin lyase gene ( pel A): Indications of a pectin lyase gene family in A. niger  

Microsoft Academic Search

Using the previously cloned Aspergillus niger N756 pectin lyase D gene as a probe, the corresponding pelD gene has been isolated from a genomic library of the loboratory strain A. niger N400. This gene encodes PLD, previously described as PLI, which is one of the two major pectin lyases isolated from the commeriial pectinase preparation Ultrazym®. Heterologous hybridization of the

J. A. M. Harmsen; M. A. Kusters-van Someren; J. Visser

1990-01-01

27

Fumonisin and Ochratoxin Production in Industrial Aspergillus niger Strains  

PubMed Central

Aspergillus niger is perhaps the most important fungus used in biotechnology, and is also one of the most commonly encountered fungi contaminating foods and feedstuffs, and occurring in soil and indoor environments. Many of its industrial applications have been given GRAS status (generally regarded as safe). However, A. niger has the potential to produce two groups of potentially carcinogenic mycotoxins: fumonisins and ochratoxins. In this study all available industrial and many non-industrial strains of A. niger (180 strains) as well as 228 strains from 17 related black Aspergillus species were examined for mycotoxin production. None of the related 17 species of black Aspergilli produced fumonisins. Fumonisins (B2, B4, and B6) were detected in 81% of A. niger, and ochratoxin A in 17%, while 10% of the strains produced both mycotoxins. Among the industrial strains the same ratios were 83%, 33% and 26% respectively. Some of the most frequently used strains in industry NRRL 337, 3112 and 3122 produced both toxins and several strains used for citric acid production were among the best producers of fumonisins in pure agar culture. Most strains used for other biotechnological processes also produced fumonisins. Strains optimized through random mutagenesis usually maintained their mycotoxin production capability. Toxigenic strains were also able to produce the toxins on media suggested for citric acid production with most of the toxins found in the biomass, thereby questioning the use of the remaining biomass as animal feed. In conclusion it is recommended to use strains of A. niger with inactive or inactivated gene clusters for fumonisins and ochratoxins, or to choose isolates for biotechnological uses in related non-toxigenic species such as A. tubingensis, A. brasiliensis, A vadensis or A. acidus, which neither produce fumonisins nor ochratoxins. PMID:21853139

Frisvad, Jens C.; Larsen, Thomas O.; Thrane, Ulf; Meijer, Martin; Varga, Janos; Samson, Robert A.; Nielsen, Kristian F.

2011-01-01

28

Characterization of two forms of glucoamylase from aspergillus niger  

Microsoft Academic Search

Aspergillus niger glucoamylases GI and GII (E.C. 3.2.1.3) were isolated from a commercial enzyme preparation by ammonium sulfate\\u000a precipitation followed by DEAE-cellulose ion exchange chromatography. Both enzymes consist of a single glycosylated polypeptide\\u000a chain. The molecular weights of GI and GII were determined by sedimentation equilibrium ultracentrifugation to 52,000 and\\u000a 46,000, respectively, and by molecular sieving to 65,000 and 55,000.

Birte Svensson; Torben Graves Svendsen; IB Svendsen; Takuo Sakai; Martin Ottesen

1982-01-01

29

Structure of the Aspergillus niger pel A gene and its expression in Aspergillus niger and Aspergillus nidulans  

Microsoft Academic Search

Aspergillus niger pectin lyases are encoded by a multigene family. The complete nucleotide sequence of the pectin lyase PLA-encoding gene pelA has been determined. Comparison of the deduced amino acid sequence with the deduced amino acid sequence of the other characterized pectin lyase, PLD, shows that the proteins share 69% amino acid identity. When grown on media with pectin as

Margo A. Kusters-van Someren; Jan A. M. Harmsen; Harry C. M. Kester; Jaap Visser

1991-01-01

30

Putative Aspergillus niger-induced oxalate nephrosis in sheep.  

PubMed

A sheep farmer provided a maize-based brewer's grain (mieliemaroek) and bales of Eragrostis curvula hay to ewes and their lambs, kept on zero-grazing in pens. The 'mieliemaroek' was visibly mouldy. After 14 days in the feedlot, clinical signs, including generalised weakness, ataxia of the hind limbs, tremors and recumbency, were noticed. Six ewes died within a period of 7 days. A post mortem examination was performed on 1 ewe. The carcass appeared to be cachectic with mild effusions into the body cavities; mild lung congestion and pallor of the kidneys were observed. Microscopical evaluation revealed nephrosis and birefringent oxalate crystals in the renal tubules when viewed under polarised light. A provisional diagnosis of oxalate nephrosis with subsequent kidney failure was made. Amongst other fungi, Aspergillus niger was isolated from 'mieliemaroek' samples submitted for fungal culture and identification. As A. niger is known to synthesise oxalates, a qualitative screen to detect oxalic acid in the mieliemaroek and purified A. niger isolates was performed using high-performance liquid chromatography (HPLC). Oxalic acid was detected, which supported a diagnosis of soluble oxalate-induced nephropathy. PMID:19653520

Botha, C J; Truter, M; Bredell, T; Lange, L; Mülders, M S G

2009-03-01

31

On the safety of Aspergillus niger – a review  

Microsoft Academic Search

.   \\u000a Aspergillus niger is one of the most important microorganisms used in biotechnology. It has been in use already for many decades to produce\\u000a extracellular (food) enzymes and citric acid. In fact, citric acid and many A. niger enzymes are considered GRAS by the United States Food and Drug Administration. In addition, A. niger is used for biotransformations and waste

E. Schuster; N. Dunn-Coleman; J. Frisvad; P. van Dijck

2002-01-01

32

Invasive Aspergillus niger complex infections in a Belgian tertiary care hospital.  

PubMed

The incidence of invasive infections caused by the Aspergillus niger species complex was 0.043 cases/10 000 patient-days in a Belgian university hospital (2005-2011). Molecular typing was performed on six available A. niger complex isolates involved in invasive disease from 2010 to 2011, revealing A. tubingensis, which has higher triazole minimal inhibitory concentrations, in five out of six cases. PMID:24102876

Vermeulen, E; Maertens, J; Meersseman, P; Saegeman, V; Dupont, L; Lagrou, K

2014-05-01

33

Antifungal activity of strains of lactic acid bacteria isolated from a semolina ecosystem against Penicillium roqueforti, Aspergillus niger and Endomyces fibuliger contaminating bakery products.  

PubMed

Thirty samples of Italian durum wheat semolina and whole durum wheat semolina, generally used for the production of Southern Italy's traditional breads, were subjected to microbiological analysis in order to explore their lactic acid bacteria (LAB) diversity and to find strains with antifungal activity. A total of 125 presumptive LAB isolates (Gram-positive and catalase-negative) were characterized by repetitive extragenic palindromic-PCR (REP-PCR) and sequence analysis of the 16S rRNA gene, leading to the identification of the following species: Weissella confusa, Weissella cibaria, Leuconostoc citreum, Leuconostoc mesenteroides, Lactococcus lactis, Lactobacillus rossiae and Lactobacillus plantarum. The REP-PCR results delineated 17 different patterns whose cluster analysis clearly differentiated W. cibaria from W. confusa isolates. Seventeen strains, each characterized by a different REP-PCR pattern, were screened for their antifungal properties. They were grown in a flour-based medium, comparable to a real food system, and the resulting fermentation products (FPs) were tested against fungal species generally contaminating bakery products, Aspergillus niger, Penicillium roqueforti and Endomyces fibuliger. The results of the study indicated a strong inhibitory activity - comparable to that obtained with the common preservative calcium propionate (0.3% w/v) - of ten LAB strains against the most widespread contaminant of bakery products, P. roqueforti. The screening also highlighted the unexplored antifungal activity of L. citreum, L. rossiae and W. cibaria (1 strain), which inhibited all fungal strains to the same or a higher extent compared with calcium propionate. The fermentation products of these three strains were characterized by low pH values, and a high content of lactic and acetic acids. PMID:19243908

Valerio, Francesca; Favilla, Mara; De Bellis, Palmira; Sisto, Angelo; de Candia, Silvia; Lavermicocca, Paola

2009-09-01

34

21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.  

Code of Federal Regulations, 2011 CFR

...PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173...Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger... from the carbohydrase and cellulase enzyme product. (d) The additive is...

2011-04-01

35

21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.  

Code of Federal Regulations, 2010 CFR

...PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173...Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger... from the carbohydrase and cellulase enzyme product. (d) The additive is...

2010-04-01

36

21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.  

Code of Federal Regulations, 2013 CFR

...PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173...Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger... from the carbohydrase and cellulase enzyme product. (d) The additive is...

2013-04-01

37

21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.  

Code of Federal Regulations, 2014 CFR

...PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173...Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger... from the carbohydrase and cellulase enzyme product. (d) The additive is...

2014-04-01

38

21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.  

Code of Federal Regulations, 2012 CFR

...PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173...Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger... from the carbohydrase and cellulase enzyme product. (d) The additive is...

2012-04-01

39

Characterization of the fumonisin B2 biosynthetic gene cluster in Aspergillus niger and A. awamori.  

Technology Transfer Automated Retrieval System (TEKTRAN)

Aspergillus niger and A. awamori strains isolated from grapes cultivated in Mediterranean basin were examined for fumonisin B2 (FB2) production and presence/absence of sequences within the fumonisin biosynthetic gene (fum) cluster. Presence of 13 regions in the fum cluster was evaluated by PCR assay...

40

Conversion of fusaric acid to fusarinol by Aspergillus niger: A detoxification reaction  

Technology Transfer Automated Retrieval System (TEKTRAN)

The fungus Fusarium oxysporum causes wilt diseases of plants and produces a potent phytotoxin fusaric acid (FA) which is also toxic to many microorganisms. An Aspergillus strain with high tolerance to FA was isolated from soil. HPLC analysis of culture filtrates from A. niger grown with the addition...

41

Identification and characterization of a second polygalacturonase gene of Aspergillus niger  

Microsoft Academic Search

The filamentous fungus Aspergillus niger produces several endopolygalacturonases that are involved in the degradation of pectin. PGI, the enzyme representing the second most abundant activity in a commmercial enzyme preparation, was further characterized and the corresponding gene was isolated. The nucleotide sequence of the pgaI gene was determined and the protein coding region was found to be interrupted by two

H. J. D. Bussink; K. B. Brouwer; L. H. Graaff; H. C. M. Kester; J. Visser

1991-01-01

42

Aspergillus Niger Genomics: Past, Present and into the Future  

SciTech Connect

Aspergillus niger is a filamentous ascomycete fungus that is ubiquitous in the environment and has been implicated in opportunistic infections of humans. In addition to its role as an opportunistic human pathogen, A. niger is economically important as a fermentation organism used for the production of citric acid. Industrial citric acid production by A. niger represents one of the most efficient, highest yield bioprocesses in use currently by industry. The genome size of A. niger is estimated to be between 35.5 and 38.5 megabases (Mb) divided among eight chromosomes/linkage groups that vary in size from 3.5 - 6.6 Mb. Currently, there are three independent A. niger genome projects, an indication of the economic importance of this organism. The rich amount of data resulting from these multiple A. niger genome sequences will be used for basic and applied research programs applicable to fermentation process development, morphology and pathogenicity.

Baker, Scott E.

2006-09-01

43

A new diketopiperazine heterodimer from an endophytic fungus Aspergillus niger.  

PubMed

One new diketopiperazine heterodimer, asperazine A (1), and eight known compounds, asperazine (2), cyclo(d-Phe-l-Trp) (3), cyclo(l-Trp-l-Trp) (4), 4-(hydroxymethyl)-5,6-dihydro-pyran-2-one (5), walterolactone A (6), and campyrones A-C (7-9), were isolated from an endophytic fungus Aspergillus niger. Their structures were determined unequivocally on the basis of extensive spectroscopic data analysis. This is the first report of the presence of compound 3 as a natural product. Cytotoxicity test against human cancer cell lines PC3, A2780, K562, MBA-MD-231, and NCI-H1688 revealed that compounds 1 and 2 had weak activities. PMID:25401948

Li, Xiao-Bin; Li, Yue-Lan; Zhou, Jin-Chuan; Yuan, Hui-Qing; Wang, Xiao-Ning; Lou, Hong-Xiang

2015-02-01

44

Removal of heavy metals using the fungus Aspergillus niger  

Microsoft Academic Search

There is a need to develop technologies that can remove toxic heavy metal ions found in wastewaters. Microorganisms are known to remove heavy metal ions from water. In this study the potential of the fungus Aspergillus niger to remove lead, cadmium, copper and nickel ions was evaluated. A. niger biomass pretreated by boiling in 0.1N NaOH solution for 15 min

Anoop Kapoor; T Viraraghavan; D. Roy Cullimore

1999-01-01

45

Production of citric acid with immobilized Aspergillus niger  

Microsoft Academic Search

The spores of Aspergillus niger were entrapped in calcium-alginate beads and precultivated in growth media with various amounts of nitrogen. During the following citric acid production in shaking cultures an optimum of acid formation and yield was observed after the precultivation with 100–200 mg\\/l NH4NO3. The productivity of the immobilized Aspergillus was found to be 1.5 times higher than in

H. Eikmeier; H. J. Rehm

1984-01-01

46

Screening a Strain of Aspergillus niger and Optimization of Fermentation Conditions for Degradation of Aflatoxin B1 †  

PubMed Central

Aflatoxin B1, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1) was able to degrade aflatoxin B1 after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B1 after 48 h of fermentation in nutrient broth (NB). Optimization of fermentation conditions for aflatoxin B1 degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B1 was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B1 degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B1 degradation by the supernatant were examined. Results indicated that aflatoxin B1 degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment. PMID:25401962

Zhang, Wei; Xue, Beibei; Li, Mengmeng; Mu, Yang; Chen, Zhihui; Li, Jianping; Shan, Anshan

2014-01-01

47

Hydrolase production by Aspergillus niger in solid-state cultivation  

Microsoft Academic Search

A production of macerating enzymes which liquefy and hydrolyze the mandarin orange peel was studied in a solid state cultivation of Aspergillus niger on wheat bran substrate. Solid state cultivation in a 2 l drum fermenter capable of interchangeable operation under dynamic or static conditions were carried out maintaining the moisture content of the substrate at 32, 39, 46, 56,

Naomichi Nishio; Kiyoshi Tai; Shiro Nagai

1979-01-01

48

Stable accumulation of Aspergillus niger phytase in transgenic tobacco leaves  

Microsoft Academic Search

Phytase from Aspergillus niger increases the availability of phos- phorus from feed for monogastric animals by releasing phosphate from the substrate phytic acid. A phytase cDNA was constitutively expressed in transgenic tobacco (Nicofiana tabacum) plants. Secre- tion of the protein to the extracellular fluid was established by use of the signal sequence from the tobacco pathogen-related protein S. lhe specific

Theo C. Verwoerd; Paridon van P. A; Oosen van A. J. J; Lent van J. W. M; André Hoekema; Jan Pen

1995-01-01

49

Function of Conserved Tryptophans in the Aspergillus niger Glucoamylase 1 Starch Binding Domain  

E-print Network

Function of Conserved Tryptophans in the Aspergillus niger Glucoamylase 1 Starch Binding Domain of tryptophan residues in the granular starch binding domain (SBD) of glucoamylase 1 from Aspergillus niger. Wild-type SBD and three variant (W563K, W590K, and W615K) proteins were produced using an A. niger

Williamson, Mike P.

50

Biochemical Engineering Journal 8 (2001) 187193 Enhanced heterologous protein production in Aspergillus niger through  

E-print Network

in Aspergillus niger through pH control of extracellular protease activity Dara O'Donnella, Liping Wanga 2000; accepted 15 February 2001 Abstract The extracellular protease activity of Aspergillus niger AB4 glucose in the culture medium had been completely utilized. When grown at pH 6, A. niger protease activity

Gu, Tingyue

51

Solution structure of the granular starch binding domain of Aspergillus niger glucoamylase bound to -cyclodextrin  

E-print Network

Solution structure of the granular starch binding domain of Aspergillus niger glucoamylase bound the catalytic domains of hydrolytic enzymes. Glucoamylase 1 (G1) from Aspergillus niger, an enzyme used widely of insoluble polysaccharides. Results: The solution structure of the SBD of A. niger G1 bound to -cyclodextrin

Williamson, Mike P.

52

Production of extremophilic bacterial cellulase enzymes in aspergillus niger.  

SciTech Connect

Enzymes can be used to catalyze a myriad of chemical reactions and are a cornerstone in the biotechnology industry. Enzymes have a wide range of uses, ranging from medicine with the production of pharmaceuticals to energy were they are applied to biofuel production. However, it is difficult to produce large quantities of enzymes, especially if they are non-native to the production host. Fortunately, filamentous fungi, such as Aspergillus niger, are broadly used in industry and show great potential for use a heterologous enzyme production hosts. Here, we present work outlining an effort to engineer A. niger to produce thermophilic bacterial cellulases relevant to lignocellulosic biofuel production.

Gladden, John Michael

2013-09-01

53

Production of inulinase using tap roots of dandelion ( Taraxacum officinale) by Aspergillus niger  

Microsoft Academic Search

Various inulin containing vegetal substrates were evaluated for inulinase production by an indigenous isolate, Aspergillus niger NK-126. Highest inulinase activity was observed with dandelion tap root extract (52.3IU\\/ml). The enzyme activity was fourfold higher than that observed in media containing pure chicory inulin (12.3IU\\/ml). The fungus showed good growth on a medium containing 40% (v\\/v) of dandelion tap root extract

Naveen Kango

2008-01-01

54

Production of cellulase and xylanase in a bubble gum column using immobilized Aspergillus niger KKS  

SciTech Connect

Aspergillus niger KKS, isolated from a farmland near Suwon, was immobilized on Celite and polyurethane foams. Enzyme activities produced by the immobilized cell system in a bubble column were higher than that of shake-flask culture. The enzyme productivities were twice as high. {Beta}-Glucosidase, {Beta}-xylosidase, and xylanase activities obtained in a bubble column were significant when the ground rice straw was used as a substrate. 9 refs., 2 figs., 3 tabs.

Kang, Seong-Woo; Kim, Seung-Woo [Univ. of Suwon (Korea, Republic of); Lee, Jin-Suk [Korea Institute of Energy Research, Daejeon (Korea, Republic of)

1995-05-01

55

Expression and sequence comparison of the Aspergillus niger and Aspergillus tubigensis genes encoding polygalacturonase II  

Microsoft Academic Search

The structure and expression of the polygalacturonase-encoding pgaII genes of two recently recognized species, Aspergillus niger and Aspergillus tubigensis, was investigated. While the structure of the pgaII genes is very similar, showing 83% DNA sequence identity and 94% identity at the amino acid level, they have diverged significantly. The NH2-terminal sequence suggests that these PGs are made as pre pro-proteins

Hendrik J. D. Bussink; Frank P. Buxton; Jaap Visser

1991-01-01

56

Cloning and expression of a second Aspergillus niger pectin lyase gene (pelA): indications of a pectin lyase gene family in A. niger.  

PubMed

Using the previously cloned Aspergillus niger N756 pectin lyase D gene as a probe, the corresponding pelD gene has been isolated from a genomic library of the laboratory strain A. niger N400. This gene encodes PLD, previously described as PLI, which is one of the two major pectin lyases isolated from the commercial pectinase preparation Ultrazym. Heterologous hybridization of the A. niger N400 genomic library with the pelD gene led to the isolation of another five genes: pelA, B, C, E, and F. These genes differ in their hybridization patterns with probes containing either the entire pelD gene, or 5' or 3' parts thereof. By partial sequencing, and expression in an A. niger transformant containing multiple copies of the pelA gene, we show that this gene, which hybridizes strongest with the pelD gene, encodes the other major pectin lyase from Ultrazym, PLII. PMID:2225145

Harmsen, J A; Kusters-van Someren, M A; Visser, J

1990-08-01

57

Fractionation of ?-Glucosidases and Related Extracellular Enzymes from Aspergillus niger  

PubMed Central

Industrial concentrates from Aspergillus niger culture filtrates were fractionated by ion-exchange and adsorption chromatography. Several other types of hydrolases were completely removed. Eight partially purified components were obtained. Using specific activity as an estimate of purification, one aryl-?-glucosidase was purified 35-fold. Another component showed 147-fold purification using a viscosimetric assay with carboxymethylcellulose as substrate. The aryl-?-glucosidase was distinctly more thermolabile than the carboxymethylcellulase. PMID:13930396

Li, L.-h.; King, K. W.

1963-01-01

58

Aspergillus niger mutants with increased glucose oxidase production  

Microsoft Academic Search

Aspergillus niger NRRL-3, an organism used for the industrial scale production of d-gluconic acid and glucose oxidase (EC 1.1.3.4), was subjected to mutagenesis and selection for acid production on diagnostic media containing methyl red. The plates contained 0.1 M d-glucose, a concentration that does not produce a color change in the medium surrounding mycelia of the parental strain under the

John Markwell; Laura G. Frakes; Eugene C. Brott; John Osterman; Fred W. Wagner

1989-01-01

59

Purification and immobilization of Aspergillus niger. beta. -xylosidase  

SciTech Connect

..beta..-Xylosidase from a commercial Aspergillus niger preparation was purified by differential ammonium sulfate precipitation and either gel permeation or cation exchange chromatography, giving 16-fold purification in 32% yield for the first technique or 27-fold purification in 19% yield for the second. Enzyme prepared by this method was immobilized to 10 different carriers, but only when it was bound to alumina with TiCl/sub 4/ and to alkylamine porous silica with glutaraldehyde were substantial efficiencies and stabilities achieved.

Oguntimein, G.B.; Reilly, P.J.

1980-01-01

60

Regulation of the glaA gene of Aspergillus niger  

Microsoft Academic Search

The glucoamylase gene of Aspergillus niger, glaA, is expressed at high levels in the presence of starch. We have determined the nucleotide sequence of 1966 bp of the 5' flanking region of the glaA gene and have begun to identify sequences important for the control of glaA expression by deletion analysis. Constructs containing deletions extending into the glaA gene promotor

Timothy Fowler; Randy M. Berka; Michael Ward

1990-01-01

61

Analytical and computational approaches to define the Aspergillus niger secretome  

SciTech Connect

We used computational and mass spectrometric approaches to characterize the Aspergillus niger secretome. The 11,200 gene models predicted in the genome of A. niger strain ATCC 1015 were the data source for the analysis. Depending on the computational methods used, 691 to 881 proteins were predicted to be secreted proteins. We cultured A. niger in six different media and analyzed the extracellular proteins produced using mass spectrometry. A total of 222 proteins were identified, with 39 proteins expressed under all six conditions and 74 proteins expressed under only one condition. The secreted proteins identified by mass spectrometry were used to guide the correction of about 20 gene models. Additional analysis focused on extracellular enzymes of interest for biomass processing. Of the 63 glycoside hydrolases predicted to be capable of hydrolyzing cellulose, hemicellulose or pectin, 94% of the exo-acting enzymes and only 18% of the endo-acting enzymes were experimentally detected.

Tsang, Adrian; Butler, Gregory D.; Powlowski, Justin; Panisko, Ellen A.; Baker, Scott E.

2009-03-01

62

Expression of the Aspergillus terreus itaconic acid biosynthesis cluster in Aspergillus niger  

PubMed Central

Background Aspergillus terreus is a natural producer of itaconic acid and is currently used to produce itaconic acid on an industrial scale. The metabolic process for itaconic acid biosynthesis is very similar to the production of citric acid in Aspergillus niger. However, a key enzyme in A. niger, cis-aconitate decarboxylase, is missing. The introduction of the A. terreus cadA gene in A. niger exploits the high level of citric acid production (over 200 g per liter) and theoretically can lead to production levels of over 135 g per liter of itaconic acid in A. niger. Given the potential for higher production levels in A. niger, production of itaconic acid in this host was investigated. Results Expression of Aspergillus terreus cis-aconitate decarboxylase in Aspergillus niger resulted in the production of a low concentration (0.05 g/L) of itaconic acid. Overexpression of codon-optimized genes for cis-aconitate decarboxylase, a mitochondrial transporter and a plasma membrane transporter in an oxaloacetate hydrolase and glucose oxidase deficient A. niger strain led to highly increased yields and itaconic acid production titers. At these higher production titers, the effect of the mitochondrial and plasma membrane transporters was much more pronounced, with levels being 5–8 times higher than previously described. Conclusions Itaconic acid can be produced in A. niger by the introduction of the A. terreus cis-aconitate decarboxylase encoding cadA gene. This results in a low itaconic acid production level, which can be increased by codon-optimization of the cadA gene for A. niger. A second crucial requirement for efficient production of itaconic acid is the expression of the A. terreus mttA gene, encoding a putative mitochondrial transporter. Expression of this transporter results in a twenty-fold increase in the secretion of itaconic acid. Expression of the A. terreus itaconic acid cluster consisting of the cadA gene, the mttA gene and the mfsA gene results in A. niger strains that produce over twenty five-fold higher levels of itaconic acid and show a twenty-fold increase in yield compared to a strain expressing only CadA. PMID:24438100

2014-01-01

63

Fumonisin B(2) production by Aspergillus niger from grapes and natural occurrence in must.  

PubMed

Aspergillus niger has been recently found to produce fumonisin B(2) (FB(2)). Thirty-one strains belonging to four Aspergillus species isolated from grape were evaluated for FB(2) production on agar plates. Four out of eight strains of A. niger produced FB(2) (29-293 microg g(-1)). None of the strains of A. uvarum (n = 7), A. tubingensis (8) and A. carbonarius (8) produced detectable amounts of toxin. The capability to produce FB(2) was also confirmed by some A. niger strains artificially inoculated on grape berries. Natural occurrence of FB(2), at levels of 0.01 and 0.4 microg ml(-1), was found in two samples of must collected in Apulian cellars in 2007. This is the first report of FB(2) contamination in must. These findings suggest that there is a potential risk of exposure to FB(2) in the grape-wine chain for consumers and that A. niger may represent the major fumonisin-producing species among black Aspergilli occurring on grapes. PMID:19742356

Logrieco, A; Ferracane, R; Haidukowsky, M; Cozzi, G; Visconti, A; Ritieni, A

2009-11-01

64

Removal of cadmium, chromium, lead and copper from urban sewage by Aspergillus Niger and Pseudomonas fluorescens  

Microsoft Academic Search

Aspergillus Niger and Pseudomonas fluorescens have the ability to absorb heavy metals during the stationary phase of growth. To improve the removal the efficiency of the oxidation ditch, we have introduced Aspergillus Niger and Pseudomonas fluorescens to the traditional Carrousel oxidation ditch process and the comparative experimental study were carried out when the Hydraulic Retention Time (HRT) was 36h. According

Haihua Li; Guoqiang Bai; Yingying Fu; Yanyan Jin

2011-01-01

65

Influence of manganese on morphology and cell wall composition of Aspergillus niger during citric acid fermentation  

Microsoft Academic Search

Morphology and cell wall composition of Aspergillus niger were studied under conditions of manganese sufficient or deficient cultivation in an otherwise citric acid producing medium. Omission of Mn2+ (less than 10-7 M) from the nutrient medium of Aspergillus niger results in abnormal morphological development which is characterized by increased spore swelling, and squat, bulbeous hyphae. Fractionation and analysis of manganese

Monika Kisser; C. P. Kubicek; M. Röhr

1980-01-01

66

Samenvatting Aspergillus niger is a filamenteuze schimmel die wereldwijd verspreid is en veelvuldig  

E-print Network

Samenvatting Aspergillus niger is a filamenteuze schimmel die wereldwijd verspreid is en veelvuldig voorkomt op rottend planten materiaal. Als een saprophyt speelt A. niger een, scheidt A. niger een scala aan hydrolytische enzymen uit. Veel van de extracellulaire enzymen van A

Hille, Sander

67

Bioremediation of CCA-C treated wood by Aspergillus niger fermentation  

Microsoft Academic Search

This study evaluated the potential of the fungus Aspergillus niger to remove copper, chromium, and arsenic from waste wood treated with chromated copper arsenate (CCA) wood preservative. The removal of heavy metals by A. niger was carried out in two stages. In the first stage, A. niger was cultivated in carbohydrates media in order to produce large quantities of oxalic

S. N. Kartal; T. Kakitani; Y. Imamura

2004-01-01

68

Salmonella Biofilm Formation on Aspergillus niger Involves Cellulose – Chitin Interactions  

PubMed Central

Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to and forms biofilms on the hyphae of the common fungus, Aspergillus niger. Several Salmonella enterica serovars displayed a similar interaction, whereas other bacterial species were unable to bind to the fungus. Bacterial attachment to chitin, a major constituent of fungal cell walls, mirrored this specificity. Pre-incubation of S. Typhimurium with N-acetylglucosamine, the monomeric component of chitin, reduced binding to chitin beads by as much as 727-fold and inhibited attachment to A. niger hyphae considerably. A cellulose-deficient mutant of S. Typhimurium failed to attach to chitin beads and to the fungus. Complementation of this mutant with the cellulose operon restored binding to chitin beads to 79% of that of the parental strain and allowed for attachment and biofilm formation on A. niger, indicating that cellulose is involved in bacterial attachment to the fungus via the chitin component of its cell wall. In contrast to cellulose, S. Typhimurium curli fimbriae were not required for attachment and biofilm development on the hyphae but were critical for its stability. Our results suggest that cellulose–chitin interactions are required for the production of mixed Salmonella-A. niger biofilms, and support the hypothesis that encounters with chitinaceous alternate hosts may contribute to the ecological success of human pathogens. PMID:22003399

Brandl, Maria T.; Carter, Michelle Q.; Parker, Craig T.; Chapman, Matthew R.; Huynh, Steven; Zhou, Yaguang

2011-01-01

69

FluG affects secretion in colonies of Aspergillus niger.  

PubMed

Colonies of Aspergillus niger are characterized by zonal heterogeneity in growth, sporulation, gene expression and secretion. For instance, the glucoamylase gene glaA is more highly expressed at the periphery of colonies when compared to the center. As a consequence, its encoded protein GlaA is mainly secreted at the outer part of the colony. Here, multiple copies of amyR were introduced in A. niger. Most transformants over-expressing this regulatory gene of amylolytic genes still displayed heterogeneous glaA expression and GlaA secretion. However, heterogeneity was abolished in transformant UU-A001.13 by expressing glaA and secreting GlaA throughout the mycelium. Sequencing the genome of UU-A001.13 revealed that transformation had been accompanied by deletion of part of the fluG gene and disrupting its 3' end by integration of a transformation vector. Inactivation of fluG in the wild-type background of A. niger also resulted in breakdown of starch under the whole colony. Asexual development of the ?fluG strain was not affected, unlike what was previously shown in Aspergillus nidulans. Genes encoding proteins with a signal sequence for secretion, including part of the amylolytic genes, were more often downregulated in the central zone of maltose-grown ?fluG colonies and upregulated in the intermediate part and periphery when compared to the wild-type. Together, these data indicate that FluG of A. niger is a repressor of secretion. PMID:25370014

Wang, Fengfeng; Krijgsheld, Pauline; Hulsman, Marc; de Bekker, Charissa; Müller, Wally H; Reinders, Marcel; de Vries, Ronald P; Wösten, Han A B

2015-01-01

70

Steady-state shear characteristics of Aspergillus niger broths  

SciTech Connect

It can be difficult to obtain reliable rheological data for filamentous fermentation broths using conventional instruments. One common approach is to measure the torque drawn by an impeller rotating in the suspension. Many previous workers have assumed that the applicable shear rate in such a device is related to the impeller speed by a fluid-independent constant determined by calibration with Newtonian and non-Newtonian fluids. The rheology of Aspergillus niger broths have been characterized using the impeller viscometer approach. The changes in the broth rheology were measured, and used to interpret the growth of biomass and the evolution of the microorganism morphology.

Svihla, C.K.; Dronawat, S.N.; Hanley, T.R. [Univ. of Louisville, KY (United States)

1995-12-31

71

Cloning and Expression of Gumboro VP2 Antigen in Aspergillus niger  

PubMed Central

Background Infectious Bursal Disease Virus (IBDV) causes a highly immunosuppressive disease in chickens and is a pathogen of major economic importance to the poultry industry worldwide. The VP2 protein is the major host-protective immunogen of IBDV and has been considered as a potential subunit vaccine against the disease. VP2 coding sequence was cloned in an inducible fungal vector and the protein was expressed in Aspergillus niger (A. niger). Methods Aiming at a high level of expression, a multicopy AMA1-pyrG-based episomal construct driven by a strong inducible promoter, glaA, was prepared and used in transformation of A. niger pyrG-protoplasts. SDS-PAGE and western blot analysis was carried out to confirm the expression of the protein. Results A number of pyrG + positive transformants were isolated and the presence of expression cassette was confirmed. Western blot analysis of one of these recombinant strains using monospecific anti-VP2 antibodies demonstrated the successful expression of the protein. The recombinant protein was also detected by serum obtained from immunized chicken. Conclusion In the present study, we have generated a recombinant A. niger strain expressing VP2 protein intracellulary. This recombinant strain of A. niger may have potential applications in oral vaccination against IBDV in poultry industry. PMID:23626875

Azizi, Mohammad; Yakhchali, Bagher; Ghamarian, Abdolreza; Enayati, Somayeh; Khodabandeh, Mahvash; Khalaj, Vahid

2013-01-01

72

Cellulase Production from Spent Lignocellulose Hydrolysates by Recombinant Aspergillus niger?  

PubMed Central

A recombinant Aspergillus niger strain expressing the Hypocrea jecorina endoglucanase Cel7B was grown on spent hydrolysates (stillage) from sugarcane bagasse and spruce wood. The spent hydrolysates served as excellent growth media for the Cel7B-producing strain, A. niger D15[egI], which displayed higher endoglucanase activities in the spent hydrolysates than in standard medium with a comparable monosaccharide content (e.g., 2,100 nkat/ml in spent bagasse hydrolysate compared to 480 nkat/ml in standard glucose-based medium). In addition, A. niger D15[egI] was also able to consume or convert other lignocellulose-derived compounds, such as acetic acid, furan aldehydes, and phenolic compounds, which are recognized as inhibitors of yeast during ethanolic fermentation. The results indicate that enzymes can be produced from the stillage stream as a high-value coproduct in second-generation bioethanol plants in a way that also facilitates recirculation of process water. PMID:19251882

Alriksson, Björn; Rose, Shaunita H.; van Zyl, Willem H.; Sjöde, Anders; Nilvebrant, Nils-Olof; Jönsson, Leif J.

2009-01-01

73

Production of glucose oxidase using Aspergillus niger and corn steep liquor.  

PubMed

Glucose oxidase production was optimized using an isolated strain of Aspergillus niger and an economical nutrient source, corn steep liquor (CSL). The culture produced 580 +/- 30 units/ml of the enzyme using 70 g/l sucrose as the carbon source. Using CSL as the sole nutrient source enzyme synthesis was increased to 640 +/- 36 units/ml. None of the nitrogen sources (nitrates of calcium, sodium, ammonium, potassium and yeast extract, malt extract, and peptone) was beneficial to the enzyme synthesis. Aeration and agitation enhanced enzyme synthesis to 850 +/- 45 units/ml. Glucose oxidase has numerous applications in food industry and clinical fields. PMID:11333029

Kona, R P; Qureshi, N; Pai, J S

2001-06-01

74

Biotransformation of the diperpenoid, isosteviol, by Aspergillus niger, Penicillium chrysogenum and Rhizopus arrhizus  

Microsoft Academic Search

The biotransformation of isosteviol (ent-16-ketobeyeran-19-oic acid) by three fungi is described. Aspergillus niger produced the 7?-OH derivative, ent-7?-hydroxy-16-ketobeyeran-19-oic, and the 1?,7?-diOH derivative, ent-1?,7?-dihydroxy-16-ketobeyeran-19-oic acid. The 17-OH compound, ent-17-hydroxy-16-ketobeyeran-19-oic acid, was obtained with Penicillium chrysogenum. Rhizopus arrhizus produced the 7?-OH derivative, ent-7?-hydroxy-16-ketobeyeran-19-oic acid. The isolated metabolites were characterised by IR, NMR and MS.

Brás H de Oliveira; Márcia C dos Santos; Paulo C Leal

1999-01-01

75

Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants  

Technology Transfer Automated Retrieval System (TEKTRAN)

Aspergillus niger and A. carbonarius are two species in the Aspergillus section Nigri (black-spored aspergilli) frequently associated with peanut (Arachis hypogea), maize (Zea mays), and other plants as pathogens. These infections are symptomless and as such are major concerns since some black aspe...

76

Biotransformation of the Streptomyces scabies phytotoxin thaxtomin A by the fungus Aspergillus niger.  

PubMed

Of several hundred microorganisms randomly selected from the environment, only a fungal isolate identified as Aspergillus niger van Tiegham var. niger was found to transform the phytotoxin thaxtomin A to much less toxic metabolites. The rate and extent of transformation of thaxtomin A was tested under a variety of conditions, including different growth media, biomass concentrations, incubation periods, and shaker speeds. Under optimum conditions the fungus converted thaxtomin A into two major and five minor metabolites. The two major metabolites and three of the five minor metabolites were fully characterized by a combination of mass spectral and nuclear magnetic resonance techniques. When assayed on aseptically produced mini-tubers, the major metabolites proved to be much less phytotoxic than thaxtomin A. PMID:15052314

Lazarovits, George; Hill, Jackie; King, Russell R; Calhoun, Larry A

2004-02-01

77

Induction of contour sensing in Aspergillus niger by stress and its relevance to fungal growth mechanics and hyphal tip structure  

E-print Network

Induction of contour sensing in Aspergillus niger by stress and its relevance to fungal growth to investigate the thigmotropic reactions of the ubiquitous fungus Aspergillus niger. This organism not appear to demonstrate thigmotropic growth under normal conditions. Our results show that A. niger undergoes significant

Davidson, Fordyce A.

78

Increased Heterologous Protein Production in Aspergillus niger Fermentation through Extracellular Proteases Inhibition by  

E-print Network

Increased Heterologous Protein Production in Aspergillus niger Fermentation through Extracellular in filamentous fungal fermentation and thereby to enhance heterologous protein production. Introduction with efficient heterologous protein production in the fungal fermentation industry (1, 2). Current strategies

Gu, Tingyue

79

Nanosulfur: A Potent Fungicide Against Food Pathogen, Aspergillus niger  

NASA Astrophysics Data System (ADS)

Elemental sulfur (S0), man's oldest eco-friendly fungicide for curing fungal infections in plants and animals, is registered in India as a non-systemic and contact fungicide. However due to its high volume requirement, Indian agrochemical industry and farmers could not effectively use this product till date. We hypothesize that intelligent nanoscience applications might increase the visibility of nanosulfur in Indian agriculture as a potent and eco-safe fungicide. Sulfur nanoparticles (NPs) were synthesized bottom-up via a liquid synthesis method with average particle size in the range of 50-80 nm and the shapes of the NPs were spherical. A comparative study of elemental and nano-sulfur produced has been tested against facultative fungal food pathogen, Aspergillus niger. Results showed that nanosulfur is more efficacious than its elemental form.

Choudhury, Samrat Roy; Nair, Kishore K.; Kumar, Rajesh; Gogoi, Robin; Srivastava, Chitra; Gopal, Madhuban; Subhramanyam, B. S.; devakumar, C.; Goswami, Arunava

2010-10-01

80

Nanosulfur: A Potent Fungicide Against Food Pathogen, Aspergillus niger  

SciTech Connect

Elemental sulfur (S{sup 0}), man's oldest eco-friendly fungicide for curing fungal infections in plants and animals, is registered in India as a non-systemic and contact fungicide. However due to its high volume requirement, Indian agrochemical industry and farmers could not effectively use this product till date. We hypothesize that intelligent nanoscience applications might increase the visibility of nanosulfur in Indian agriculture as a potent and eco-safe fungicide. Sulfur nanoparticles (NPs) were synthesized bottom-up via a liquid synthesis method with average particle size in the range of 50-80 nm and the shapes of the NPs were spherical. A comparative study of elemental and nano-sulfur produced has been tested against facultative fungal food pathogen, Aspergillus niger. Results showed that nanosulfur is more efficacious than its elemental form.

Choudhury, Samrat Roy; Goswami, Arunava [Agricultural and Ecological Research Unit, Biological Sciences Division, Indian Statistical Institute, 203 B. T. Road, Kolkata, West Bengal-700108 (India); Nair, Kishore K.; Kumar, Rajesh; Gopal, Madhuban; Devakumar, C. [Department of Agricultural Chemicals, Pusa Campus, New Delhi (India); Gogoi, Robin [Plant Pathology, Pusa Campus, New Delhi (India); Srivastava, Chitra; Subhramanyam, B. S. [Entomology, Indian Agricultural Research Institute, Pusa Campus, New Delhi (India)

2010-10-04

81

Biotransformation of germacranolide from Onopordon leptolepies by Aspergillus niger.  

PubMed

Terpenes are present in the essential oils obtained from herbs and spices. They are produced by these plant species as a chemical defense mechanism against phytopathogenic microorganisms. Therefore, terpenes have attracted great attention in the food industry, e.g., they have been used in foods such as cheese as natural preservatives to prevent fungal growth. Herein, we describe the microbial transformation of onopordopicrin (1) by Aspergillus niger. Four product 11? H-dihydroonopordopicrin (2), 11? H-dihydroonopordopicrin (3), 3?-hydroxy-11? H-dihydroonopordopicrin (4), and 14-hydroxy-11? H-dihydroonopordopicrin (5) were obtained. Their structures were identified on the basis of chemical and spectroscopic data. All the four compounds were novel. PMID:22186324

Esmaeili, Akbar; Moazami, Nasrin; Rustaiyan, Abdolhossein

2012-01-01

82

New pathway for the biodegradation of indole in Aspergillus niger  

SciTech Connect

Indole and its derivatives form a class of toxic recalcitrant environmental pollutants. The growth of Aspergillus niger was inhibited by very low concentrations (0.005 to 0.02%) of indole, even when 125- to 500-fold excess glucose was present in the medium. When 0.02% indole was added, the fungus showed a lag phase for about 30 h and the uptake of glucose was inhibited. Indole was metabolized by a new pathway via indoxyl (3-hydroxyindole), N-formylanthranilic acid, anthranilic acid, 2,3-dihydroxybenzoic acid, and catechol, which was further degraded by an ortho cleavage. The enzymes N-formylanthranilate deformylase, anthranilate hydroxylase, 2,3-dihydroxybenzoate decarboxylase, and catechol dioxygenase were induced by indole as early as after 5 h of growth, and their activities were demonstrated in a cell-free system.

Kamath, A.; Vaidyanathan, C.S. (Indiana Institute of Science, Bangalore (India))

1990-01-01

83

New pathway for the biodegradation of indole in Aspergillus niger.  

PubMed Central

Indole and its derivatives form a class of toxic recalcitrant environmental pollutants. The growth of Aspergillus niger was inhibited by very low concentrations (0.005 to 0.02%) of indole, even when 125- to 500-fold excess glucose was present in the medium. When 0.02% indole was added, the fungus showed a lag phase for about 30 h and the uptake of glucose was inhibited. Indole was metabolized by a new pathway via indoxyl (3-hydroxyindole), N-formylanthranilic acid, anthranilic acid, 2,3-dihydroxybenzoic acid, and catechol, which was further degraded by ortho cleavage. The enzymes N-formylanthranilate deformylase, anthranilate hydroxylase, 2,3-dihydroxybenzoate decarboxylase, and catechol dioxygenase were induced by indole as early as after 5 h of growth, and their activities were demonstrated in a cell-free system. PMID:2310183

Kamath, A V; Vaidyanathan, C S

1990-01-01

84

In-silico analysis of Aspergillus niger beta-glucosidases  

NASA Astrophysics Data System (ADS)

Genomic data mining was carried out and revealed a total of seventeen ?-glucosidases in filamentous fungi Aspergillus niger. Two of them belonged to glycoside hydrolase family 1 (GH1) while the rest belonged to genes in family 3 (GH3). These proteins were then named according to the nomenclature as proposed by the International Union of Biochemistry (IUB), starting from the lowest pI and glycoside hydrolase family. Their properties were predicted using various bionformatic tools showing the presence of domains for signal peptide and active sites. Interestingly, one particular domain, PA14 (protective antigen) was present in four of the enzymes, predicted to be involved in carbohydrate binding. A phylogenetic tree grouped the two glycoside hydrolase families with GH1 and GH3 related organisms. This study showed that the various domains present in these ?-glucosidases are postulated to be crucial for the survival of this fungus, as supported by other analysis.

Yeo S., L.; Shazilah, K.; Suhaila, S.; Abu Bakar F., D.; Murad A. M., A.

2014-09-01

85

Identification of Genes Associated with Morphology in Aspergillus Niger by Using Suppression Subtractive Hybridization  

SciTech Connect

The morphology of citric acid production strains of Aspergillus niger is sensitive to a variety of factors including the concentration of manganese (Mn2+). Upon increasing the Mn2+ concentration in A. niger (ATCC 11414) cultures to 14 ppb or higher, the morphology switches from pelleted to filamentous, accompanied by a rapid decline in citric acid production. Molecular mechanisms through which Mn2+ exerts effects on morphology and citric acid production in A. niger have not been well defined, but our use of suppression subtractive hybridization has identified 22 genes responsive to Mn2+. Fifteen genes were differentially expressed when A. niger was grown in media containing 1000 ppb Mn2+ (filamentous form) and seven genes in 10 ppb Mn2+ (pelleted form). Of the fifteen filamentous-associated genes, seven are novel and eight share 47-100% identity to genes from other organisms. Five of the pellet-associated genes are novel, and the other two genes encode a pepsin-type protease and polyubiquitin. All ten genes with deduced functions are either involved in amino acid metabolism/protein catabolism or cell regulatory processes. Northern-blot analysis showed that the transcripts of all 22 genes were rapidly enhanced or suppressed by Mn2+. Steady-state mRNA levels of six selected filamentous associated genes remained high during five days of culture in a filamentous state and low under pelleted growth conditions. The opposite behavior was observed for four selected pellet-associated genes. The full-length cDNA of the filamentous-associated clone, Brsa-25 was isolated. Antisense expression of Brsa-25 permitted pelleted growth and increased citrate production at higher concentrations of Mn2+ than could be tolerated by the parent strain. The results suggest the involvement of the newly isolated genes in regulation of A. niger morphology.

Dai, Ziyu; Mao, Xingxue; Magnuson, Jon K.; Lasure, Linda L.

2004-04-01

86

Identification of genes associated with morphology in Aspergillus niger by using suppression subtractive hybridization.  

PubMed

The morphology of citric acid production strains of Aspergillus niger is sensitive to a variety of factors, including the concentration of manganese (Mn(2+)). Upon increasing the Mn(2+) concentration in A. niger (ATCC 11414) cultures to 14 ppb or higher, the morphology switches from pelleted to filamentous, accompanied by a rapid decline in citric acid production. The molecular mechanisms through which Mn(2+) exerts effects on morphology and citric acid production in A. niger cultures have not been well defined, but our use of suppression subtractive hybridization has identified 22 genes responsive to Mn(2+). Fifteen genes were differentially expressed when A. niger was grown in media containing 1,000 ppb of Mn(2+) (filamentous form), and seven genes were expressed in 10 ppb of Mn(2+) (pelleted form). Of the 15 filament-associated genes, seven are novel and eight share 47 to 100% identity with genes from other organisms. Five of the pellet-associated genes are novel, and the other two genes encode a pepsin-type protease and polyubiquitin. All 10 genes with deduced functions are either involved in amino acid metabolism-protein catabolism or cell regulatory processes. Northern blot analysis showed that the transcripts of all 22 genes were rapidly enhanced or suppressed by Mn(2+). Steady-state mRNA levels of six selected filament-associated genes remained high during 5 days of culture in a filamentous state and remained low under pelleted growth conditions. The opposite behavior was observed for four selected pellet-associated genes. The full-length cDNA of the filament-associated clone, Brsa-25, was isolated. Antisense expression of Brsa-25 permitted pelleted growth and increased citrate production at concentrations of Mn(2+) that were higher than the parent strain could tolerate. These results suggest the involvement of the newly isolated genes in the regulation of A. niger morphology. PMID:15066846

Dai, Ziyu; Mao, Xingxue; Magnuson, Jon K; Lasure, Linda L

2004-04-01

87

Identification of Genes Associated with Morphology in Aspergillus niger by Using Suppression Subtractive Hybridization  

PubMed Central

The morphology of citric acid production strains of Aspergillus niger is sensitive to a variety of factors, including the concentration of manganese (Mn2+). Upon increasing the Mn2+ concentration in A. niger (ATCC 11414) cultures to 14 ppb or higher, the morphology switches from pelleted to filamentous, accompanied by a rapid decline in citric acid production. The molecular mechanisms through which Mn2+ exerts effects on morphology and citric acid production in A. niger cultures have not been well defined, but our use of suppression subtractive hybridization has identified 22 genes responsive to Mn2+. Fifteen genes were differentially expressed when A. niger was grown in media containing 1,000 ppb of Mn2+ (filamentous form), and seven genes were expressed in 10 ppb of Mn2+ (pelleted form). Of the 15 filament-associated genes, seven are novel and eight share 47 to 100% identity with genes from other organisms. Five of the pellet-associated genes are novel, and the other two genes encode a pepsin-type protease and polyubiquitin. All 10 genes with deduced functions are either involved in amino acid metabolism-protein catabolism or cell regulatory processes. Northern blot analysis showed that the transcripts of all 22 genes were rapidly enhanced or suppressed by Mn2+. Steady-state mRNA levels of six selected filament-associated genes remained high during 5 days of culture in a filamentous state and remained low under pelleted growth conditions. The opposite behavior was observed for four selected pellet-associated genes. The full-length cDNA of the filament-associated clone, Brsa-25, was isolated. Antisense expression of Brsa-25 permitted pelleted growth and increased citrate production at concentrations of Mn2+ that were higher than the parent strain could tolerate. These results suggest the involvement of the newly isolated genes in the regulation of A. niger morphology. PMID:15066846

Dai, Ziyu; Mao, Xingxue; Magnuson, Jon K.; Lasure, Linda L.

2004-01-01

88

New PCR method to differentiate species in the Aspergillus niger aggregate  

Microsoft Academic Search

The DNA that encodes the 5.8S gene of the ribosomal RNA and the two intergenic spacers ITS1 and ITS2 of the two proposed type strains of the Aspergillus niger aggregate (A. niger and Aspergillus tubingensis) have been sequenced. By comparison of sequences we have found that both species could be differentiated by RsaI digestion of the PCR products of the

F. Accensi; J. Cano; L. Figuera; M. L. Abarca; F. J. Cabañes

1999-01-01

89

Mapping the polysaccharide degradation potential of Aspergillus niger  

PubMed Central

Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger. PMID:22799883

2012-01-01

90

Metabolic control analysis of Aspergillus niger L-arabinose catabolism.  

PubMed

A mathematical model of the L-arabinose/D-xylose catabolic pathway of Aspergillus niger was constructed based on the kinetic properties of the enzymes. For this purpose L-arabinose reductase, L-arabitol dehydrogenase and D-xylose reductase were purified using dye-affinity chromatography, and their kinetic properties were characterized. For the other enzymes of the pathway the kinetic data were available from the literature. The metabolic model was used to analyze flux and metabolite concentration control of the L-arabinose catabolic pathway. The model demonstrated that flux control does not reside at the enzyme following the intermediate with the highest concentration, L-arabitol, but is distributed over the first three steps in the pathway, preceding and following L-arabitol. Flux control appeared to be strongly dependent on the intracellular L-arabinose concentration. At 5 mM intracellular L-arabinose, a level that resulted in realistic intermediate concentrations in the model, flux control coefficients for L-arabinose reductase, L-arabitol dehydrogenase and L-xylulose reductase were 0.68, 0.17 and 0.14, respectively. The analysis can be used as a guide to identify targets for metabolic engineering aiming at either flux or metabolite level optimization of the L-arabinose catabolic pathway of A. niger. Faster L-arabinose utilization may enhance utilization of readily available organic waste containing hemicelluloses to be converted into industrially interesting metabolites or valuable enzymes or proteins. PMID:16321042

de Groot, Marco J L; Prathumpai, Wai; Visser, Jaap; Ruijter, George J G

2005-01-01

91

[Lipid metabolism in Aspergillus niger under conditions of heat shock].  

PubMed

The processes of lipid synthesis and decomposition in Aspergillus niger under conditions of heat shock (HS) were studied in a pulse-chase experiment with 14C-labeled sodium acetate. HS (60 min) resulted in the synthesis of phospholipids and sphingolipids intensified compared tothe.control, as was evident from incorporation of the labeled substrate. The same pattern was observed for neutral lipids, especially for triacylglycerides, while incorporation of the label into sterols remained almost the same. Further cultivation for 3 h in the medium without the labeled substrate resulted in a significant decrease of the label content in the membrane lipids of both the control and the experiment, although under HS conditions this decrease was much more pronounced, especially for phosphatidylcholines and phosphatidylethanolamines. A threefold increase of the label content in phosphatidic acids was observed only under HS conditions. These results indicate more intense metabolism of the membrane lipids under heat shock and suggest the degradation of the major cell phospholipids as the factor responsible for the increased level of phosphatidic acids in A. niger mycelium. PMID:25509390

Tereshina, V M; Memorskaia, A S; Kotlova, E R

2013-01-01

92

Biochemical characterisation of extracellular phytase ( myo -inositol hexakisphosphate phosphohydrolase) from a hyper-producing strain of Aspergillus niger van Teighem  

Microsoft Academic Search

Aspergillus niger van Teighem, isolated in our laboratory from samples of rotten wood logs, produced extracellular phytase having a high specific activity of 22,592 units (mg protein)-1 . The enzyme was purified to near homogeneity using ion-exchange and gel-filtration chromatography. The molecular properties of the purified enzyme suggested the native phytase to be oligomeric, with a molecular weight of 353 kDa,

Purva Vats; U. C. Banerjee

2005-01-01

93

Apple pomace: A potential substrate for citric acid production by Aspergillus niger  

Microsoft Academic Search

Apple pomace was used as a fsubstrate for citric acid production by five strains of Aspergillus niger. A. niger NRRL 567 produced the greatest amount of citric acid from apple pomace in the presence of 4% methanol. The yield was 88% based on the amount of sugar consumed.

Y. D. Hang; E. E. Woodams

1984-01-01

94

Optimization of medium composition for the production of glucosyltransferase by Aspergillus niger with response surface methodology  

Microsoft Academic Search

Aspergillus niger CCRC 31494 produced an extracellular glucosyltransferase (EC 2.4.1.24) with a high transglucosylating activity. For production of glucosyltransferase by A. niger, yeast extract was the best nitrogen source after cultivation for 7 days. Addition of minerals to the medium showed no significant increase in the production of glucosyltransferase. A significant decrease in the production of glycosyltransferase was obtained when

Shiow-Ling Lee; Wen-Chang Chen

1997-01-01

95

Development of a homologous transformation system for Aspergillus niger based on the pyrG gene  

Microsoft Academic Search

The development of a homologous transformation system for Aspergillus niger is described. The system is based on the use of an orotidine-5'-phosphate decarboxylase deficient mutant (pyrG) and a vector, pAB4-1, which contains the functional A. niger pyrG gene as a selection marker. Transformation of the A. niger pyrG mutant with pAB4-1 resulted in the appearance of stable Pyr+ transformants at

Wim van Hartingsveldt; Ineke E. Mattern; Cora M. J. Zeijl; Peter H. Pouwels; Cees A. M. J. J. Hondel

1987-01-01

96

Chemical stabilization of glucoamylase from Aspergillus niger against thermal inactivation.  

PubMed

The applicability of crosslinking an enzyme to an oxidized polysaccharide by reductive alkylation to enhance thermostability has been investigated for glucoamylase from Aspergillus niger. Direct covalent coupling of the enzyme to periodate-oxidized dextran in the presence of NaBH(3)CN results in a conjugate which has thermal properties similar to those of the native enzyme. Our working hypothesis postulates that enhancement of thermostability will result from rigidification of the protein's conformation subsequent to the formation of multiple covalent bonds between the protein and the support. On the basis of the known characteristics of glucoamylase from Aspergillus niger, it would seem necessary to introduce additional amino groups in the polypeptide chain of the protein. The incorporation of new amino groups was performed in two phases. First, the glycosidic part of glucoamylase was oxidized by periodate and the resulting aldehyde groups were reductively aminated by a diaminoalkane and NaBH(3)CIM. Secondly, additional amino groups were introduced on carboxyl functions into the previously aminated glucoamylase by a diaminoalkane and a water-soluble carbodiimide in the presence of maltose to protect the active site. The final derivative was then coupled to periodate-oxidized dextran T-70 in the presence of NaBH(3)CN. Starting with native glucoamylase, three successive operations give rise to a conjugate which retained 27% of the initial activity when measured with soluble starch and 39% when measured with maltopentaose. Using substrates of various sizes, it was observed that steric hindrance at the active site may result from covalent coupling to dextran T-70. It was demonstrated in heat inactivation experiments that the thermostability of the conjugate was in all cases superior to that of the native enzymes. Finally, it was observed that the operational stability of the conjugate was at least twice that of native glucoamylase at 70 degrees C on 18% maltodextrin. Additional experiments rule out the possibility that thermosta-bilization of the complex is due to other reasons than the increase in the amino content of the protein prior to crosslinking. Neither chemical modification, reticulation nor change in the net charge of the protein resulted in a derivative of glucoamylase which presented enhanced thermostability after conjugation. We conclude that for enzymes which have a low content of available amino groups, the thermostabilization method proposed previously by the present authors may still be applicable if additional amino groups are introduced into the protein prior to its crosslinking to an oxidized polysaccharide. This new example reinforces the generality of this method of stabilization. PMID:18584602

Lenders, J P; Crichton, R R

1988-02-20

97

Facile production of Aspergillus niger ?-N-acetylgalactosaminidase in yeast.  

PubMed

?-N-Acetylgalactosaminidase (?-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal ?-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the ?-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae. The ?-GalNAc-ase gene contains an open reading frame which encodes a protein of 487 amino acid residues. The molecular mass of the mature protein deduced from the amino acid sequence of this reading frame is 54 kDa. The recombinant protein was purified to apparent homogeneity and biochemically characterized (pI4.4, K(M) 0.56 mmol/l for 2-nitrophenyl 2-acetamido-2-deoxy-?-d-galactopyranoside, and optimum enzyme activity was achieved at pH2.0-2.4 and 50-55°C). Its molecular weight was determined by analytical ultracentrifuge measurement and dynamic light scattering. Our experiments confirmed that the recombinant ?-GalNAc-ase exists as two distinct species (70 and 130 kDa) compared to its native form, which is purely monomeric. N-Glycosylation was confirmed at six of the eight potential N-glycosylation sites in both wild type and recombinant ?-GalNAc-ase. PMID:21982820

Mrázek, Hynek; Benada, Old?ich; Man, Petr; Van?k, Ond?ej; K?en, Vladimír; Bezouška, Karel; Weignerová, Lenka

2012-01-01

98

Some factors affecting tannase production by Aspergillus niger Van Tieghem  

PubMed Central

One variable at a time procedure was used to evaluate the effect of qualitative variables on the production of tannase from Aspergillus niger Van Tieghem. These variables including: fermentation technique, agitation condition, tannins source, adding carbohydrates incorporation with tannic acid, nitrogen source type and divalent cations. Submerged fermentation under intermittent shaking gave the highest total tannase activity. Maximum extracellular tannase activity (305 units/50 mL) was attained in medium containing tannic acid as tannins source and sodium nitrate as nitrogen source at 30 °C for 96 h. All added carbohydrates showed significant adverse effects on the production of tannase. All tested divalent cations significantly decreased tannase production. Moreover, split plot design was carried out to study the effect of fermentation temperature and fermentation time on tannase production. The results indicated maximum tannase production (312.7 units/50 mL) at 35 °C for 96 h. In other words, increasing fermentation temperature from 30 °C to 35 °C resulted in increasing tannase production. PMID:24294255

Aboubakr, Hamada A.; El-Sahn, Malak A.; El-Banna, Amr A.

2013-01-01

99

A two-step bioconversion process for vanillin production from ferulic acid combining Aspergillus niger and Pycnoporus cinnabarinus  

Microsoft Academic Search

A two-step bioconversion process of ferulic acid to vanillin was elaborated combining two filamentous fungi, Aspergillus niger and Pycnoporus cinnabarinus. In the first step, A. niger transformed ferulic acid to vanillic acid and in the second step vanillic acid was reduced to vanillin by P. cinnabarinus. Ferulic acid metabolism by A. niger occurred essentially via the propenoic chain degradation to

Laurence Lesage-Meessen; Michel Delattre; Mireille Haon; Jean-François Thibault; Benoit Colonna Ceccaldi; Pascal Brunerie; Marcel Asther

1996-01-01

100

Flavone Biotransformation by Aspergillus niger and the Characterization of Two Newly Formed Metabolites  

PubMed Central

Aspergillus niger isolated from Allium sativum was used at large scale fermentation (150 mg flavone/200 ml medium) to obtain suitable amounts of the products, efficient for identification. Then spectral analysis (UV, IR, 1H-NMR, 13C-NMR) and mass spectrometry were performed for the two products, which contributed to the identification process. The metabolite (1) was identified as 2'-hydroxydihydrochalcone, and the metabolite (2) was identified as 2'-hydroxyphenylmethylketone, which were more active than flavone itself. Antioxidant activities of the two isolated metabolites were tested compared with ascorbic acid. Antioxidant activity of metabolite (1) was recorded 64.58% which represented 79% of the antioxidant activity of ascorbic acid, and metabolite (2) was recorded 54.16% (67% of ascorbic acid activity). However, the antioxidant activity of flavone was recorded 37.50% which represented 46% of ascorbic acid activity. The transformed products of flavone have antimicrobial activity against Pseudomonas aeruginosa, Aspergillus flavus and Candida albicans, with MIC was recorded 250 µg/ml for metabolite (2) against all three organism and 500, 300, and 300 µg/ml for metabolite (1) against tested microorganisms (P. aeruginosa, Escherichia coli, Bacillus subtilis, and Klebsiella pneumonia, Fusarium moniliforme, A. flavus, Saccharomyces cerviceae, Kluveromyces lactis and C. albicans) at this order. PMID:23990746

Assawah, Suzan W.; El-Sharkawy, Saleh H.; Abdel-Salam, Amal

2008-01-01

101

Molecular detection of ochratoxigenic Aspergillus species isolated from coffee beans in Saudi Arabia.  

PubMed

Ten fungal isolates from coffee beans were morphologically identified as Aspergillus niger, A. ochraceus and A. carbonari-us (N = 5, 3, and 2, respectively). Only one isolate, morphologically identified as A. niger, was unable to produce ochratoxin A (OTA). This may be a new species in the Aspergillus section Nigri. OTA levels in all the other isolates were above the limit of detection (0.15 mg/kg). Based on microsatellite-primed PCR (MP-PCR) profiles, using three microsatellite primers, three main groups were obtained by UPGMA cluster analysis: A. niger, A. ochraceus and A. carbonarius. A clear-cut association was found between the MP-PCR genotype and the ability to produce OTA. Using the primer pairs OCRA1/OCRA2, a single fragment of about 400 bp was amplified only when genomic DNA from the A. ochraceus isolates was used. PMID:21128209

Moslem, M A; Mashraqi, A; Abd-Elsalam, K A; Bahkali, A H; Elnagaer, M A

2010-01-01

102

Improved transformation efficiency of Aspergillus niger using the homologous nia D gene for nitrate reductase  

Microsoft Academic Search

Aspergillus niger transformation frequencies of up to 1,176 transformants per µg DNA were achieved using the plasmid vector pSTA10 containing the A. niger nitrate reductase structural gene. Analysis of genomic endonuclease cleaved DNA from nitrate utilising transformants by DNA hybridisation, showed that most integration events are as a result of homologous recombination. The niaD transformation system was used successfully for

Edward I. Campbell; Shiela E. Unkles; Janet A. Macro; Cees van den Hondel; Roland Contreras; James R. Kinghorn

1989-01-01

103

Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88  

Microsoft Academic Search

The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level of synteny was observed with other aspergilli sequenced. Strong function predictions were made for 6,506 of

Herman J Pel; Johannes H de Winde; David B Archer; Paul S Dyer; Gerald Hofmann; Peter J Schaap; Geoffrey Turner; Ronald P de Vries; Richard Albang; Kaj Albermann; Mikael R Andersen; Jannick D Bendtsen; Jacques A E Benen; Marco van den Berg; Stefaan Breestraat; Mark X Caddick; Roland Contreras; Michael Cornell; Pedro M Coutinho; Etienne G J Danchin; Alfons J M Debets; Peter Dekker; Piet W M van Dijck; Alard van Dijk; Lubbert Dijkhuizen; Arnold J M Driessen; Christophe d'Enfert; Steven Geysens; Coenie Goosen; Gert S P Groot; Piet W J de Groot; Thomas Guillemette; Bernard Henrissat; Marga Herweijer; Johannes P T W van den Hombergh; Cees A M J J van den Hondel; Rene T J M van der Heijden; Rachel M van der Kaaij; Frans M Klis; Harrie J Kools; Christian P Kubicek; Patricia A van Kuyk; Jürgen Lauber; Xin Lu; Marc J E C van der Maarel; Rogier Meulenberg; Hildegard Menke; Martin A Mortimer; Jens Nielsen; Stephen G Oliver; Maurien Olsthoorn; Karoly Pal; Arthur F J Ram; Ursula Rinas; Johannes A Roubos; Cees M J Sagt; Monika Schmoll; Jibin Sun; David Ussery; Janos Varga; Wouter Vervecken; Holger Wedler; Han A B Wösten; An-Ping Zeng; Albert J J van Ooyen; Jaap Visser; Hein Stam

2007-01-01

104

Heterologous protein secretion from Aspergillus niger in phosphate-buffered batch culture  

Microsoft Academic Search

Aspergillus niger was grown in batch culture containing various initial concentrations of sodium phosphate buffer (pH 6.5). A wild-type strain of A. niger and a transformed strain producing hen egg-white lysozyme were studied. The maximum cell yield was attained in medium not supplemented with phosphate. In those cultures acidification of the medium resulted in a minimum of pH 2.0 before

David B. Archer; Ian N. Roberts; Donald A. MacKenzie

1990-01-01

105

Biosorption of phenol from an aqueous solution by Aspergillus niger biomass  

Microsoft Academic Search

Phenols in trace quantities are usually present in the treated effluent of many wastewater-treatment plants. Phenol contamination of drinking water even at 1 ?g\\/l concentration can cause significant taste and odor problems. This study investigates the use of non-viable pretreated cells of Aspergillus niger to remove phenol from an aqueous solution. Five types of non-viable pretreated A. niger biomass powders

J. R Rao; T Viraraghavan

2002-01-01

106

SORPTION OF HEAVY METALS BY THE SOIL FUNGI ASPERGILLUS NIGER AND MUCOR ROUXII  

EPA Science Inventory

Sorption of the nitrate salts of cadmium(II), copper (II), lanthanum(III) and silver (I) by two fungi, Aspergillus niger and Mucor rouxii, was evaluated using Fruendlich adsorption isotherms and energy dispersive X-ray electron microscopy. The linearized Freundlich isotherm descr...

107

Enzymatic Comparisons of Aspergillus niger PhyA and Escherichia coli AppA2 Phytases  

Technology Transfer Automated Retrieval System (TEKTRAN)

This study was to compare three phytase activity assays and kinetics of Aspergillus niger PhyA and Escherichia coli AppA2 phytases expressed in Pichia pastoris at the observed stomach pH of 3.5. In Experiment 1, equivalent phytase activities in the crude preparations of PhyA and AppA2 were tested ...

108

NIGERLYSINTM, HEMOLYSIN PRODUCED BY ASPERGILLUS NIGER, CAUSES LETHALITY OF PRIMARY RAT CORTICAL NEURONAL CELLS IN VITRO  

EPA Science Inventory

Aspergillus niger produced a proteinaceous hemolysin, nigerlysinTM when incubated on sheep's blood agar at both 23° C and 37°C. Nigerlysin was purified from tryptic soy broth culture filtrate. Purified nigerlysin has a molecular weight of approximately 72 kDa, with an...

109

The effect of the sugar source on citric acid production by Aspergillus niger  

Microsoft Academic Search

Under otherwise identical fermentation conditions, the sugar source has been shown to have a marked effect on citric acid production by Aspergillus niger. Sucrose was the most favourable source, followed by glucose and fructose and then lactose. No citric acid was produced from galactose. Strong relationships were observed between citric acid production and the activities of certain enzymes in myccelial

M. Hossain; J. D. Brooks; I. S. Maddox

1984-01-01

110

Systemic analysis of the response of Aspergillus niger to ambient pH  

Microsoft Academic Search

ABSTRACT: BACKGROUND: The filamentous fungus Aspergillus niger is an exceptionally efficient producer of organic acids, which is one of the reasons for its relevance to industrial processes and commercial importance. While it is known that the mechanisms regulating this production are tied to the levels of ambient pH, the reasons and mechanisms for this are poorly understood. METHODS: To cast

Mikael R Andersen; Linda Lehmann; Jens Nielsen

2009-01-01

111

Aspergillus niger DLFCC-90 Rhamnoside Hydrolase, a New Type of Flavonoid Glycoside Hydrolase  

PubMed Central

A novel rutin-?-l-rhamnosidase hydrolyzing ?-l-rhamnoside of rutin, naringin, and hesperidin was purified and characterized from Aspergillus niger DLFCC-90, and the gene encoding this enzyme, which is highly homologous to the ?-amylase gene, was cloned and expressed in Pichia pastoris GS115. The novel enzyme was classified in glycoside-hydrolase (GH) family 13. PMID:22544243

Liu, Tingqiang; Zhang, Chunzhi; Lu, Mingchun; Piao, Yongzhe; Ohba, Masashi; Tang, Minqian; Yuan, Xiaodong; Wei, Shenghua; Wang, Kan; Ma, Anzhou; Feng, Xue; Qin, Siqing; Mukai, Chisato; Tsuji, Akira

2012-01-01

112

Unfolding and Refolding of Aspergillus Niger PhyB Phytase: Role of Disulfide Bridges  

Technology Transfer Automated Retrieval System (TEKTRAN)

Role of disulfide bridges in folding of Aspergillus niger phytase pH 2.5-optimum (PhyB) was investigated using dynamic light scattering (DLS). Guanidinium chloride (GuCl) at 1.0 M unfolded phytase; however, its removal by dialysis refolded the protein. Thiol reagent, tris (2-carboxyethyl) phosphin...

113

Colonization of rye green manure and peanut fruit debris by Aspergillus falvus and Aspergillus niger group in field soils.  

PubMed Central

Aspergillus flavus and Aspergillus niger group colonization of deep-plowed, decomposing rye green manure cover crops in peanut field soils was studied in four fields during 1972 and 1973; colonization of decomposing peanut fruits was studied in 1972 in two fields. A. flavus colonization of rye and peanut fruits was greater in soils of heavy texture, and an A. flavus population as high as 165 propagules per g of soil was observed in soil adjacent to rye, whereas A. flavus populations in soils not associated with rye were 18 propagules per g of soil or lower. Highest A. flavus populations in soil adjacent to decomposing peanut fruits were usually comparable to populations associated with rye. Little decomposing rye or peanut fruit colonization was generally observed by the A. flavus competitor, A. niger group. A. flavus may maintain or increase its inoculum potential by colonization of these and other moribund plant tissues. PMID:823865

Griffin, G J; Garren, K H

1976-01-01

114

Correlation of mycotoxin fumonisin B2 production and presence of the fumonisin biosynthetic gene fum8 in Aspergillus niger from grape.  

PubMed

Aspergillus niger is a significant component of the fungal community on grapes. The mycotoxin fumonisin B2 (FB2) was recently detected in grape must and wine as well as in cultures of some A. niger strains isolated from grapes and raisins. This study examined 48 strains of Aspergillus section Nigri for the presence of the fumonisin biosynthetic gene fum8 in relation to FB2 production. The fum8 gene was detected in only 11 A. niger strains, 9 of which also produced FB2. Maximum parsimony analysis based on the calmodulin gene sequence indicated that the presence/absence of fum8 is not correlated with the phylogenetic relationship of the isolates. This is the first report correlating the presence of a fumonisin biosynthetic gene with fumonisin production in A. niger from an important food crop. The results suggest that the absence of FB2 production in grape isolates of A. niger can result from the absence of at least one gene essential for production. PMID:20666454

Susca, Antonia; Proctor, Robert H; Mulè, Giuseppina; Stea, Gaetano; Ritieni, Alberto; Logrieco, Antonio; Moretti, Antonio

2010-08-25

115

Phytase Production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through Submerged and Solid-State Fermentation  

PubMed Central

Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF) and solid-state fermentations (SSF) for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were capable of producing maximum phytase on 5th day of incubation in both submerged and solid-state fermentation media. Aspergillus niger CFR 335 and A. ficuum produced a maximum of 60.6?U/gds and 38?U/gds of the enzyme, respectively, in wheat bran solid substrate medium. Enhancement in the enzyme level (76 and 50.7?U/gds) was found when grown in a combined solid substrate medium comprising wheat bran, rice bran, and groundnut cake in the ratio of 2?:?1?:?1. A maximum of 9.6 and 8.2?U/mL of enzyme activity was observed in SmF by A. niger CFR 335 and A.ficuum, respectively, when grown in potato dextrose broth. PMID:24688383

Shivanna, Gunashree B.; Venkateswaran, Govindarajulu

2014-01-01

116

Overexpression of Aspergillus tubingensis faeA in protease-deficient Aspergillus niger enables ferulic acid production from plant material.  

PubMed

The production of ferulic acid esterase involved in the release of ferulic acid side groups from xylan was investigated in strains of Aspergillus tubingensis, Aspergillus carneus, Aspergillus niger and Rhizopus oryzae. The highest activity on triticale bran as sole carbon source was observed with the A. tubingensis T8.4 strain, which produced a type A ferulic acid esterase active against methyl p-coumarate, methyl ferulate and methyl sinapate. The activity of the A. tubingensis ferulic acid esterase (AtFAEA) was inhibited twofold by glucose and induced twofold in the presence of maize bran. An initial accumulation of endoglucanase was followed by the production of endoxylanase, suggesting a combined action with ferulic acid esterase on maize bran. A genomic copy of the A. tubingensis faeA gene was cloned and expressed in A. niger D15#26 under the control of the A. niger gpd promoter. The recombinant strain has reduced protease activity and does not acidify the media, therefore promoting high-level expression of recombinant enzymes. It produced 13.5 U/ml FAEA after 5 days on autoclaved maize bran as sole carbon source, which was threefold higher than for the A. tubingensis donor strain. The recombinant AtFAEA was able to extract 50 % of the available ferulic acid from non-pretreated maize bran, making this enzyme suitable for the biological production of ferulic acid from lignocellulosic plant material. PMID:24664515

Zwane, Eunice N; Rose, Shaunita H; van Zyl, Willem H; Rumbold, Karl; Viljoen-Bloom, Marinda

2014-06-01

117

New approach for selecting pectinase producing mutants of Aspergillus niger well adapted to solid state fermentation.  

PubMed

The aim of this paper is to review and study a new approach for improving strains of Aspergillus niger specially adapted to produce pectinases by Solid State Fermentation (SSF) with materials having low levels of water activity (a(w)), i.e., coffee pulp. Special emphasis is placed on the use of two antimetabolic compounds: 2-deoxy-glucose (DG) and 2,4-dinitro-phenol (DNP) combined with a water depressant (ethylene glycol = EG) in order to put strong selection pressures on UV treated spores from parental strain C28B25 isolated from a coffee plantation. Such a strain was found to be DG sensitive. Results suggested the existence of a reciprocal relation between adaptation of isolated strains to SSF or to Submerged Fermentation (SmF) systems. Preliminary physiological analysis of isolated strains showed that at least some few initially DG resistant mutants could revert to DG sensitive phenotype but conserving increased pectinase production. Also it was found that phenotype for DNP resistance could be associated to changes of DG resistance. Finally, it was found that low levels of a(w) produced by adding 15% EG to agar plates, were a significant selection factor for strains well adapted to SSF system. PMID:14545667

Antier, P; Minjares, A; Roussos, S; Viniegra-González, G

1993-01-01

118

Value addition of vegetable wastes by solid-state fermentation using Aspergillus niger for use in aquafeed industry.  

PubMed

Vegetable waste typically has high moisture content and high levels of protein, vitamins and minerals. Its value as an agricultural feed can be enhanced through solid-state fermentation (SSF). Two experiments were conducted to evaluate the nutritional status of the products derived by SSF of a mixture of dried vegetable waste powder and oil cake mixture (soybean flour, wheat flour, groundnut oil cake and sesame oil cake at 4:3:2:1 ratio) using fungi Aspergillus niger S(1)4, a mangrove isolate, and A. niger NCIM 616. Fermentation was carried out for 9 days at 35% moisture level and neutral pH. Significant (p<0.05) increase in crude protein and amino acids were obtained in both the trials. The crude fat and crude fibre content showed significant reduction at the end of fermentation. Nitrogen free extract (NFE) showed a gradual decrease during the fermentation process. The results of the study suggest that the fermented product obtained on days 6 and 9 in case of A. niger S(1)4 and A. niger NCIM 616 respectively contained the highest levels of crude protein. PMID:20100652

Rajesh, N; Imelda-Joseph; Raj, R Paul

2010-11-01

119

The effects of agitation and aeration on the production of gluconic acid by Aspergillus niger  

SciTech Connect

The effects of agitation and aeration in the production of gluconic acid by Aspergillus niger from a glucose medium were investigated. Experiments were conducted at aeration rates of 5.0 and 10.0 L/min. Four different agitation speeds were investigated for each aeration rate. Gluconic acid concentration and biomass concentration were analyzed, and the rate of consumption of substrate by A. niger was noted. The main purpose of this work was to find the optimal conditions of agitation and aeration for the growth of A. niger and production of gluconic acid in submerged culture in a batch fermentor at a bench-top scale. The oxygen-transfer rates at different agitation and aeration rates were calculated. The gluconic acid concentration and rate of growth of A. niger increased with increase in the agitation and aeration rates.

Dronawat, S.N.; Svihla, C.K.; Hanley, T.R. [Univ. of Louisville, KY (United States)

1995-12-31

120

Toolkit for visualization of the cellular structure and organelles in Aspergillus niger.  

PubMed

Aspergillus niger is a filamentous fungus that is extensively used in industrial fermentations for protein expression and the production of organic acids. Inherent biosynthetic capabilities, such as the capacity to secrete these biomolecules in high amounts, make A. niger an attractive production host. Although A. niger is renowned for this ability, the knowledge of the molecular components that underlie its production capacity, intercellular trafficking processes and secretion mechanisms is far from complete. Here, we introduce a standardized set of tools, consisting of an N-terminal GFP-actin fusion and codon optimized eforRed chromoprotein. Expression of the GFP-actin construct facilitates visualization of the actin filaments of the cytoskeleton, whereas expression of the chromoprotein construct results in a clearly distinguishable red phenotype. These experimentally validated constructs constitute the first set of standardized A. niger biomarkers, which can be used to study morphology, intercellular trafficking, and secretion phenomena. PMID:25524108

Buren, Emiel B J Ten; Karrenbelt, Michiel A P; Lingemann, Marit; Chordia, Shreyans; Deng, Ying; Hu, JingJing; Verest, Johanna M; Wu, Vincen; Gonzalez, Teresita J Bello; Heck, Ruben G A van; Odoni, Dorett I; Schonewille, Tom; Straat, Laura van der; Graaff, Leo H de; Passel, Mark W J van

2014-12-19

121

VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture.  

PubMed

Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ?veA strain. The latter implies a fourfold reduced surface area to develop chains of spores. Over and above this, the conidial chain length was approximately fivefold reduced. The calculated 20-fold reduction in formation of conidia by ?veA fits the 8- to 17-fold decrease in counted spore numbers. Notably, morphology of the ?veA conidiophores of A. niger was very similar to that of wild-type Aspergillus sydowii. This suggests that VeA is key in conidiophore architecture diversity in the fungal kingdom. The finding that biomass formation of the A. niger ?veA strain was reduced twofold shows that VeA not only impacts dispersion capacity but also colonization capacity of A. niger. PMID:25367340

Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

2015-01-01

122

The intra- and extracellular proteome of Aspergillus niger growing on defined medium with xylose or maltose as carbon substrate  

Microsoft Academic Search

BACKGROUND: The filamentous fungus Aspergillus niger is well-known as a producer of primary metabolites and extracellular proteins. For example, glucoamylase is the most efficiently secreted protein of Aspergillus niger, thus the homologous glucoamylase (glaA) promoter as well as the glaA signal sequence are widely used for heterologous protein production. Xylose is known to strongly repress glaA expression while maltose is

Xin Lu; Jibin Sun; Manfred Nimtz; Josef Wissing; An-Ping Zeng; Ursula Rinas

2010-01-01

123

Purification and characterization of a ferulic acid esterase (FAE-III) from Aspergillus niger: specificity for the phenolic moiety and binding to microcrystalline cellulose  

Microsoft Academic Search

An inducible ferulic acid esterase (FAE-Ill) has been isolated, purified and partially characterized from Aspergillus niger after growth on oat spelt xylan. The purification procedure utilized ammonium sulphate precipitation, hydrophobic interaction and anion-exchange chromatography. The purified enzyme appeared almost pure by SDS-PAGE, with an apparent M, of 36000. A single band, corresponding to a pl of 3.3 was observed on

Craig B. Faulds; Gary Williamson

1994-01-01

124

Enzymatic resolution of racemic phenyloxirane by a novel epoxide hydrolase from Aspergillus niger SQ6 and its fed-batch fermentation  

Microsoft Academic Search

A microorganism with the ability to catalyze the resolution of racemic phenyloxirane was isolated and identified as Aspergillus niger SQ-6. Chiral capillary electrophoresis was successfully applied to separate both phenyloxirane and phenylethanediol. The\\u000a epoxide hydrolase (EH) involved in this resolution process was (R)-stereospecific and constitutively expressed. When whole cells were used during the biotransformation process, the optimum\\u000a temperature and pH

Yanbin Liu; Qian Sha; Sheng Wu; Jianjun Wang; Liu Yang; Wanru Sun

2006-01-01

125

The promoter of the glucoamylase-encoding gene of Aspergillus niger functions in Ustilago maydis.  

PubMed

Promoter sequences from the Aspergillus niger glucoamylase-encoding gene (glaA) were linked to the bacterial hygromycin (Hy) phosphotransferase-encoding gene (hph) and this chimeric marker was used to select Hy-resistant (HyR) Ustilago maydis transformants. This is an example of an Ascomycete promoter functioning in a Basidiomycete. HyR transformants varied with respect to copy number of integrated vector, mitotic stability, and tolerance to Hy. Only 216 bp of glaA promoter sequence is required for expression in U. maydis but this promoter is not induced by starch as it is in Aspergillus spp. The transcriptional start points are the same in U. maydis and A. niger. PMID:2112106

Smith, T L; Gaskell, J; Berka, R M; Yang, M; Henner, D J; Cullen, D

1990-04-16

126

Comparing phosphorus mobilization strategies using Aspergillus niger for the mineral dissolution of three phosphate rocks.  

PubMed

Phosphorus deficiencies are limiting crop production in agricultural soils worldwide. Locally available sources of raw phosphate rock (PR) are being recognized for their potential role in soil fertility improvement. Phosphorus bioavailability is essential for the efficiency of PRs and can be increased by acid treatments. The utilization of organic acid producing micro-organisms, notably Aspergillus niger, presents a sustainable alternative to the use of strong inorganic acids, but acid production of A. niger strongly depends on the mineral content of the growth media. This study compared the phosphorus mobilization efficiency of two biological treatments, namely addition of acidic cell-free supernatants from A. niger cultivations to PRs and the direct cultivation of A. niger with PRs. The results show that addition of PR to cultivations leads to significant differences in the profile of organic acids produced by A. niger. Additions of PR, especially igneous rocks containing high amounts of iron and manganese, lead to reduced citric acid concentrations. In spite of these differences, phosphorus mobilization was similar between treatments, suggesting that the simpler direct cultivation method was not inferior. In addition to citric acid, it is suggested that oxalic acid contributes to PR solubilization in direct cultivations with A. niger, which would benefit farmers in developing countries where conventional fertilizers are not adequately accessible. PMID:19709342

Schneider, K D; van Straaten, P; de Orduña, R Mira; Glasauer, S; Trevors, J; Fallow, D; Smith, P S

2010-01-01

127

Cell Bound and Extracellular Glucose Oxidases from Aspergillus niger BTL: Evidence for a Secondary Glycosylation Mechanism  

Microsoft Academic Search

Two glucose oxidase (GOX) isoforms where purified to electrophoretic homogeneity from the mycelium extract (GOXI) and the extracellular medium (GOXII) of Aspergillus niger BTL cultures. Both enzymes were found to be homodimers with nonreduced molecular masses of 148 and 159 kDa and pI values\\u000a of 3.7 and 3.6 for GOXI and GOXII, respectively. The substrate specificity and the kinetic characteristics of

Dimitris G. Hatzinikolaou; Diomi Mamma; Paul Christakopoulos; Dimitris Kekos

2007-01-01

128

Screening, mutagenesis and protoplast fusion of Aspergillus niger for the enhancement of extracellular glucose oxidase production  

Microsoft Academic Search

Various strains of Aspergillus niger were screened for extracellular glucose oxidase (GOD) activity. The most effective producer, strain FS-3 (15.9 U mL?1), was mutagenized using UV-irradiation or ethyl methane sulfonate. Of the 400 mutants obtained, 32 were found to be resistant to 2-deoxy d-glucose, and 17 of these exhibited higher GOD activities (from 114.5 to 332.1%) than the original FS-3 strain. Following

A. A. Khattab; W. A. Bazaraa

2005-01-01

129

Lipase production from Aspergillus niger by solid-state fermentation using gingelly oil cake  

Microsoft Academic Search

Cultural conditions for the production of lipase by Aspergillus niger strain MTCC 2594 by solid-state fermentation using gingelly oil cake were standardized. A lipase activity of 363·6 U\\/g of dry substrate was obtained at 72 h under optimum conditions. Addition of various nitrogen sources, carbohydrates and inducers to the substrate was found to be ineffective. The enzyme was optimally active

N. R. Kamini; J. G. S. Mala; R. Puvanakrishnan

1998-01-01

130

Bioleaching of spent refinery processing catalyst using Aspergillus niger with high-yield oxalic acid  

Microsoft Academic Search

A spent refinery processing catalyst was physically and chemically characterized, and subjected to one-step and two-step bioleaching processes using Aspergillus niger. During bioleaching of the spent catalysts of various particle sizes (“as received”, 100–150?m, <37?m, and x¯=2.97 (average) ?m) and pulp densities, the biomass dry weight and pH were determined. The corresponding leach liquor was analysed for excreted organic acids

Deenan Santhiya; Yen-Peng Ting

2005-01-01

131

Removal of Congo Red from an aqueous solution by fungus Aspergillus niger  

Microsoft Academic Search

Biosorption is becoming a promising alternative to replace or supplement the present dye removal processes from dye wastewater. In this study, removal of an anionic disazo direct dye, Congo Red, from an aqueous solution by biosorption on dead fungus, Aspergillus niger, was investigated. Pretreatment with NaHCO3 was found to be the most effective with a biosorption capacity of 14.72 mg\\/g

Yuzhu Fu; T. Viraraghavan

2002-01-01

132

Expression of an Aspergillus niger Phytase Gene (phyA )i n Saccharomyces cerevisiae  

Microsoft Academic Search

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals and reduces phosphorus pollution of animal waste. Our objectives were to express an Aspergillus niger phytase gene (phyA )i nSaccharomyces cerevisiae and to determine the effects of glycosylation on the phytase's activity and thermostability. A 1.4-kb DNA fragment containing the coding region of the phyA gene

YANMING HAN; DAVID B. WILSON; XIN GEN LEI

1999-01-01

133

The role of the tricarboxylic acid cycle in citric acid accumulation by Aspergillus niger  

Microsoft Academic Search

Determinations of the momentary levels of various intermediates related to the activity of the tricarboxylic acid cycle have been made during citric acid production in high-accumulating (manganese deficient) and lowaccumulating (manganese supplemented) mycelia of Aspergillus niger. During the growth period the levels of almost all TCA cycle acids, with the exception of 2-oxo-acids, were unusually high; during the induction phase

C. P. Kubicek; M. Röhr

1978-01-01

134

Morphological development of Aspergillus niger immobilized in Ca-alginate and K-carrageenan  

Microsoft Academic Search

The morphological development of citric acid producing Aspergillus niger immobilized in Ca-alginate and K-carrageenan was studied. The fungus normally developed a dense mycelium layer below and on the gel bead surfaces so that substrate and oxygen in this area had direct contact with mycelia. By this way mycelia are not only immobilized by entrapment but also in a “pellet-like” matter.

H. Eikmeier; F. Westmeier; H. J. Rehm

1984-01-01

135

Nitrogen, carbon, and pH regulation of extracellular acidic proteases of Aspergillus niger  

Microsoft Academic Search

Aspergillus niger secretes a number of enzymes, including proteases, into its culture fluid. The regulation of the two major acidic extracellular proteases, pepA and pepB, was investigated using Northern analyses. Our data suggest that the regulation of pepA and pepB expression occurs predominantly at the level of mRNA content and that, while they are regulated in a similar manner, differences

Gabor Jarai; Frank Buxton

1994-01-01

136

The use of Aspergillus niger for the bioconversion of olive mill waste-waters  

Microsoft Academic Search

Olive mill waste-water was used for protein production in small-scale experiments, using non-sterilized medium without pH control. A 14 g\\/1 concentration of proteins, 61% chemical oxygen demand removal and a 58% reduction in total phenolic compounds were obtained using an Aspergillus niger strain. The removal of phenolic compounds resulted in a change in the colour of the waste-water from black

Moktar Hamdi; Abdelkader Khadir; Jean-Louis Garcia

1991-01-01

137

Sorption of heavy metals by the soil fungi 'Aspergillus niger' and Mucor rouxii  

Microsoft Academic Search

Sorption of the nitrate salts of cadmium(II), copper(II), lanthanum(III) and silver(I) by two fungi, Aspergillus niger and Mucor rouxii, was evaluated using Freundlich adsorption isotherms and energy dispersive X-ray electron microscopy. The linearized Freundlich isotherm described the metal sorption data well for metal concentrations of 5 microM-1 mM metal. Differences in metal binding were observed among metals, as well as

M. D. Mullen; D. C. Wolf; T. J. Beveridge; G. W. Bailey

1992-01-01

138

Production and characterization of a highly active cellobiase from Aspergillus niger grown in solid state fermentation  

Microsoft Academic Search

Summary  \\u000a Aspergillus niger produced extracellular cellobiase when grown on different lignocellulosic substrates in solid state fermentation. The enzyme activity and yield were variable according to the carbon source. In Vogel’s medium, the cellobiase productivity was significantly higher on wheat bran, followed by Leptochloa fusca (kallar grass) straw augmented with corn steep liquor. Maximum yield of cellobiase\\/g wheat bran was significantly

Muhammad Ibrahim Rajoka; Muhammad Waheed Akhtar; Atif Hanif; A. M. Khalid

2006-01-01

139

Citric acid production from carob pod extract by cell recycle of Aspergillus niger atcc 9142  

Microsoft Academic Search

The production of citric acid from carob pod extract by cell recycle of Aspergillus niger at different pHs was investigated. Best results in terms of citric acid concentration, productivity, yield and sugar utilization were obtained with a substrate pH of 5.0. The citric acid concentration (85.5 g\\/l) and the productivity (4 g\\/ld) remained constant up to the second and third

T. Roukas

1998-01-01

140

Sub-inhibitory concentration of biogenic selenium nanoparticles lacks post antifungal effect for Aspergillus niger and Candida albicans and stimulates the growth of Aspergillus niger  

PubMed Central

Background The antifungal activity of selenium nanoparticles (Se NPs) prepared by Klebsiella pneumoniae has been reported previously for different fungi. In the present study, freshly prepared Se NPs produced by K. pneumoniae were purified and characterized by transmission electron microscopy and Energy-Dispersive X-ray spectroscopy (EDS) and its post antifungal effects for two fungi were evaluated. Materials and Methods The minimum inhibitory concentrations (MICs) of Se NPs, determined by serial dilution were 250 µg/ml for Aspergillus niger and 2,000 µg/ml for Candida albicans. The effect of exposure of A. niger and C. albicans to Se NPs on later growth was evaluated by incubating the fungi for 1 hour at 25 °C in media containing 0, 1, 2 and 4 x MIC of Se NPs and diluting the cultures 100 times with Se free medium. The kinetics of growth of the fungi in control cultures and in non-toxic Se NPs concentration of, 0.01 × MIC, 0.02 × MIC or 0.04 × MIC were measured. Results The exposure of A. niger and C. albicans to 2 and 4 x MIC of Se NPs stimulated the growth of both fungi in the absence of toxic concentrations of Se. The strongest stimulation was observed for A. niger. Conclusion It is concluded that exposure to high concentration of the Se NPs did not have any post-inhibitory effect on A. niger and C. albicans and that trace amounts of this element promoted growth of both fungi in a dose- dependent-manner. The role of nanoparticles serving as needed trace elements and development of microorganism tolerance to nanoparticles should not be dismissed while considering therapeutic potential. PMID:23466957

Kazempour, Zahra Bahri; Yazdi, Mohammad Hossein; Rafii, Fatemeh; Shahverdi, Ahmad Reza

2013-01-01

141

Production of a bioflocculant from Aspergillus niger using palm oil mill effluent as carbon source.  

PubMed

This study evaluated the potential of bioflocculant production from Aspergillus niger using palm oil mill effluent (POME) as carbon source. The bioflocculant named PM-5 produced by A. niger showed a good flocculating capability and flocculating rate of 76.8% to kaolin suspension could be achieved at 60 h of culture time. Glutamic acid was the most favorable nitrogen source for A. niger in bioflocculant production at pH 6 and temperature 35 °C. The chemical composition of purified PM-5 was mainly carbohydrate and protein with 66.8% and 31.4%, respectively. Results showed the novel bioflocculant (PM-5) had high potential to treat river water from colloids and 63% of turbidity removal with the present of Ca(2+) ion. PMID:25189510

Aljuboori, Ahmad H Rajab; Uemura, Yoshimitsu; Osman, Noridah Binti; Yusup, Suzana

2014-11-01

142

Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88.  

PubMed

The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level of synteny was observed with other aspergilli sequenced. Strong function predictions were made for 6,506 of the 14,165 open reading frames identified. A detailed description of the components of the protein secretion pathway was made and striking differences in the hydrolytic enzyme spectra of aspergilli were observed. A reconstructed metabolic network comprising 1,069 unique reactions illustrates the versatile metabolism of A. niger. Noteworthy is the large number of major facilitator superfamily transporters and fungal zinc binuclear cluster transcription factors, and the presence of putative gene clusters for fumonisin and ochratoxin A synthesis. PMID:17259976

Pel, Herman J; de Winde, Johannes H; Archer, David B; Dyer, Paul S; Hofmann, Gerald; Schaap, Peter J; Turner, Geoffrey; de Vries, Ronald P; Albang, Richard; Albermann, Kaj; Andersen, Mikael R; Bendtsen, Jannick D; Benen, Jacques A E; van den Berg, Marco; Breestraat, Stefaan; Caddick, Mark X; Contreras, Roland; Cornell, Michael; Coutinho, Pedro M; Danchin, Etienne G J; Debets, Alfons J M; Dekker, Peter; van Dijck, Piet W M; van Dijk, Alard; Dijkhuizen, Lubbert; Driessen, Arnold J M; d'Enfert, Christophe; Geysens, Steven; Goosen, Coenie; Groot, Gert S P; de Groot, Piet W J; Guillemette, Thomas; Henrissat, Bernard; Herweijer, Marga; van den Hombergh, Johannes P T W; van den Hondel, Cees A M J J; van der Heijden, Rene T J M; van der Kaaij, Rachel M; Klis, Frans M; Kools, Harrie J; Kubicek, Christian P; van Kuyk, Patricia A; Lauber, Jürgen; Lu, Xin; van der Maarel, Marc J E C; Meulenberg, Rogier; Menke, Hildegard; Mortimer, Martin A; Nielsen, Jens; Oliver, Stephen G; Olsthoorn, Maurien; Pal, Karoly; van Peij, Noël N M E; Ram, Arthur F J; Rinas, Ursula; Roubos, Johannes A; Sagt, Cees M J; Schmoll, Monika; Sun, Jibin; Ussery, David; Varga, Janos; Vervecken, Wouter; van de Vondervoort, Peter J J; Wedler, Holger; Wösten, Han A B; Zeng, An-Ping; van Ooyen, Albert J J; Visser, Jaap; Stam, Hein

2007-02-01

143

Evidence for a cytoplasmic pathway of oxalate biosynthesis in Aspergillus niger  

SciTech Connect

Oxalate accumulation of up to 8 g/liter was induced in Aspergillus niger by shifting the pH from 6 to 8. This required the presence of P/sub i/ and a nitrogen source and was inhibited by the protein synthesis inhibitor cycloheximide. Exogenously added /sup 14/CO/sub 2/ was not incorporated into oxalate, but was incorporated into acetate and malate, thus indicating the biosynthesis of oxalate by hydrolytic cleavage of oxaloacetate. Inhibition of mitochondrial citrate metabolism by fluorocitrate did not significantly decrease the oxalate yield. The putative enzyme that was responsible for this oxaloacetate hydrolase (EC 3.7.1.1), which was induced de novo during the pH shift. Subcellular fractionation of oxalic acid-forming mycelia of A. niger showed that this enzyme is located in the cytoplasm of A. niger. The results are consistent with a cytoplasmic pathway of oxalate formation which does not involve the tricarboxylic acid cycle.

Kubicek, C.P.; Schreferl-Kunar, G.; Woehrer, W.; Roehr, M.

1988-03-01

144

Enantioselective behavior of lipases from Aspergillus niger immobilized in different supports.  

PubMed

Considering the extraordinary microbial diversity and importance of fungi as enzyme producers, the search for new biocatalysts with special characteristics and possible applications in biocatalysis is of great interest. Here, we report the performance in the resolution of racemic ibuprofen of a native enantioselective lipase from Aspergillus niger, free and immobilized in five types of support (Accurel EP-100, Amberlite MB-1, Celite, Montmorillonite K10 and Silica gel). Amberlite MB-1 was found to be the best support, with a conversion of 38.2%, enantiomeric excess of 50.7% and enantiomeric ratio (E value) of 19 in 72 h of reaction. After a thorough optimization of several parameters, the E value of the immobilized Aspergillus niger lipase was increased (E = 23) in a shorter reaction period (48 h) at 35 degrees C. Moreover, the immobilized Aspergillus niger lipase maintained an esterification activity of at least 80% after 8 months of storage at 4 degrees C and could be reused at least six times. PMID:19390883

da Silva, Vania Castriani Fernandes; Contesini, Fabiano Jares; de Oliveira Carvalho, Patrícia

2009-07-01

145

Biocatalytic potential of laccase-like multicopper oxidases from Aspergillus niger  

PubMed Central

Background Laccase-like multicopper oxidases have been reported in several Aspergillus species but they remain uncharacterized. The biocatalytic potential of the Aspergillus niger fungal pigment multicopper oxidases McoA and McoB and ascomycete laccase McoG was investigated. Results The laccase-like multicopper oxidases McoA, McoB and McoG from the commonly used cell factory Aspergillus niger were homologously expressed, purified and analyzed for their biocatalytic potential. All three recombinant enzymes were monomers with apparent molecular masses ranging from 80 to 110 kDa. McoA and McoG resulted to be blue, whereas McoB was yellow. The newly obtained oxidases displayed strongly different activities towards aromatic compounds and synthetic dyes. McoB exhibited high catalytic efficiency with N,N-dimethyl-p-phenylenediamine (DMPPDA) and 2,2-azino-di(3-ethylbenzthiazoline) sulfonic acid (ABTS), and appeared to be a promising biocatalyst. Besides oxidizing a variety of phenolic compounds, McoB catalyzed successfully the decolorization and detoxification of the widely used textile dye malachite green. Conclusions The A. niger McoA, McoB, and McoG enzymes showed clearly different catalytic properties. Yellow McoB showed broad substrate specificity, catalyzing the oxidation of several phenolic compounds commonly present in different industrial effluents. It also harbored high decolorization and detoxification activity with the synthetic dye malachite green, showing to have an interesting potential as a new industrial biocatalyst. PMID:23270588

2012-01-01

146

Induction of mutation in Aspergillus niger for conversion of cellulose into glucose  

SciTech Connect

Plant wastes are very important part of biomass used and investigated for energy, chemical, and fuel production. Cellulose is the major renewable form of carbohydrate in the world, about 10{sup 11} tons of which is synthesized annually. For general use, it must be hydrolyzed first, either chemically or by cellulases derived from a few specialized microorganisms. Enzymes are acceptable environmentally but expensive to produce. Certainly, induction of mutations and selection of high cellulose microbial strains with significant adaptability to degrade cellulose to glucose is promising solutions. Induction of mutations in other fungi and Aspergillus sp. rather than Aspergillus niger was reported. Aspergillus ustus and Trichoderma harzianum were induced by gamma irradiation indicating mutants that excrete higher cellulose yields, particularly exocellobiohydrolase (Avicelase) than their respective wild types. Mutants from the celluiolytic fungus Penicillium pinophilum were induced by chemical and UV-irradiation. Enhancing the production of endo-1,4-{Beta}-D-glucanase (CMCase) and particularly {Beta}-glucosidase was obtained by gamma irradiation of Altemaria alternate. To overcome the lower activity of {beta}-glucosidase in certain fungi species rather than A. niger, mixed cultures of different species were tried. Thus, Aspergillus phonicis with Trichoderma reesei Rut 30, produced a cellulose complex that improved activity twofold over cellulose from Trichoderma alone.

Helmi, S.; Khalil, A.E.; Tahoun, M.K.; Khairy, A.H. [Univ. of Alexandria Research Centre, Alexandria (Egypt)

1991-12-31

147

Spatial and Developmental Differentiation of Mannitol Dehydrogenase and Mannitol-1-Phosphate Dehydrogenase in Aspergillus niger?†  

PubMed Central

The presence of a mannitol cycle in fungi has been subject to discussion for many years. Recent studies have found no evidence for the presence of this cycle and its putative role in regenerating NADPH. However, all enzymes of the cycle could be measured in cultures of Aspergillus niger. In this study we have analyzed the localization of two enzymes from the pathway, mannitol dehydrogenase and mannitol-1-phosphate dehydrogenase, and the expression of their encoding genes in nonsporulating and sporulating cultures of A. niger. Northern analysis demonstrated that mpdA was expressed in both sporulating and nonsporulating mycelia, while expression of mtdA was expressed only in sporulating mycelium. More detailed studies using green fluorescent protein and dTomato fused to the promoters of mtdA and mpdA, respectively, demonstrated that expression of mpdA occurs in vegetative hyphae while mtdA expression occurs in conidiospores. Activity assays for MtdA and MpdA confirmed the expression data, indicating that streaming of these proteins is not likely to occur. These results confirm the absence of the putative mannitol cycle in A. niger as two of the enzymes of the cycle are not present in the same part of A. niger colonies. The results also demonstrate the existence of spore-specific genes and enzymes in A. niger. PMID:20305000

Aguilar-Osorio, Guillermo; vanKuyk, Patricia A.; Seiboth, Bernhard; Blom, Dirk; Solomon, Peter S.; Vinck, Arman; Kindt, Frits; Wösten, Han A. B.; de Vries, Ronald P.

2010-01-01

148

Biotransformation of ent-pimaradienoic acid by cell cultures of Aspergillus niger.  

PubMed

Microbial transformation stands out among the many possible semi-synthetic strategies employed to increase the variety of chemical structures that can be applied in the search for novel bioactive compounds. In this paper we obtained ent-pimaradienoic acid (1, PA, ent-pimara-8(14),15-dien-19-oic acid) derivatives by fungal biotransformation using Aspergillus niger strains. To assess the ability of such compounds to inhibit vascular smooth muscle contraction, we also investigated their spasmolytic effect, along with another five PA derivatives previously obtained in our laboratory, on aortic rings isolated from male Wistar rats. The microbial transformation experiments were conducted at 30°C using submerged shaken liquid culture (120 rpm) for 10 days. One known compound, 7?-hydroxy ent-pimara-8(14),15-dien-19-oic acid (2), and three new derivatives, 1?-hydroxy ent-pimara-6,8(14),15-trien-19-oic acid (3), 1?,6?,14?-trihydroxy ent-pimara-7,15-dien-19-oic acid (4), and 1?,6?,7?,11?-tetrahydroxy ent-pimara-8(14),15-dien-19-oic acid (5), were isolated and identified on the basis of spectroscopic analyses and computational studies. The compounds obtained through biotransformation (2-5) did not display a significant antispasmodic activity (values ranging from 0% to 16.8% of inhibition); however the previously obtained diterpene, methyl 7?-hydroxy ent-pimara-8(14),15-dien-19-oate (8), showed to be very effective (82.5% of inhibition). In addition, our biological results highlight the importance to study the antispasmodic potential of a large number of novel diterpenes, to conduct further structure-activity relationship investigations. PMID:23916147

Severiano, Marcela E; Simão, Marília R; Ramos, Henrique P; Parreira, Renato L T; Arakawa, Nilton S; Said, Suraia; Furtado, Niege A J C; de Oliveira, Dionéia C R; Gregório, Luis E; Tirapelli, Carlos R; Veneziani, Rodrigo C S; Ambrósio, Sérgio R

2013-09-15

149

Phylogenetic characterization and ochratoxin A--fumonisin profile of black Aspergillus isolated from grapes in Argentina.  

PubMed

Aspergillus section Nigri populations isolated from seven growing regions from Argentina were characterized by sequencing in order to identify species responsible for production of ochratoxin A (OTA) and fumonisins (FB(s)). Sequences of genes encoding calmodulin, ?-tubulin, the second largest subunit of RNA polymerase II and translation elongation factor 1 alpha were analysed. The phylogenetic analysis showed the presence of six lineages: A. carbonarius, A. tubingensis, A. niger, A. japonicus, A. homomorphus and A. foetidus grouped in four major clusters. The molecular tools used allowed the identification for the first time of A. homomorphus from vineyards. OTA production confirmed the importance of A. carbonarius as the main ochratoxigenic species isolated and, to a variable degree, of A. niger and A. tubingensis, which were by far the most commonly occurring species on grapes in Argentina. The only strains able to produce OTA and fumonisins (B(2)-B(4)) belong to the A. niger cluster. PMID:21723640

Chiotta, M L; Susca, A; Stea, G; Mulè, G; Perrone, G; Logrieco, A; Chulze, S N

2011-09-15

150

Mutualistic interaction between Salmonella enterica and Aspergillus niger and its effects on Zea mays colonization  

PubMed Central

Salmonella?Typhimurium inhabits a variety of environments and is able to infect a broad range of hosts. Throughout its life cycle, some hosts can act as intermediates in the path to the infection of others. Aspergillus niger is a ubiquitous fungus that can often be found in soil or associated to plants and microbial consortia. Recently, S. Typhimurium was shown to establish biofilms on the hyphae of A. niger. In this work, we have found that this interaction is stable for weeks without a noticeable negative effect on either organism. Indeed, bacterial growth is promoted upon the establishment of the interaction. Moreover, bacterial biofilms protect the fungus from external insults such as the effects of the anti-fungal agent cycloheximide. Thus, the Salmonella–Aspergillus interaction can be defined as mutualistic. A tripartite gnotobiotic system involving the bacterium, the fungus and a plant revealed that co-colonization has a greater negative effect on plant growth than colonization by either organism in dividually. Strikingly, co-colonization also causes a reduction in plant invasion by S. Typhimurium. This work demonstrates that S. Typhimurium and A. niger establish a mutualistic interaction that alters bacterial colonization of plants and affects plant physiology. PMID:25351041

Balbontín, Roberto; Vlamakis, Hera; Kolter, Roberto

2014-01-01

151

Aspergillus niger is a filamentous fungus that is ubiquitous and commonly found on decaying plant material. A. niger has a saprophytic lifestyle and plays an important role in  

E-print Network

Summary Aspergillus niger is a filamentous fungus that is ubiquitous and commonly into smaller molecules that can be taken up and serve as energy and nutrient sources, the fungus is therefore of great importance for future optimisation of heterologous protein production in the fungus

Hille, Sander

152

Gene cloning and soluble expression of Aspergillus niger phytase in E. coli cytosol via chaperone co-expression.  

PubMed

A phytase gene from Aspergillus niger was isolated and two Escherichia coli expression systems, based on T7 RNA polymerase promoter and tac promoter, were used for its recombinant expression. Co-expression of molecular chaperone, GroES/EL, aided functional cytosolic expression of the phytase in E. coli BL21 (DE3). Untagged and maltose-binding protein-tagged recombinant phytase showed an activity band of ~49 and 92 kDa, respectively, on a zymogram. Heterologously-expressed phytase was fractionated from endogenous E. coli phytase by (NH4)2SO4 precipitation. The enzyme had optimum activity at 50 °C and pH 6.5. PMID:24078121

Ushasree, Mrudula Vasudevan; Vidya, Jalaja; Pandey, Ashok

2014-01-01

153

Formation of Sclerotia and Production of Indoloterpenes by Aspergillus niger and Other Species in Section Nigri  

PubMed Central

Several species in Aspergillus section Nigri have been reported to produce sclerotia on well-known growth media, such as Czapek yeast autolysate (CYA) agar, with sclerotia considered to be an important prerequisite for sexual development. However Aspergillus niger sensu stricto has not been reported to produce sclerotia, and is thought to be a purely asexual organism. Here we report, for the first time, the production of sclerotia by certain strains of Aspergillus niger when grown on CYA agar with raisins, or on other fruits or on rice. Up to 11 apolar indoloterpenes of the aflavinine type were detected by liquid chromatography and diode array and mass spectrometric detection where sclerotia were formed, including 10,23-dihydro-24,25-dehydroaflavinine. Sclerotium induction can thus be a way of inducing the production of new secondary metabolites from previously silent gene clusters. Cultivation of other species of the black aspergilli showed that raisins induced sclerotium formation by A. brasiliensis, A. floridensis A. ibericus, A. luchuensis, A. neoniger, A. trinidadensis and A. saccharolyticus for the first time. PMID:24736731

Frisvad, Jens C.; Petersen, Lene M.; Lyhne, E. Kirstine; Larsen, Thomas O.

2014-01-01

154

Removal of silver nanoparticles using live and heat shock Aspergillus niger cultures.  

PubMed

Silver nanoparticles (SNPs) are extensively used in many industrial and medical applications; however, the impact of their release in the environment is still considered an understudied field. In the present work, SNPs present in aqueous lab waste water (average size of 30 nm) were used to determine their impact on microflora if released in soil rhizosphere and sewage waste water. The results showed that 24 h incubation with different SNP concentrations resulted in a 2.6-fold decrease for soil rhizosphere microflora and 7.45-fold decrease for sewage waste water microflora, both at 24 ppm. Live and heat shock (50 and 70 °C) Aspergillus niger cultures were used to remove SNP waste, the results show 76.6, 81.74 and 90.8 % SNP removal, respectively after 3 h incubation. There was an increase in the log total bacterial count again after SNP removal by A. niger in the following order: live A. niger < 50 °C heat shock A. niger < 70 °C heat shock A. niger. The pH value decreased from 5.8 to 3.8 in the same order suggesting the production of an acid in the culture media. Scanning electron microscopy images showed agglomeration and/or complexation of SNP particles, in a micron size, in between the fungal mycelia, hence settling on and in between the mycelial network. The results suggest that silver was reduced again and agglomerated and/or chelated together in its oxidized form by an acid in A. niger media. More studies are recommended to determine the acid and the heat shock proteins to confirm the exact mode of action. PMID:24415500

Gomaa, Ola M

2014-06-01

155

The transcriptomic fingerprint of glucoamylase over-expression in Aspergillus niger  

PubMed Central

Background Filamentous fungi such as Aspergillus niger are well known for their exceptionally high capacity for secretion of proteins, organic acids, and secondary metabolites and they are therefore used in biotechnology as versatile microbial production platforms. However, system-wide insights into their metabolic and secretory capacities are sparse and rational strain improvement approaches are therefore limited. In order to gain a genome-wide view on the transcriptional regulation of the protein secretory pathway of A. niger, we investigated the transcriptome of A. niger when it was forced to overexpression the glaA gene (encoding glucoamylase, GlaA) and secrete GlaA to high level. Results An A. niger wild-type strain and a GlaA over-expressing strain, containing multiple copies of the glaA gene, were cultivated under maltose-limited chemostat conditions (specific growth rate 0.1 h-1). Elevated glaA mRNA and extracellular GlaA levels in the over-expressing strain were accompanied by elevated transcript levels from 772 genes and lowered transcript levels from 815 genes when compared to the wild-type strain. Using GO term enrichment analysis, four higher-order categories were identified in the up-regulated gene set: i) endoplasmic reticulum (ER) membrane translocation, ii) protein glycosylation, iii) vesicle transport, and iv) ion homeostasis. Among these, about 130 genes had predicted functions for the passage of proteins through the ER and those genes included target genes of the HacA transcription factor that mediates the unfolded protein response (UPR), e.g. bipA, clxA, prpA, tigA and pdiA. In order to identify those genes that are important for high-level secretion of proteins by A. niger, we compared the transcriptome of the GlaA overexpression strain of A. niger with six other relevant transcriptomes of A. niger. Overall, 40 genes were found to have either elevated (from 36 genes) or lowered (from 4 genes) transcript levels under all conditions that were examined, thus defining the core set of genes important for ensuring high protein traffic through the secretory pathway. Conclusion We have defined the A. niger genes that respond to elevated secretion of GlaA and, furthermore, we have defined a core set of genes that appear to be involved more generally in the intensified traffic of proteins through the secretory pathway of A. niger. The consistent up-regulation of a gene encoding the acetyl-coenzyme A transporter suggests a possible role for transient acetylation to ensure correct folding of secreted proteins. PMID:23237452

2012-01-01

156

An Antifungal Role of Hydrogen Sulfide on the Postharvest Pathogens Aspergillus niger and Penicillium italicum  

PubMed Central

In this research, the antifungal role of hydrogen sulfide (H2S) on the postharvest pathogens Aspergillus niger and Penicillium italicum growing on fruits and under culture conditions on defined media was investigated. Our results show that H2S, released by sodium hydrosulfide (NaHS) effectively reduced the postharvest decay of fruits induced by A. niger and P. italicum. Furthermore, H2S inhibited spore germination, germ tube elongation, mycelial growth, and produced abnormal mycelial contractions when the fungi were grown on defined media in Petri plates. Further studies showed that H2S could cause an increase in intracellular reactive oxygen species (ROS) in A. niger. In accordance with this observation we show that enzyme activities and the expression of superoxide dismutase (SOD) and catalase (CAT) genes in A. niger treated with H2S were lower than those in control. Moreover, H2S also significantly inhibited the growth of Saccharomyces cerevisiae, Rhizopus oryzae, the human pathogen Candida albicans, and several food-borne bacteria. We also found that short time exposure of H2S showed a microbicidal role rather than just inhibiting the growth of microbes. Taken together, this study suggests the potential value of H2S in reducing postharvest loss and food spoilage caused by microbe propagation. PMID:25101960

Li, Yan-Hong; Hu, Liang-Bin; Yan, Hong; Liu, Yong-Sheng; Zhang, Hua

2014-01-01

157

Kinetics of cellobiose hydrolysis using cellobiase composites from Trichoderma reesei and Aspergillus niger  

SciTech Connect

The enzymatic hydrolysis of cellulose to glucose involves the formation of cellobiose as an intermediate. It has been found necessary to add cellobiase from Aspergillus niger (NOVO) to the cellobiase component of Trichoderma reesei mutant Rut C-30 (Natick) cellulase enzymes in order to obtain after 48 h complete conversion of the cellobiose formed in the enzymatic hydrolysis of biomass. This study of the cellobiase activity of these two enzyme sources was undertaken as a first step in the formation of a kinetic model for cellulose hydrolysis that can be used in process design. In order to cover the full range of cellobiose concentrations, it was necessary to develop separate kinetic parameters for high- and low-concentration ranges of cellobiose for the enzymes from each organism. Competitive glucose inhibition was observed with the enzymes from both organisms. Substrate inhibition was observed only with the A. niger enzymes.

Grous, W.; Converse, A.; Grethlein, H.; Lynd, L.

1985-01-01

158

Inhibition of Aspergillus niger Phosphate Solubilization by Fluoride Released from Rock Phosphate  

PubMed Central

The simultaneous release of various chemical elements with inhibitory potential for phosphate solubilization from rock phosphate (RP) was studied in this work. Al, B, Ba, Ca, F, Fe, Mn, Mo, Na, Ni, Pb, Rb, Si, Sr, V, Zn, and Zr were released concomitantly with P during the solubilization of Araxá RP (Brazil), but only F showed inhibitory effects on the process at the concentrations detected in the growth medium. Besides P solubilization, fluoride decreased fungal growth, citric acid production, and medium acidification by Aspergillus niger. At the maximum concentration found during Araxá RP solubilization (22.9 mg F? per liter), fluoride decreased P solubilization by 55%. These findings show that fluoride negatively affects RP solubilization by A. niger through its inhibitory action on the fungal metabolism. Given that fluoride is a common component of RPs, the data presented here suggest that most of the microbial RP solubilization systems studied so far were probably operated under suboptimal conditions. PMID:23770895

Mendes, Gilberto de Oliveira; Vassilev, Nikolay Bojkov; Bonduki, Victor Hugo Araújo; da Silva, Ivo Ribeiro; Ribeiro, José Ivo

2013-01-01

159

A comparative investigation on the bioaccumulation of heavy metal ions by growing Rhizopus arrhizus and Aspergillus niger  

Microsoft Academic Search

The bioaccumulation of Cu(II) and Cd(II) ions by viable Rhizopus arrhizus and Aspergillus niger was studied as a function of initial pH and initial metal ion concentration. Optimum pH values for maximum Cu(II) and Cd(II) accumulation were determined, respectively, as 4.5 and 3.5 for R. arrhizus and 5.0 and 3.0 for A. niger. Although both the metal ions caused an

A. Y. Dursun; G. Uslu; O. Tepe

2003-01-01

160

The transcription factor HACA mediates the unfolded protein response in Aspergillus niger , and up-regulates its own transcription  

Microsoft Academic Search

The unfolded protein response (UPR) involves a complex signalling pathway in which the transcription factor HACA plays a central role. Here we report the cloning and characterisation of the hacA gene and its product from Aspergillus niger. ER (endoplasmic reticulum) stress results in the splicing of an unconventional 20-nt intron from the A. niger hacA mRNA, and is associated with

H. J. Mulder; M. Saloheimo; M. Penttilä; S. M. Madrid

2004-01-01

161

The complete amino acid sequence of the glycoprotein, glucoamylase G1, from Aspergillus niger  

Microsoft Academic Search

The primary structure of glucoamylase G1 (EC 3.2.1.3) from Aspergillus niger has been determined. Fragments of G1 were obtained\\u000a by cleavage with either cyanogen bromide, hydroxylamine, or S. aureus V8 protease. The resulting peptides were separated using\\u000a ion exchange chromatography on DEAE-Sephacel, gel filtration, and affinity chromatography on Con A-Sepharose. Secondary fragments\\u000a were generated by cleavage with either o-iodosobenzoic acid

Birte Svensson; Kjeld Larsen; Ib Svendsen; Esper Boel

1983-01-01

162

Purification and biochemical characterization of a novel thermostable lipase from Aspergillus niger  

Microsoft Academic Search

An extracellular 1,3-specific lipase with molecular weight of 35.5 kDa and an isoelectric point of 4.4 from Aspergillus niger has been purified 50-fold by pH precipitation followed by a series of chromatographic steps with an overall yield of 10%.\\u000a The enzyme was homogeneous as judged by denaturing polyacrylamide gel electrophoresis and size-exclusion fast-performance\\u000a liquid chromatography. It contained 2.8% sugar which

Variketta M. Haridasan Namboodiri; Rajagopal Chattopadhyaya

2000-01-01

163

Gene overexpression and biochemical characterization of the biotechnologically relevant chlorogenic acid hydrolase from Aspergillus niger.  

PubMed

The full-length gene that encodes the chlorogenic acid hydrolase from Aspergillus niger CIRM BRFM 131 was cloned by PCR based on the genome of the strain A. niger CBS 513.88. The complete gene consists of 1,715 bp and codes for a deduced protein of 512 amino acids with a molecular mass of 55,264 Da and an acidic pI of 4.6. The gene was successfully cloned and overexpressed in A. niger to yield 1.25 g liter(-1), i.e., 330-fold higher than the production of wild-type strain A. niger CIRM BRFM131. The histidine-tagged recombinant ChlE protein was purified to homogeneity via a single chromatography step, and its main biochemical properties were characterized. The molecular size of the protein checked by mass spectroscopy was 74,553 Da, suggesting the presence of glycosylation. ChlE is assembled in a tetrameric form with several acidic isoforms with pIs of around 4.55 and 5.2. Other characteristics, such as optimal pH and temperature, were found to be similar to those determined for the previously characterized chlorogenic acid hydrolase of A. niger CIRM BRFM 131. However, there was a significant temperature stability difference in favor of the recombinant protein. ChlE exhibits a catalytic efficiency of 12.5 x 10(6) M(-1) s(-1) toward chlorogenic acid (CGA), and its ability to release caffeic acid from CGA present in agricultural by-products such as apple marc and coffee pulp was clearly demonstrated, confirming the high potential of this enzyme. PMID:17630312

Benoit, Isabelle; Asther, Michèle; Bourne, Yves; Navarro, David; Canaan, Stéphane; Lesage-Meessen, Laurence; Herweijer, Marga; Coutinho, Pedro M; Asther, Marcel; Record, Eric

2007-09-01

164

Comparative genomics of citric-acid producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88  

SciTech Connect

The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compels additional exploration. We therefore undertook whole genome sequencing of the acidogenic A. niger wild type strain (ATCC 1015), and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was utilized to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 megabase of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis revealed up-regulation of the electron transport chain, specifically the alternative oxidative pathway in ATCC 1015, while CBS 513.88 showed significant up-regulation of genes relevant to glucoamylase A production, such as tRNA-synthases and protein transporters. Our results and datasets from this integrative systems biology analysis resulted in a snapshot of fungal evolution and will support further optimization of cell factories based on filamentous fungi.[Supplemental materials (10 figures, three text documents and 16 tables) have been made available. The whole genome sequence for A. niger ATCC 1015 is available from NBCI under acc. no ACJE00000000. The up-dated sequence for A. niger CBS 513.88 is available from EMBL under acc. no AM269948-AM270415. The sequence data from the phylogeny study has been submitted to NCBI (GU296686-296739). Microarray data from this study is submitted to GEO as series GSE10983. Accession for reviewers is possible through: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi token GSE10983] The dsmM_ANIGERa_coll511030F library and platform information is deposited at GEO under number GPL6758

Grigoriev, Igor V.; Baker, Scott E.; Andersen, Mikael R.; Salazar, Margarita P.; Schaap, Peter J.; Vondervoot, Peter J.I. van de; Culley, David; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristen F.; Albang, Richard; Albermann, Kaj; Berka, Randy M.; Braus, Gerhard H.; Braus-Stromeyer, Susanna A.; Corrochano, Luis M.; Dai, Ziyu; Dijck, Piet W.M. van; Hofmann, Gerald; Lasure, Linda L.; Magnusson, Jon K.; Meijer, Susan L.; Nielsen, Jakob B.; Nielsen, Michael L.; Ooyen, Albert J.J. van; Panther, Kathyrn S.; Pel, Herman J.; Poulsen, Lars; Samson, Rob A.; Stam, Hen; Tsang, Adrian; Brink, Johannes M. van den; Atkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Kubicek, Christian P.; Martinez, Diego; Peij, Noel N.M.E. van; Roubos, Johannes A.; Nielsen, Jens

2011-04-28

165

Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88  

PubMed Central

The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compel additional exploration. We therefore undertook whole-genome sequencing of the acidogenic A. niger wild-type strain (ATCC 1015) and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence, and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was used to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 Mb of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis supported up-regulation of genes associated with biosynthesis of amino acids that are abundant in glucoamylase A, tRNA-synthases, and protein transporters in the protein producing CBS 513.88 strain. Our results and data sets from this integrative systems biology analysis resulted in a snapshot of fungal evolution and will support further optimization of cell factories based on filamentous fungi. PMID:21543515

Andersen, Mikael R.; Salazar, Margarita P.; Schaap, Peter J.; van de Vondervoort, Peter J.I.; Culley, David; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristian F.; Albang, Richard; Albermann, Kaj; Berka, Randy M.; Braus, Gerhard H.; Braus-Stromeyer, Susanna A.; Corrochano, Luis M.; Dai, Ziyu; van Dijck, Piet W.M.; Hofmann, Gerald; Lasure, Linda L.; Magnuson, Jon K.; Menke, Hildegard; Meijer, Martin; Meijer, Susan L.; Nielsen, Jakob B.; Nielsen, Michael L.; van Ooyen, Albert J.J.; Pel, Herman J.; Poulsen, Lars; Samson, Rob A.; Stam, Hein; Tsang, Adrian; van den Brink, Johannes M.; Atkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Grigoriev, Igor V.; Kubicek, Christian P.; Martinez, Diego; van Peij, Noël N.M.E.; Roubos, Johannes A.; Nielsen, Jens; Baker, Scott E.

2011-01-01

166

Comparative genomics of citric-acid producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88  

SciTech Connect

The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compels additional exploration. We therefore undertook whole genome sequencing of the acidogenic A. niger wild type strain (ATCC 1015), and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was utilized to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 megabase of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis revealed up-regulation of the electron transport chain, specifically the alternative oxidative pathway in ATCC 1015, while CBS 513.88 showed significant up regulation of genes associated with biosynthesis of amino acids that are abundant in glucoamylase A, tRNA-synthases and protein transporters.

Andersen, Mikael R.; Salazar, Margarita; Schaap, Peter; van de Vondervoort, Peter; Culley, David E.; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristian F.; Albang, Richard; Albermann, Kaj; Berka, Randy; Braus, Gerhard; Braus-Stromeyer, Susanna A.; Corrochano, Luis; Dai, Ziyu; van Dijck, Piet; Hofmann, Gerald; Lasure, Linda L.; Magnuson, Jon K.; Menke, Hildegard; Meijer, Martin; Meijer, Susan; Nielsen, Jakob B.; Nielsen, Michael L.; van Ooyen, Albert; Pel, Herman J.; Poulsen, Lars; Samson, Rob; Stam, Hein; Tsang, Adrian; van den Brink, Johannes M.; ATkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Grigoriev, Igor V.; Kubicek, Christian P.; Martinez, Diego; van Peij, Noel; Roubos, Johannes A.; Nielsen, Jens B.; Baker, Scott E.

2011-06-01

167

Effect of oxygen transfer rate on the composition of the pectolytic enzyme complex of Aspergillus niger  

SciTech Connect

Optimal agitation and aeration conditions (assuring O/sub 2/ transfer rates (OTR) of 12-179 mmol/L-h) were determined for pectin lyase (PL) synthesis of an Aspergillus niger strain. Components of the pectolytic enzyme complex were also investigated in order to determine whether their O/sub 2/ demand is identical with or different from that of pectin lyase. Should the latter be the case, a possibility would be given to produce enzyme complexes of different agitation and aeration conditions. The mycelium yield of Aspergillus niger was maximum at an OTR of 100 mmol/L-h. The yields of the various pectolytic enzymes reached maximum at different OTRs. PL production was highest (0.555 mumol/min-mL) at an OTR of 60 mmol/L-h. Endopolygalacturonase (PG) production has a maximum at OTR 49 mmol/L-h, with a 2nd peak at 100-135 mmol O2/L-h. Pectin esterase (PE) synthesis showed a maximum at an OTR of 12-14 mmol/L-h, while both apple juice clarifying and macerating activities gave 2 maximum at 14 and 60 mmol/L-h due to the optima of PE and endo-PG. Macerating activity showed a high value at OTR optimal for PL production as well.

Zetelaki-Horvath, K.; Vas, K.

1981-01-01

168

Isolation and identification of Aspergillus spp. from brown kiwi (Apteryx mantelli) nocturnal houses in New Zealand.  

PubMed

Aspergillosis, a disease caused by infection with Aspergillus spp., is a common cause of death in birds globally and is an irregular cause of mortality of captive kiwi (Apteryx spp.). Aspergillus spp. are often present in rotting plant material, including the litter and nesting material used for kiwi in captivity. The aim of this study was to survey nocturnal kiwi houses in New Zealand to assess the levels of Aspergillus currently present in leaf litter. Samples were received from 11 nocturnal kiwi houses from throughout New Zealand, with one site supplying multiple samples over time. Aspergillus was isolated and quantified by colony counts from litter samples using selective media and incubation temperatures. Isolates were identified to the species level by amplification and sequencing of ITS regions of the ribosomal. Aspergillus spp. were recovered from almost every sample; however, the levels in most kiwi houses were below 1000 colony-forming units (CFU)/g of wet material. The predominant species was Aspergillus fumigatus, with rare occurrences of Aspergillus niger, Aspergillus nidulans, and Aspergillus parasiticus. Only one site had no detectable Aspergillus. The limit of detection was around 50 CFU/g wet material. One site was repeatedly sampled as it had a high loading of A. fumigatus at the start of the survey and had two recent clinical cases of aspergillosis diagnosed in resident kiwi. Environmental loading at this site with Aspergillus spp. reduced but was not eliminated despite changes of the litter. The key finding of our study is that the background levels of Aspergillus spores in kiwi nocturnal houses in New Zealand are low, but occasional exceptions occur and are associated with the onset of aspergillosis in otherwise healthy birds. The predominant Aspergillus species present in the leaf litter was A. fumigatus, but other species were also present. Further research is needed to confirm the optimal management of leaf litter to minimize Aspergillus spore counts. However, in the interim, our recommendations are that leaf litter should be freshly collected from areas of undisturbed forest areas and spread immediately after collection, without interim storage. PMID:24758108

Glare, Travis R; Gartrell, Brett D; Brookes, Jenny J; Perrott, John K

2014-03-01

169

Efficacy and possible mechanisms of perillaldehyde in control of Aspergillus niger causing grape decay.  

PubMed

A variety of plant products have been recognized for their antifungal activity and recently have attracted food industry attention for their efficacy in controlling postharvest fungal decay of fruits. The antifungal activity of perillaldehyde (PAE) was evaluated against Aspergillus niger, a known cause of grape spoilage, and possible mechanisms were explored. PAE showed notable antifungal activity against A. niger, with a minimum inhibitory concentration (MIC) and a minimum fungicidal concentration (MFC) of 0.25 and 1?l/ml, respectively. The accumulation of mycelial biomass was also inhibited by PAE in a dose-dependent manner, completely inhibiting mycelial growth at 1?l/ml. In vivo data confirmed that the vapour treatment of grapes with various concentrations of PAE markedly improved control of A. niger and suppressed natural decay. Concentrations of PAE of 0.075?l/ml air showed the greatest inhibition of fungal growth compared to the controls. Further experiments indicated that PAE activated a membrane-active mechanism that inhibits ergosterol synthesis, increases membrane permeability (as evidenced by extracellular pH and conductivity measurements), and disrupts membrane integrity, leading to cell death. Our findings suggest that this membrane-active mechanism makes PAE a promising potential antifungal agent for postharvest control of grape spoilage. PMID:25755082

Tian, Jun; Wang, Yanzhen; Zeng, Hong; Li, Zongyun; Zhang, Peng; Tessema, Akalate; Peng, Xue

2015-06-01

170

EglC, a New Endoglucanase from Aspergillus niger with Major Activity towards Xyloglucan  

PubMed Central

A novel gene, eglC, encoding an endoglucanase, was cloned from Aspergillus niger. Transcription of eglC is regulated by XlnR, a transcriptional activator that controls the degradation of polysaccharides in plant cell walls. EglC is an 858-amino-acid protein and contains a conserved C-terminal cellulose-binding domain. EglC can be classified in glycoside hydrolase family 74. No homology to any of the endoglucanases from Trichoderma reesei was found. In the plant cell wall xyloglucan is closely linked to cellulose fibrils. We hypothesize that the EglC cellulose-binding domain anchors the enzyme to the cellulose chains while it is cleaving the xyloglucan backbone. By this action it may contribute to the degradation of the plant cell wall structure together with other enzymes, including hemicellulases and cellulases. EglC is most active towards xyloglucan and therefore is functionally different from the other two endoglucanases from A. niger, EglA and EglB, which exhibit the greatest activity towards ?-glucan. Although the mode of action of EglC is not known, this enzyme represents a new enzyme function involved in plant cell wall polysaccharide degradation by A. niger. PMID:11916668

Hasper, Alinda A.; Dekkers, Ester; van Mil, Marc; van de Vondervoort, Peter J. I.; de Graaff, Leo H.

2002-01-01

171

Deletion of a Chitin Synthase Gene in a Citric Acid Producing Strain of Aspergillus niger  

SciTech Connect

Citric acid production by the filamentous fungus Aspergillus niger is carried out in a process that causes the organism to drastically alter its morphology. This altered morphology includes hyphal swelling and highly limited polar growth resulting in clumps of swollen cells that eventually aggregate into pellets of approximately 100 microns in diameter. In this pelleted form, A. niger has increased citric acid production as compared to growth in filamentous form. Chitin is a crucial component of the cell wall of filamentous fungi. Alterations in the deposition or production of chitin may have profound effects on the morphology of the organism. In order to study the role of chitin synthesis in pellet formation we have deleted a chitin synthase gene (csmA) in Aspergillus niger strain ATCC 11414 using a PCR based deletion construct. This class of chitin synthases is only found in filamentous fungi and is not present in yeasts. The csmA genes contain a myosin motor domain at the N-terminus and a chitin synthesis domain at the C-terminus. They are believed to contribute to the specialized polar growth observed in filamentous fungi that is lacking in yeasts. The csmA deletion strain (csmA?) was subjected to minimal media with and without osmotic stabilizers as well as tested in citric acid production media. Without osmotic stabilizers, the mutant germlings were abnormally swollen, primarily in the subapical regions, and contained large vacuoles. However, this swelling is ultimately not inhibitory to growth as the germlings are able to recover and undergo polar growth. Colony formation was largely unaffected in the absence of osmotic stabilizers. In citric acid production media csmA? was observed to have a 2.5 fold increase in citric acid production. The controlled expression of this class of chitin synthases may be useful for improving production of organic acids in filamentous fungi.

Rinker, Torri E.; Baker, Scott E.

2007-01-29

172

Radiosensitization of Aspergillus niger and Penicillium chrysogenum using basil essential oil and ionizing radiation for food decontamination.  

Technology Transfer Automated Retrieval System (TEKTRAN)

The Minimum Inhibitory Concentration (MIC) of basil oil, was determined for two pathogenic fungi of rice, Aspergillus niger and Penicillium chrysogenum. The antifungal activity of the basil oil in combination with ionising radiation was then investigated to determine if basil oil caused radiosensit...

173

Comparison of different inoculating methods to evaluate the pathogenicity and virulence of Aspergillus niger on two maize hybrids  

Technology Transfer Automated Retrieval System (TEKTRAN)

A two-year field study was conducted to determine the effects of inoculation techniques on the aggressiveness of Aspergillus niger kernel infection in A. flavus resistant and susceptible maize hybrids. Ears were inoculated with the silk-channel, side-needle, and spray techniques 7 days after midsilk...

174

Regulation of citric acid production by oxygen: Effect of dissolved oxygen tension on adenylate levels and respiration in Aspergillus niger  

Microsoft Academic Search

The mechanism of the control of citric acid accumulation by oxygen was investigated by means of pilot plant fermentation using Aspergillus niger. The critical dissolved oxygen tension (DOT) for oxygen uptake of this fungus was about 18–21 and 23–26 mbar for trophophase and idiophase, respectively. Minimal DOT for citric acid production was about 25 mbar. Citric acid production increased steadily

C. P. Kubicek; O. Zehentgruber; Housam El-Kalak; M. Röhr

1980-01-01

175

Presence of epoxide hydrolase activity in Aspergillus niger: Hydrolysis of 6', 7'-epoxybergamottin to 6', 7'-dihydroxybergamottin  

Technology Transfer Automated Retrieval System (TEKTRAN)

The 6', 7'-epoxybergamottin (EB) is one of major furanocoumarins in grapefruit. Previously, we have shown that Aspergillus niger has a capability of metabolizing EB into 6', 7'-dihydroxybergamottin (DHB), which is further metabolized to bergaptol and bergaptol-5-sulfate in vivo. In this study, we at...

176

Properties of epoxide hydrolase from Aspergillus niger for the hydrolytic kinetic resolution of epoxides in pure organic media  

Microsoft Academic Search

The effects of various reaction parameters regarding activity, stability and apparent enantioselectivity of recombinant epoxide hydrolase (EH) from Aspergillus niger in organic media were investigated using the racemic para-chlorostyrene oxide as a model substrate. Depending on the nature of the organic solvent, EH responded differently to variations in initial aw and no correlation between the activity or enantiomeric ratio (E)

S. Karboune; A. Archelas; J. Baratti

2006-01-01

177

Decolorization and detoxification of Synozol red HF-6BN azo dye, by Aspergillus niger and Nigrospora sp  

PubMed Central

In the present investigation the fungi, Aspergillus niger and Nigrospora sp. were employed for decolorization of Synozol red HF-6BN. Decolorization study showed that Aspergillus niger and Nigrospora sp. were able to decolorize 88% and 96% Synozol red 6BN, respectively, in 24 days. It was also studied that 86% and 90% Synozol red containing of dye effluent was decolorized by Aspergillus niger and Nigrospora sp. after 28 days of incubation at room temperature. A fungal-based protein with relative molecular mass of 70 kDa was partially purified and examined for enzymatic characteristics. The enzyme exhibited highest activity at temperature ranging from 40-50°C and at pH=6.0. The enzyme activity was enhanced in the presence of metal cations. High performance liquid chromatography analysis confirmed that these fungal strains are capable to degrade Synozol red dye into metabolites. No zones of inhibition on agar plates and growth of Vigna radiata in the presence of dye extracted sample, indicated that the fungal degraded dye metabolites are nontoxic to beneficial micro-flora and plant growth. Aspergillus niger and Nigrospora sp. have promising potential in color removal from textile wastewater-containing azo dyes. PMID:23369298

2013-01-01

178

Morphology engineering - Osmolality and its effect on Aspergillus niger morphology and productivity  

PubMed Central

Background The filamentous fungus Aspergillus niger is a widely used strain in a broad range of industrial processes from food to pharmaceutical industry. One of the most intriguing and often uncontrollable characteristics of this filamentous organism is its complex morphology, ranging from dense spherical pellets to viscous mycelia depending on culture conditions. Optimal productivity correlates strongly with a specific morphological form, thus making high demands on process control. Results In about 50 2L stirred tank cultivations the influence of osmolality on A. niger morphology and productivity was investigated. The specific productivity of fructofuranosidase producing strain A. niger SKAn 1015 could be increased notably from 0.5 to 9 U mg-1 h-1 around eighteen fold, by increasing the culture broth osmolality by addition of sodium chloride. The specific productivity of glucoamylase producing strain A. niger AB1.13, could be elevated using the same procedure. An optimal producing osmolality was shown to exist well over the standard osmolality at about 3.2 osmol kg-1 depending on the strain. Fungal morphology of all cultivations was examined by microscope and characterized by digital image analysis. Particle shape parameters were combined to a dimensionless Morphology number, which enabled a comprehensive characterization of fungal morphology correlating closely with productivity. A novel method for determination of germination time in submerged cultivations by laser diffraction, introduced in this study, revealed a decelerated germination process with increasing osmolality. Conclusions Through the introduction of the versatile Morphology number, this study provides the means for a desirable characterization of fungal morphology and demonstrates its relation to productivity. Furthermore, osmolality as a fairly new parameter in process engineering is introduced and found to affect fungal morphology and productivity. Osmolality might provide an auspicious and reliable approach to increase the productivity in industrial processes. Because of the predictable behavior fungal morphology showed in dependence of osmolality, a customization of morphology for process needs seems feasible. PMID:21801352

2011-01-01

179

Cytosolic streaming in vegetative mycelium and aerial structures of Aspergillus niger  

PubMed Central

Aspergillus niger forms aerial hyphae and conidiophores after a period of vegetative growth. The hyphae within the mycelium of A. niger are divided by septa. The central pore in these septa allows for cytoplasmic streaming. Here, we studied inter- and intra-compartmental streaming of the reporter protein GFP in A. niger. Expression of the gene encoding nuclear targeted GFP from the gpdA or glaA promoter resulted in strong fluorescence of nuclei within the vegetative hyphae and weak fluorescence in nuclei within the aerial structures. These data and nuclear run on experiments showed that gpdA and glaA are higher expressed in the vegetative mycelium when compared to aerial hyphae, conidiophores and conidia. Notably, gpdA or glaA driven expression of the gene encoding cytosolic GFP resulted in strongly fluorescent vegetative hyphae and aerial structures. Apparently, GFP streams from vegetative hyphae into aerial structures. This was confirmed by monitoring fluorescence of photo-activatable GFP (PA-GFP). In contrast, PA-GFP did not stream from aerial structures to vegetative hyphae. Streaming of PA-GFP within vegetative hyphae or within aerial structures of A. niger occurred at a rate of 10–15 ?m s-1. Taken together, these results not only show that GFP streams from the vegetative mycelium to aerial structures but it also indicates that its encoding RNA is not streaming. Absence of RNA streaming would explain why distinct RNA profiles were found in aerial structures and the vegetative mycelium by nuclear run on analysis and micro-array analysis. PMID:23450745

Bleichrodt, R.; Vinck, A.; Krijgsheld, P.; van Leeuwen, M.R.; Dijksterhuis, J.; Wösten, H.A.B.

2013-01-01

180

Three new species of Aspergillus section Flavi isolated from almonds and maize in Portugal  

Technology Transfer Automated Retrieval System (TEKTRAN)

Three new aflatoxin-producing species belonging to Aspergillus section Flavi are described, Aspergillus mottae, Aspergillus sergii and Aspergillus transmontanensis. These species were isolated from Portuguese almonds and maize. An investigation examining morphology, extrolites and molecular data was...

181

Treatment of poplar alkaline peroxide mechanical pulping (APMP) effluent with Aspergillus niger.  

PubMed

Although the moderate load (COD of 5000-10,000 mg/L) and biodegradability of the APMP pulping effluent should allow biological treatment, toxic compounds in the effluent can interfere with this type of treatment. Studies were conducted to determine if treatment of the effluent with Aspergillus niger S13 was feasible. Under the optimized conditions (3% inoculum, pH 6, shaking at 160 rpm, 60-72 h, and 30°C), this fungus was able to remove about 97% of the methyl tertiary butyl ether (MTBE) extractives, and 60%, 77% and 43% of the chemical oxygen demand, turbidity and color even without a pre-flocculation step. These results are of practical interest in China because the APMP process has become popular, and efficient and cost-effective effluent treatment technologies are in high demand. PMID:21612920

Liu, Tingzhi; Hu, Huiren; He, Zhibin; Ni, Yonghao

2011-08-01

182

Refinement of the crystal structures of biomimetic weddellites produced by microscopic fungus Aspergillus niger  

NASA Astrophysics Data System (ADS)

The single-crystal structures of four biomimetic weddellites CaC2O4 · (2 + x)H2O with different contents of zeolitic water ( x = 0.10-0.24 formula units) produced by the microscopic fungus Aspergillus niger were refined from X-ray diffraction data ( R = 0.029-0.038). The effect of zeolitic water content on the structural stability of weddellite was analyzed. The parameter a was shown to increase with increasing x due to the increase in the distance between water molecules along this direction. The water content and structural parameters of the synthesized weddellites are similar to those of weddellites from biofilms and kidney stones.

Rusakov, A. V.; Frank-Kamenetskaya, O. V.; Gurzhiy, V. V.; Zelenskaya, M. S.; Izatulina, A. R.; Sazanova, K. V.

2014-05-01

183

An acidic pectin lyase from Aspergillus niger with favourable efficiency in fruit juice clarification.  

PubMed

The pectin lyase gene pnl-zj5a from Aspergillus niger ZJ5 was identified and expressed in Pichia pastoris. PNL-ZJ5A was purified by ultrafiltration, anion exchange and gel chromatography. The Km and Vmax values determined using citrus pectin were 0.66 mg ml(-1) and 32.6 ?mol min(-1) mg(-1) , respectively. PNL-ZJ5A exhibited optimal activity at 43°C and retained activity over 25-50°C. PNL-ZJ5A was optimally active at pH 5 and effective in apple juice clarification. Compared with controls, PNL-ZJ5A increased the fruit juice yield significantly. Furthermore, PNL-ZJ5A reduced the viscosity of apple juice by 38.8% and increased its transmittance by 86.3%. PNL-ZJ5A combined with a commercial pectin esterase resulted in higher juice volume. PMID:25382689

Xu, S X; Qin, X; Liu, B; Zhang, D Q; Zhang, W; Wu, K; Zhang, Y H

2015-02-01

184

Optimization of enantioselective resolution of racemic ibuprofen by native lipase from Aspergillus niger.  

PubMed

Resolution of (R,S)-ibuprofen (2-(4-isobutylphenyl)propionic acid) enantiomers by esterification reaction with 1-propanol in different organic solvents was studied using native Aspergillus niger lipase. The main variables controlling the process (enzyme concentration and 1-propanol:ibuprofen molar ratio) have been optimized using response surface methodology based on a five-level, two-variable central composite rotatable design, in which the selected objective function was enantioselectivity. This enzyme preparation showed preferentially catalyzes the esterification of R(-)-ibuprofen, and under optimum conditions (7% w/v of enzyme and molar ratio of 2.41:1) the enantiomeric excess of active S(+)-ibuprofen and total conversion values were 79.1 and 48.0%, respectively, and the E-value was 32, after 168 h of reaction in isooctane. PMID:16680456

de O Carvalho, Patrícia; Contesini, Fabiano J; Bizaco, Renato; Calafatti, Silvana Ap; Macedo, Gabriela A

2006-08-01

185

Selection of amylolytically active Aspergillus niger mutants to 2-deoxy-D-glucose.  

PubMed

As a result of mutagenization and passaging on 2-deoxy-D-glucose containing medium, 10 Aspergillus niger strains resistant to this agent were obtained. These showed (with one exception) an increase in the activity of glucoamylse, the level of which ranged widely in individual cases from several to over 200% in comparison with the parent strain. A weaker rate of glucose accumulation in derepressed strains may account for the fact that the mechanism of their resistance to deoxyglucose is connected with disturbance of the system of glucose transport. However, it is possible that a high activity of acid phosphatase, which the obtained deoxyglucose-resistant cultures showed, may be involved here. Apart from the biochemical character of the catabolic derepression, it seems that it can already be successfully utilized to increase the productivity of industrial mould cultures. PMID:3122460

Fiedurek, J; Paszczy?ski, A; Ginalska, G; Ilczuk, Z

1987-01-01

186

Naphtho-?-pyrones from Endophyte Aspergillus niger occurring in the liverwort Heteroscyphus tener (Steph.) Schiffn.  

PubMed

Bioactivity-guided fractionation of the cytotoxic extract of Aspergillus niger, an endophytic fungus from the Chinese liverwort Heteroscyphus tener (Steph.) Schiffn., afforded five new naphtho-?-pyrones, rubrofusarin-6-O-?-D-ribofuranoside (1), (R)-10-(3-succinimidyl)-TMC-256A1 (2), asperpyrone E (3), isoaurasperone A (4), and isoaurasperone F (5), as well as four known ones, dianhydroaurasperone C (6), aurasperone D (7), asperpyrone D (8), and asperpyrone A (9), together with a cytotoxic cyclic pentapeptide, malformin A1 (10). Their structures were determined by extensive spectroscopic analysis. The absolute configurations of dimeric naphtho-?-pyrones 3-9 were also determined by analysis of their respective CD spectra. PMID:23847065

Li, Xiao-Bin; Xie, Fei; Liu, Shan-Shan; Li, Ying; Zhou, Jin-Chuan; Liu, Yong-Qing; Yuan, Hui-Qing; Lou, Hong-Xiang

2013-07-01

187

Influence of culture conditions on the biotransformation of (+)-limonene by Aspergillus niger.  

PubMed

The influence of the cultivation system and of the culture medium on the biotransformation of (+)-limonene by a strain of Aspergillus niger was investigated. Biooxidation products were obtained in all conditions tested. Using a laboratory bioreactor, six terpenes were identified in every medium, predominantly terpineols and carveols, whereas terpinen-4-ol and perillyl alcohol were the only terpenes found when flasks were used for culture. Perillyl alcohol and carveols predominated when the medium was tryptic soy broth (TSB), whereas the formation of terpineols was clearly favoured in malt broth (MB). Thus, there was a marked influence of the culture conditions on the results of the biotransformation. Changes in the conditions led to variations both in the type and relative amount of products obtained. PMID:24772824

García-Carnelli, Carlos; Rodríguez, Paula; Heinzen, Horacio; Menéndez, Pilar

2014-01-01

188

Morphology of Filamentous Fungi: Linking Cellular Biology to Process Engineering Using Aspergillus niger  

NASA Astrophysics Data System (ADS)

In various biotechnological processes, filamentous fungi, e.g. Aspergillus niger, are widely applied for the production of high value-added products due to their secretion efficiency. There is, however, a tangled relationship between the morphology of these microorganisms, the transport phenomena and the related productivity. The morphological characteristics vary between freely dispersed mycelia and distinct pellets of aggregated biomass. Hence, advantages and disadvantages for mycel or pellet cultivation have to be balanced out carefully. Due to this inadequate understanding of morphogenesis of filamentous microorganisms, fungal morphology, along with reproducibility of inocula of the same quality, is often a bottleneck of productivity in industrial production. To obtain an optimisation of the production process it is of great importance to gain a better understanding of the molecular and cell biology of these microorganisms as well as the approaches in biochemical engineering and particle technique, in particular to characterise the interactions between the growth conditions, cell morphology, spore-hyphae-interactions and product formation. Advances in particle and image analysis techniques as well as micromechanical devices and their applications to fungal cultivations have made available quantitative morphological data on filamentous cells. This chapter provides the ambitious aspects of this line of action, focussing on the control and characterisation of the morphology, the transport gradients and the approaches to understand the metabolism of filamentous fungi. Based on these data, bottlenecks in the morphogenesis of A. niger within the complex production pathways from gene to product should be identified and this may improve the production yield.

Krull, Rainer; Cordes, Christiana; Horn, Harald; Kampen, Ingo; Kwade, Arno; Neu, Thomas R.; Nörtemann, Bernd

189

Switching from a Unicellular to Multicellular Organization in an Aspergillus niger Hypha  

PubMed Central

ABSTRACT Pores in fungal septa enable cytoplasmic streaming between hyphae and their compartments. Consequently, the mycelium can be considered unicellular. However, we show here that Woronin bodies close ~50% of the three most apical septa of growing hyphae of Aspergillus niger. The incidence of closure of the 9th and 10th septa was even ?94%. Intercompartmental streaming of photoactivatable green fluorescent protein (PA-GFP) was not observed when the septa were closed, but open septa acted as a barrier, reducing the mobility rate of PA-GFP ~500 times. This mobility rate decreased with increasing septal age and under stress conditions, likely reflecting a regulatory mechanism affecting septal pore diameter. Modeling revealed that such regulation offers effective control of compound concentration between compartments. Modeling also showed that the incidence of septal closure in A. niger had an even stronger impact on cytoplasmic continuity. Cytoplasm of hyphal compartments was shown not to be in physical contact when separated by more than 4 septa. Together, data show that apical compartments of growing hyphae behave unicellularly, while older compartments have a multicellular organization. PMID:25736883

Bleichrodt, Robert-Jan; Hulsman, Marc

2015-01-01

190

Mutagenesis and genotypic characterization of Aspergillus niger FCBP-02 for improvement in cellulolytic potential.  

PubMed

Cellulase is a collective term that encompasses enzymes which catalyze reactions that participate in the degradation of insoluble cellulose to soluble carbohydrates. In the present study, production of extra cellular cellulases by a filamentous fungus, Aspergillus niger FCBP-02, was studied in solid-state fermentation (SSF) as well as in submerged fermentation (SmF). Trials were conducted to evaluate the effect of mutagenesis by UV irradiation (5-40 min) and ethyl methane sulfonate (EMS) treatment (50-300 microg mL(-1)) to obtain hyper active cellulase enzyme producers among the potential strains. The enzyme activity assays of parental and mutant strains clearly revealed significantly higher cellulase activity of mutant A-Ch-5.5 (96 Units mL(-1)), followed by A-UV-5.6 (71 Units mL(-1)) with respect to the wild strain of A. niger FCBP-02 (53.7 Units mL(-1)). The profile of genetic variability among wild and mutant derivatives was scrutinized through RAPD-PCR. The expression pattern of mutants exhibited that the mutants were isogenic variants of the wild type and the out performance of the mutants could be attributed to the change in genetic make up. PMID:19476005

Shafique, Shazia; Bajwa, Rukhsana; Shafique, Sobiya

2009-04-01

191

Heterologous Expression of Aspergillus niger ?-d-Xylosidase (XlnD): Characterization on Lignocellulosic Substrates  

NASA Astrophysics Data System (ADS)

The gene encoding a glycosyl hydrolase family 3 xylan 1,4-beta-xylosidase, xlnD, was successfully cloned from Aspergillus niger strain ATCC 10864. The recombinant product was expressed in Aspergillus awamori, purified by column chromatography, and verified by matrix-assisted laser desorption ionization, tandem time of flight (MALDI-TOF/ TOF) mass spectroscopy of tryptic digests. The T max was determined using differential scanning microcalorimetry (DSC) to be 78.2 °C; the K m and k cat were found to be 255 ?M and 13.7 s-1, respectively, using pNP-?-d-xylopyranoside as substrate. End-product inhibition by d-xylose was also verified and shown to be competitive; the K i for this inhibition was estimated to be 3.3 mM. XlnD was shown to efficiently hydrolyze small xylo-oligomers to monomeric xylose, making it a critical hydrolytic activity in cases where xylose is to be recovered from biomass conversion processes. In addition, the presence of the XlnD was shown to synergistically enhance the ability of an endoxylanase, XynA from Thermomyces lanuginosus, to convert xylan present in selected pretreated lignocellulosic substrates. Furthermore, the addition of the XynA/XlnD complex was effective in enhancing the ability of a simplified cellulase complex to convert glucan present in the substrates.

Selig, Michael J.; Knoshaug, Eric P.; Decker, Stephen R.; Baker, John O.; Himmel, Michael E.; Adney, William S.

192

Heterologous Expression of Aspergillus Niger --beta--D-Xylosidase (XInD): Characterization on Lignocellulosic Substrates  

SciTech Connect

The gene encoding a glycosyl hydrolase family 3 xylan 1,4-beta-xylosidase, xlnD, was successfully cloned from Aspergillus niger strain ATCC 10864. The recombinant product was expressed in Aspergillus awamori, purified by column chromatography, and verified by matrix-assisted laser desorption ionization, tandem time of flight (MALDI-TOF/TOF) mass spectroscopy of tryptic digests. The T{sub max} was determined using differential scanning microcalorimetry (DSC) to be 78.2 C; the K{sub m} and k{sub cat} were found to be 255 {micro}M and 13.7 s{sup -1}, respectively, using {rho}NP-{Beta}-d-xylopyranoside as substrate. End-product inhibition by d-xylose was also verified and shown to be competitive; the K{sub i} for this inhibition was estimated to be 3.3 mM. XlnD was shown to efficiently hydrolyze small xylo-oligomers to monomeric xylose, making it a critical hydrolytic activity in cases where xylose is to be recovered from biomass conversion processes. In addition, the presence of the XlnD was shown to synergistically enhance the ability of an endoxylanase, XynA from Thermomyces lanuginosus, to convert xylan present in selected pretreated lignocellulosic substrates. Furthermore, the addition of the XynA/XlnD complex was effective in enhancing the ability of a simplified cellulase complex to convert glucan present in the substrates.

Selig, M. J.; Knoshaug, E. P.; Decker, S. R.; Baker, J. O.; Himmel, M. E.; Adney, W. S.

2008-01-01

193

Biochemical properties of Cu/Zn-superoxide dismutase from fungal strain Aspergillus niger 26  

NASA Astrophysics Data System (ADS)

The fungal strain Aspergillus niger produces two superoxide dismutases, Cu/Zn-SOD and Mn-SOD. The primary structure of the Cu/Zn-SOD has been determined by Edman degradation of peptide fragments derived from proteolytic digests. A single chain of the protein, consisting of 153 amino acid residues, reveals a very high degree of structural homology with the amino acid sequences of other Aspergillus Cu/Zn-SODs. The molecular mass of ANSOD, measured by MALDI-MS and ESI-MS, and calculated by its amino acid sequence, was determined to be 15 821 Da. Only one Trp residue, at position 32, and one disulfide bridge were identified. However, neither a Tyr residue nor a carbohydrate chain occupying an N-linkage site (-Asn-Ile-Thr-) were found. Studies on the temperature and pH dependence of fluorescence, and on the temperature dependence of CD spectroscopic properties, confirmed that the enzyme is very stable, which can be explained by the stabilising effect of the disulfide bridge. The enzyme retains about 53% of its activity after incubation for a period of 30 min at 60 °C, and 15% at 85 °C.

Dolashki, Aleksandar; Abrashev, Radoslav; Stevanovic, Stefan; Stefanova, Lilyana; Ali, Syed Abid; Velkova, Ludmila; Hristova, Rumyana; Angelova, Maria; Voelter, Wolfgang; Devreese, Bart; Van Beeumen, Jozef; Dolashka-Angelova, Pavlina

2008-12-01

194

Genomic analysis of the aconidial and high-performance protein producer, industrially relevant Aspergillus niger SH2 strain.  

PubMed

Aspergillus niger is usually regarded as a beneficial species widely used in biotechnological industry. Obtaining the genome sequence of the widely used aconidial A. niger SH2 strain is of great importance to understand its unusual production capability. In this study we assembled a high-quality genome sequence of A. niger SH2 with approximately 11,517 ORFs. Relatively high proportion of genes enriched for protein expression related FunCat items verify its efficient capacity in protein production. Furthermore, genome-wide comparative analysis between A. niger SH2 and CBS513.88 reveals insights into unique properties of A. niger SH2. A. niger SH2 lacks the gene related with the initiation of asexual sporulation (PrpA), leading to its distinct aconidial phenotype. Frame shift mutations and non-synonymous SNPs in genes of cell wall integrity signaling, ?-1,3-glucan synthesis and chitin synthesis influence its cell wall development which is important for its hyphal fragmentation during industrial high-efficiency protein production. PMID:24630962

Yin, Chao; Wang, Bin; He, Pan; Lin, Ying; Pan, Li

2014-05-15

195

Citric acid production by solid-state fermentation in a packed-bed reactor using Aspergillus niger  

Microsoft Academic Search

Kumara, a starch-containing root crop grown extensively in New Zealand, has been used as a substrate for citric acid production using Aspergillus niger in solid-state fermentation. When the process was operated in a packed-bed reactor, the bed loading was the most important operational parameter. The airflow rate and substrate particle size were also important but their net effects varied depending

Minyuan Lu; John D. Brooks; Ian S. Maddox

1997-01-01

196

Removal of arsenic from aqueous environments by native and chemically modified biomass of Aspergillus niger and Neosartorya fischeri  

Microsoft Academic Search

Arsenic removal from aqueous solutions by biomass of two fungal strains, Aspergillus niger and Neosartorya fischeri, was assessed. The biosorption capacity of fungal biomass was studied within the As(V) concentration range of approximately 0.2 to 5.0 mg L at two different pH values (pH 5 and 7). With increasing initial arsenic concentration, the biosorption capacity of both fungal strains increased

Pavol Littera; Martin Urík; Jaroslav Ševc; Marek Kolen?ík; Katarína Gardošová; Marianna Molnárová

2011-01-01

197

Differential expression of three alpha-galactosidase genes and a single beta-galactosidase gene from Aspergillus niger  

Microsoft Academic Search

A gene encoding a third a-galactosidase (AglB) from Aspergillus niger has been cloned and sequenced. The gene consists of an open reading frame of 1,750 bp containing six introns. The gene encodes a protein of 443 amino acids which contains a eukaryotic signal sequence of 16 amino acids and seven putative N-glycosylation sites. The mature protein has a calculated molecular

Vries de R. P; Broeck van den H. C; ESTER DEKKERS; PALOMA MANZANARES; Graaff de L. H; JAAP VISSER

1999-01-01

198

Release of ferulic acid from wheat bran by a ferulic acid esterase (FAE-III) from Aspergillus niger  

Microsoft Academic Search

Ferulic acid was efficiently released from a wheat bran preparation by a ferulic acid esterase from Aspergillus niger (FAE-III) when incubated together with a Trichoderma viride xylanase (a maximum of 95% total ferulic acid released after 5 h incubation). FAE-III by itself could release ferulic acid but at a level almost 24-fold lower than that obtained in the presence of

C. B. Faulds; G. Williamson

1995-01-01

199

Genetic analysis and the construction of master strains for assignment of genes to six linkage groups in Aspergillus niger  

Microsoft Academic Search

A start has been made on establishing a collection of Aspergillus niger colour and auxotrophic mutants with an isogenic background for use as a source of genetic markers. All strains have short conidiophores (cspAl ), which makes them easy to handle on test plates. Genetic markers were combined stepwise by somatic recombination. Somatic diploids were obtained at frequencies of 10-6-10-5

C. J. Bos; A. J. M. Debets; K. Swart; A. Huybers; G. Kobus; S. M. Slakhorst

1988-01-01

200

Studies of In Vitro Activities of Voriconazole and Itraconazole against Aspergillus Hyphae Using Viability Staining  

Microsoft Academic Search

The minimal fungicidal concentrations (MFCs) of voriconazole and itraconazole for five clinical isolates each of Aspergillus terreus, Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger were determined by a broth macrodilution method. Conidial suspensions as inocula were compared to hyphae as inocula since the invasive form of aspergillosis is manifested by the appearance of hyphal structures. In addition, cell viability staining

CORNELIA LASS-FLORL; MARKUS NAGL; CORNELIA SPETH; HANNO ULMER; MANFRED P. DIERICH; REINHARD WURZNER

2001-01-01

201

Aspergillus niger ?-Glucosidase Has a Cellulase-like Tadpole Molecular Shape  

PubMed Central

Aspergillus niger is known to secrete large amounts of ?-glucosidases, which have a variety of biotechnological and industrial applications. Here, we purified an A. niger ?-glucosidase (AnBgl1) and conducted its biochemical and biophysical analyses. Purified enzyme with an apparent molecular mass of 116 kDa forms monomers in solution as judged by native gel electrophoresis and has a pI value of 4.55, as found for most of the fungi of ?-glucosidases. Surprisingly, the small angle x-ray experiments reveal that AnBgl1 has a tadpole-like structure, with the N-terminal catalytic domain and C-terminal fibronectin III-like domain (FnIII) connected by the long linker peptide (?100 amino acid residues) in an extended conformation. This molecular organization resembles the one adopted by other cellulases (such as cellobiohydrolases, for example) that frequently contain a catalytic domain linked to the cellulose-binding module that mediates their binding to insoluble and polymeric cellulose. The reasons why AnBgl1, which acts on the small soluble substrates, has a tadpole molecular shape are not entirely clear. However, our enzyme pulldown assays with different polymeric substrates suggest that AnBgl1 has little or no capacity to bind to and to adsorb cellulose, xylan, and starch, but it has high affinity to lignin. Molecular dynamics simulations suggested that clusters of residues located in the C-terminal FnIII domain interact strongly with lignin fragments. The simulations showed that numerous arginine residues scattered throughout the FnIII surface play an important role in the interaction with lignin by means of cation-? stacking with the lignin aromatic rings. These results indicate that the C-terminal FnIII domain could be operational for immobilization of the enzyme on the cell wall and for the prevention of unproductive binding of cellulase to the biomass lignin. PMID:24064212

Lima, Marisa A.; Oliveira-Neto, Mario; Kadowaki, Marco Antonio S.; Rosseto, Flavio R.; Prates, Erica T.; Squina, Fabio M.; Leme, Adriana F. P.; Skaf, Munir S.; Polikarpov, Igor

2013-01-01

202

Structural Features of Sugars That Trigger or Support Conidial Germination in the Filamentous Fungus Aspergillus niger  

PubMed Central

The asexual spores (conidia) of Aspergillus niger germinate to produce hyphae under appropriate conditions. Germination is initiated by conidial swelling and mobilization of internal carbon and energy stores, followed by polarization and emergence of a hyphal germ tube. The effects of different pyranose sugars, all analogues of d-glucose, on the germination of A. niger conidia were explored, and we define germination as the transition from a dormant conidium into a germling. Within germination, we distinguish two distinct stages, the initial swelling of the conidium and subsequent polarized growth. The stage of conidial swelling requires a germination trigger, which we define as a compound that is sensed by the conidium and which leads to catabolism of d-trehalose and isotropic growth. Sugars that triggered germination and outgrowth included d-glucose, d-mannose, and d-xylose. Sugars that triggered germination but did not support subsequent outgrowth included d-tagatose, d-lyxose, and 2-deoxy-d-glucose. Nontriggering sugars included d-galactose, l-glucose, and d-arabinose. Certain nontriggering sugars, including d-galactose, supported outgrowth if added in the presence of a complementary triggering sugar. This division of functions indicates that sugars are involved in two separate events in germination, triggering and subsequent outgrowth, and the structural features of sugars that support each, both, or none of these events are discussed. We also present data on the uptake of sugars during the germination process and discuss possible mechanisms of triggering in the absence of apparent sugar uptake during the initial swelling of conidia. PMID:23995938

Hayer, Kimran; Stratford, Malcolm

2013-01-01

203

Biochar enhances Aspergillus niger rock phosphate solubilization by increasing organic acid production and alleviating fluoride toxicity.  

PubMed

During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F(-)) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F(-) adsorbents [activated alumina (Al2O3) and biochar] on RP solubilization by Aspergillus niger was examined. Al2O3 adsorbed part of the F(-) released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F(-) while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F(-) measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F(-) per liter can be removed from solution by biochar when added at 3 g liter(-1) to the culture medium. Thus, biochar acted as an F(-) sink during RP solubilization and led to an F(-) concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F(-) and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F(-), the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP. PMID:24610849

Mendes, Gilberto de Oliveira; Zafra, David Lopez; Vassilev, Nikolay Bojkov; Silva, Ivo Ribeiro; Ribeiro, José Ivo; Costa, Maurício Dutra

2014-05-01

204

On the safety of a new generation of DSM Aspergillus niger enzyme production strains.  

PubMed

Consumers safety of enzyme preparations is determined by three variables: the producing organism, the raw materials used in the production, and the production process itself. The latter one is embedded in current Good Manufacturing Practice (cGMP) and Hazard Analysis of Critical Control Points (HACCP); therefore the safety focus can be directed to raw materials and the producing organism. In this paper, we describe the use of novel genetically modified strains of Aspergillus niger-made by a design and build strategy-from a lineage of classically improved strains with a history of safe use in enzyme production. The specifics of the host strain allow for integration and over-expression of any gene of interest at a targeted integration site implying that the rest of the host genome is not affected by this integration. Furthermore due to the fact that the newly integrated gene copies are put under the genetic regulation of the host's own glucoamylase promoter, the recipe of the production process of any new production strain can be kept constant with respect to the raw materials composition. Consequently the safety of a new enzyme product from these novel genetically modified strains is determined by the background of the production organism. The use of a strain with a history of safe use and targeted integration according to the concept described above has consequences for the safety studies on the final product. If a known enzymatic activity is over-expressed the safety of a new enzyme preparation is covered by the results of the safety studies performed for other strains from this specific Aspergillus niger strain lineage. In this paper an overview is given on the available toxicity tests with these strains. We conclude that for new enzyme products produced with strains from this lineage using the design and build technology no new sub-acute/chronic oral toxicity studies are needed. This also has the benefit that no longer test animals are needed to demonstrate the safety of products produced by these strains. PMID:12878051

van Dijck, Piet W M; Selten, Gerard C M; Hempenius, Rixta A

2003-08-01

205

Secretion and properties of a hybrid Kluyveromyces lactis-Aspergillus niger ?-galactosidase  

PubMed Central

Background The ?-galactosidase from Kluyveromyces lactis is a protein of outstanding biotechnological interest in the food industry and milk whey reutilization. However, due to its intracellular nature, its industrial production is limited by the high cost associated to extraction and downstream processing. The yeast-system is an attractive method for producing many heterologous proteins. The addition of a secretory signal in the recombinant protein is the method of choice to sort it out of the cell, although biotechnological success is not guaranteed. The cell wall acting as a molecular sieve to large molecules, culture conditions and structural determinants present in the protein, all have a decisive role in the overall process. Protein engineering, combining domains of related proteins, is an alternative to take into account when the task is difficult. In this work, we have constructed and analyzed two hybrid proteins from the ?-galactosidase of K. lactis, intracellular, and its Aspergillus niger homologue that is extracellular. In both, a heterologous signal peptide for secretion was also included at the N-terminus of the recombinant proteins. One of the hybrid proteins obtained has interesting properties for its biotechnological utilization. Results The highest levels of intracellular and extracellular ?-galactosidase were obtained when the segment corresponding to the five domain of K. lactis ?-galactosidase was replaced by the corresponding five domain of the A. niger ?-galactosidase. Taking into account that this replacement may affect other parameters related to the activity or the stability of the hybrid protein, a thoroughly study was performed. Both pH (6.5) and temperature (40°C) for optimum activity differ from values obtained with the native proteins. The stability was higher than the corresponding to the ?-galactosidase of K. lactis and, unlike this, the activity of the hybrid protein was increased by the presence of Ni2+. The affinity for synthetic (ONPG) or natural (lactose) substrates was higher in the hybrid than in the native K. lactis ?-galactosidase. Finally, a structural-model of the hybrid protein was obtained by homology modelling and the experimentally determined properties of the protein were discussed in relation to it. Conclusion A hybrid protein between K. lactis and A. niger ?-galactosidases was constructed that increases the yield of the protein released to the growth medium. Modifications introduced in the construction, besides to improve secretion, conferred to the protein biochemical characteristics of biotechnological interest. PMID:17176477

Rodríguez, Ángel Pereira; Leiro, Rafael Fernández; Trillo, M Cristina; Cerdán, M Esperanza; Siso, M Isabel González; Becerra, Manuel

2006-01-01

206

Aspergillus pragensis sp. nov. discovered during molecular reidentification of clinical isolates belonging Aspergillus section Candidi  

Technology Transfer Automated Retrieval System (TEKTRAN)

The identity of nine clinical isolates from Czech patients presumably belonging to Aspergillus section Candidi based on morphology of colonies was revised using sequences of ß-tubulin, calmodulin, and internal transcribed spacer (ITS) rDNA. The set of isolates included six isolates from suspected (n...

207

Purification and properties of one component of acid phosphatase produced by Aspergillus niger.  

PubMed

One component, the i form, of acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) produced by Aspergillus niger was purified from the mycelial extract. The purified enzyme was homogenous on Sephadex G-200 gel filtration, disc electrophoresis and heat inactivation. The purified enzyme was studied and the following results were obtained: 1. The enzyme catalyzed the hydrolysis of a wide variety of phosphomonoesters, but not that of bis(p-nitrophenyl)phosphate, adenosine 3',5'-cyclic monophosphate, fructose 1,6-diphosphate, adenosine 5'-diphosphate or adenosine 5'-triphosphate. 2. Fluoride, orthophosphate, arsenate, borate, molybdate and (+)-tartrate acted as inhibitors. This enzyme was inactivated by N-bromosuccinimide and 2-hydroxy-5-nitrobenzyl bromide, and was not affected by p-chloromercuribenzoate, N-acetylimidazole, p-diazobenzenesulfonic acid and tetranitromethane. From these results, tryptophan was estimated to play an important role in the enzyme activity. 3. The apparent molecular weight was 310000 by Sephadex G-200 gel filtration. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate suggested that the molecular weight of the subunit was approximately 89000. 4. The purified enzyme contained 29% carbohydrate consisting of glucosamine, mannose and galactose. The amino acid composition of this enzyme was not specific compared with other known acid phosphatases. PMID:13843

Shimada, Y; Shinmyo, A; Enatsu, T

1977-02-01

208

Effect of Aspergillus niger xylanase on dough characteristics and bread quality attributes.  

PubMed

The present study was conducted to investigate the impact of various treatments of xylanase produced by Aspergillus niger applied in bread making processes like during tempering of wheat kernels and dough mixing on the dough quality characteristics i.e. dryness, stiffness, elasticity, extensibility, coherency and bread quality parameters i.e. volume, specific volume, density, moisture retention and sensory attributes. Different doses (200, 400, 600, 800 and 1,000 IU) of purified enzyme were applied to 1 kg of wheat grains during tempering and 1 kg of flour (straight grade flour) during mixing of dough in parallel. The samples of wheat kernels were agitated at different intervals for uniformity in tempering. After milling and dough making of both types of flour (having enzyme treatment during tempering and flour mixing) showed improved dough characteristics but the improvement was more prominent in the samples receiving enzyme treatment during tempering. Moreover, xylanase decreased dryness and stiffness of the dough whereas, resulted in increased elasticity, extensibility and coherency and increase in volume & decrease in bread density. Xylanase treatments also resulted in higher moisture retention and improvement of sensory attributes of bread. From the results, it is concluded that dough characteristics and bread quality improved significantly in response to enzyme treatments during tempering as compared to application during mixing. PMID:25328183

Ahmad, Zulfiqar; Butt, Masood Sadiq; Ahmed, Anwaar; Riaz, Muhammad; Sabir, Syed Mubashar; Farooq, Umar; Rehman, Fazal Ur

2014-10-01

209

Optimization of xylanase production from Aspergillus niger for biobleaching of eucalyptus pulp.  

PubMed

A crude endo-xylanase produced by Aspergillus niger BCC14405 was investigated for its potential in pre-bleaching of chemical pulp from eucalyptus. The optimal fermentation conditions on the basis of optimization using response surface methodology included cultivation in a complex medium comprising wheat bran, rice bran, and soybean meal supplemented with yeast extract, glucose, peptone, and lactose with a starting pH of 6.0 for 7 d. This resulted in production of 89.5 IU/mL of xylanase with minor cellulase activity. Proteomic analysis using LC/MS/MS revealed that the crude enzyme was a composite of hemicellulolytic enzymes, including endo-?-1,4-xylanase and other hemicellulolytic enzymes attacking arabinoxylan and mannan. Pretreatment of the pulp at a xylanase dosage of 10 IU/g increased the brightness ceiling after the C-Eop-H bleaching step up to 3.0% using a chlorine charge with a C-factor of 0.16-0.20. Xylanase treatment also led to reduction in chlorine charge of at least 20%, with an acceptable brightness level. The enzyme pretreatment resulted in a slight increase in pulp viscosity, suggesting an increase in relative cellulose content. The crude enzyme was potent in the enzyme-aided bleaching of chemical pulp in an environmentally friendly pulping process. PMID:21670524

Khonzue, Parichart; Laothanachareon, Thanaporn; Rattanaphan, Nakul; Tinnasulanon, Phungjai; Apawasin, Saowanee; Paemanee, Atchara; Ruanglek, Vasimon; Tanapongpipat, Sutipa; Champreda, Verawat; Eurwilaichitr, Lily

2011-01-01

210

Sorption of heavy metals by the soil fungi 'Aspergillus niger' and Mucor rouxii  

SciTech Connect

Sorption of the nitrate salts of cadmium(II), copper(II), lanthanum(III) and silver(I) by two fungi, Aspergillus niger and Mucor rouxii, was evaluated using Freundlich adsorption isotherms and energy dispersive X-ray electron microscopy. The linearized Freundlich isotherm described the metal sorption data well for metal concentrations of 5 microM-1 mM metal. Differences in metal binding were observed among metals, as well as between fungal species. Calculated Freundlich K values indicated that metal binding decreased in the order La(3+) > or = Ag(+) > Cu(2+) > Cd(2+). However, sorption of Ag(+) was greater than that of La(3+) from solutions of 0.1 and 1 mM metal and likely due to precipitation at the cell wall surface. At the 1 mM initial concentration, there were no significant differences between the two fungi in metal sorption, except for Ag(+) binding. At the 5 microM concentration, there was no difference between the fungi in their sorption capacities for the four metals. Electron microscopy-energy dispersive X-ray analysis indicated that silver precipitated onto cells as colloidal silver. The results indicate that Freundlich isotherms may be useful for describing short-term metal sorption by fungal biomass and for comparison with other soil constituents in standardized systems. (Copyright (c) 1992 Pergamon Press plc.)

Mullen, M.D.; Wolf, D.C.; Beveridge, T.J.; Bailey, G.W.

1992-01-01

211

The active site topology of Aspergillus niger endopolygalacturonase II as studied by site-directed mutagenesis.  

PubMed

Strictly conserved charged residues among polygalacturonases (Asp-180, Asp-201, Asp-202, His-223, Arg-256, and Lys-258) were subjected to site-directed mutagenesis in Aspergillus niger endopolygalacturonase II. Specific activity, product progression, and kinetic parameters (K(m) and V(max)) were determined on polygalacturonic acid for the purified mutated enzymes, and bond cleavage frequencies on oligogalacturonates were calculated. Depending on their specific activity, the mutated endopolygalacturonases II were grouped into three classes. The mutant enzymes displayed bond cleavage frequencies on penta- and/or hexagalacturonate different from the wild type endopolygalacturonase II. Based on the biochemical characterization of endopolygalacturonase II mutants together with the three-dimensional structure of the wild type enzyme, we suggest that the mutated residues are involved in either primarily substrate binding (Arg-256 and Lys-258) or maintaining the proper ionization state of a catalytic residue (His-223). The individual roles of Asp-180, Asp-201, and Asp-202 in catalysis are discussed. The active site topology is different from the one commonly found in inverting glycosyl hydrolases. PMID:10617668

Armand, S; Wagemaker, M J; Sánchez-Torres, P; Kester, H C; van Santen, Y; Dijkstra, B W; Visser, J; Benen, J A

2000-01-01

212

Aspergillus Niger peritonitis in a peritoneal dialysis patient treated with eculizumab.  

PubMed

The complement system plays a vital role in preventing life-threatening infections by ensuring optimal functioning of the host immune system. Its dysregulation has been implicated in causing glomerular, hematological, and transplant-related disorders. Eculizumab a novel monoclonal antibody against complement component C5 has emerged in the recent past as the standard of care offering an effective rescue and maintenance therapy against many of these disorders. Its use has been associated with increased risk of infections predominantly with encapsulated organisms. There is no data in the literature on its effects in end-stage kidney disease (ESKD) or dialysis patients. We describe here a very rare case of Aspergillus Niger peritonitis in an ESKD patient on peritoneal dialysis (PD) receiving maintenance eculizumab therapy for atypical hemolytic uremic syndrome. Given that murine models with the same defect as that induced by eculizumab is vulnerable to invasive Aspergillosis, it is suggested that the fungal peritonitis in this patient was the result of the eculizumab therapy. PMID:24512095

Vellanki, Venkat S; Bargman, Joanne M

2014-05-01

213

Response surface optimization of fermentation conditions for producing xylanase by Aspergillus niger SL-05.  

PubMed

Fermentation conditions were statistically optimized for producing extracellular xylanase by Aspergillus niger SL-05 using apple pomace and cotton seed meal. The primary study shows that culture medium with a 1:1 ratio of apple pomace and cotton seed meal (carbon and nitrogen sources) yielded maximal xylanase activity. Three significant factors influencing xylanase production were identified as urea, KH(2)PO(4), and initial moisture content using Plackett-Burman design study. The effects of these three factors were further investigated using a design of rotation-regression-orthogonal combination. The optimized conditions by response surface analysis were 2.5% Urea, 0.09% KH(2)PO(4), and 62% initial moisture content. The analysis of variance indicated that the established model was significant (P < 0.05), "while" or "and" the lack of fit was not significant. Under the optimized conditions, the model predicted 4,998 IU/g dry content, whereas validation experiments produced an enzymatic activity of xylanase at 5,662 IU/g dry content after 60 h fermentation. This study innovatively developed a fermentation medium and process to utilize inexpensive agro-industrial wastes to produce a high yield of xylanase. PMID:18309527

Liu, Cheng; Sun, Zhong-Tao; Du, Jin-Hua; Wang, Jian

2008-07-01

214

Bioleaching of nickel and cobalt from lateritic chromite overburden using the culture filtrate of Aspergillus niger.  

PubMed

Extraction of metals (Ni, Co) from chromite overburden of Sukinda mines of Orissa, India, with the culture filtrate of Aspergillus niger was studied. Results showed that the amounts of metals leached varied directly with reaction temperature and period of fermentation. The culture filtrate was analyzed for citric and oxalic acids, and contained only oxalic acid-the concentration of which increased with time. Although this acid played the major role in leaching of metals, other unidentified metabolites present in the culture filtrate influenced the dissolution of the metals significantly. Maximum recovery of metals from raw and roasted ore samples was achieved at 80 °C with the 21-day culture filtrate containing the highest amount of oxalic acid. Under identical experimental conditions, much higher amounts of the metals were leached from roasted ore. Microstructures of the ore particles were studied by scanning electron microscopy and transmission electron microscopy; the bonding behaviors of metal compounds were identified by Fourier transform infrared spectroscopy which showed that the metals were leached after chelation with oxalic acid. PMID:23700146

Biswas, Supratim; Dey, Rajib; Mukherjee, Siddhartha; Banerjee, Pataki C

2013-08-01

215

The effect of natamycin on the transcriptome of conidia of Aspergillus niger.  

PubMed

The impact of natamycin on Aspergillus niger was analysed during the first 8 h of germination of conidia. Polarisation, germ tube formation, and mitosis were inhibited in the presence of 3 and 10 ?M of the anti-fungal compound, while at 10 ?M also isotropic growth was affected. Natamycin did not have an effect on the decrease of microviscosity during germination and the concomitant reduction in mannitol and trehalose levels. However, it did abolish the increase of intracellular levels of glycerol and glucose during the 8 h period of germination.Natamycin hardly affected the changes that occur in the RNA profile during the first 2 h of germination. During this time period, genes related to transcription, protein synthesis, energy and cell cycle and DNA processing were particularly up-regulated. Differential expression of 280 and 2586 genes was observed when 8 h old germlings were compared with conidia that had been exposed to 3 ?M and 10 ?M natamycin, respectively. For instance, genes involved in ergosterol biosynthesis were down-regulated. On the other hand, genes involved in endocytosis and the metabolism of compatible solutes, and genes encoding protective proteins were up-regulated in natamycin treated conidia. PMID:23449730

van Leeuwen, M R; Krijgsheld, P; Wyatt, T T; Golovina, E A; Menke, H; Dekker, A; Stark, J; Stam, H; Bleichrodt, R; Wösten, H A B; Dijksterhuis, J

2013-03-15

216

Antifungal agents against Aspergillus niger for rearing rice leaffolder larvae (Lepidoptera: Pyralidae) on artificial diet.  

PubMed

Mold contamination is an important issue in insect mass rearing. Frequently used antifungal agents such as sorbic acid and methylparaben have negative impact on many lepidopteran larvae, which might be one of the reasons for the difficulty in rearing rice leaffolder, Cnaphalocrocis medinalis (Güenée). In this study, 19 antifungal agents, including 7 food preservatives, 6 antifungal drugs, and 6 agricultural fungicides, were screened for their inhibitory activities on Aspergillus niger in diets. The results demonstrated that most of the tested chemicals are unsuitable as mold inhibitors in the diets of the rice leaffolder, and the rice leaffolder neonate is sensitive to sorbic acid and methylparaben. These two mold inhibitors at commonly used concentrations were shown to impact the survival of rice leaffolder larvae fed on artificial diets. Among the tested mold inhibitors, natamycin was the safest for the rice leaffolder larvae. Much higher larva survival was observed for the larvae fed on diets containing natamycin as an antifungal agent (59 and 72% at 200 and 400 ppm, respectively). Two agricultural fungicides, tebuconazole and azoxystrobin, are also potent as mold inhibitors when used in insect diets. The mixed use of natamycin and sorbic acid, or methylparaben, and the mixed use of sorbic acid and azoxystrobin resulted in significantly higher larva survival than sorbic acid + methylparaben. Natamycin + azoxystrobin and sorbic acid + tebuconazole resulted in larva survival similar to that of sorbic acid + methylparaben. The ternary combination of natamycin, sorbic acid, and methylparaben was the best combination for the rearing of rice leaffolder. PMID:25026669

Su, Jianya; Wang, Ye-Cheng; Zhang, Shu-Kun; Ren, Xiu-Bei

2014-06-01

217

Fluoride-Tolerant Mutants of Aspergillus niger Show Enhanced Phosphate Solubilization Capacity  

PubMed Central

P-solubilizing microorganisms are a promising alternative for a sustainable use of P against a backdrop of depletion of high-grade rock phosphates (RPs). Nevertheless, toxic elements present in RPs, such as fluorine, can negatively affect microbial solubilization. Thus, this study aimed at selecting Aspergillus niger mutants efficient at P solubilization in the presence of fluoride (F?). The mutants were obtained by exposition of conidia to UV light followed by screening in a medium supplemented with Ca3(PO4)2 and F?. The mutant FS1-555 showed the highest solubilization in the presence of F?, releasing approximately 70% of the P contained in Ca3(PO4)2, a value 1.7 times higher than that obtained for the wild type (WT). The mutant FS1-331 showed improved ability of solubilizing fluorapatites, increasing the solubilization of Araxá, Catalão, and Patos RPs by 1.7, 1.6, and 2.5 times that of the WT, respectively. These mutants also grew better in the presence of F?, indicating that mutagenesis allowed the acquisition of F? tolerance. Higher production of oxalic acid by FS1-331 correlated with its improved capacity for RP solubilization. This mutant represents a significant improvement and possess a high potential for application in solubilization systems with fluoride-rich phosphate sources. PMID:25310310

Silva, Ubiana de Cássia; Mendes, Gilberto de Oliveira; Silva, Nina Morena R. M.; Duarte, Josiane Leal; Silva, Ivo Ribeiro; Tótola, Marcos Rogério; Costa, Maurício Dutra

2014-01-01

218

Exploring sequence characteristics related to high-level production of secreted proteins in Aspergillus niger.  

PubMed

Protein sequence features are explored in relation to the production of over-expressed extracellular proteins by fungi. Knowledge on features influencing protein production and secretion could be employed to improve enzyme production levels in industrial bioprocesses via protein engineering. A large set, over 600 homologous and nearly 2,000 heterologous fungal genes, were overexpressed in Aspergillus niger using a standardized expression cassette and scored for high versus no production. Subsequently, sequence-based machine learning techniques were applied for identifying relevant DNA and protein sequence features. The amino-acid composition of the protein sequence was found to be most predictive and interpretation revealed that, for both homologous and heterologous gene expression, the same features are important: tyrosine and asparagine composition was found to have a positive correlation with high-level production, whereas for unsuccessful production, contributions were found for methionine and lysine composition. The predictor is available online at http://bioinformatics.tudelft.nl/hipsec. Subsequent work aims at validating these findings by protein engineering as a method for increasing expression levels per gene copy. PMID:23049690

van den Berg, Bastiaan A; Reinders, Marcel J T; Hulsman, Marc; Wu, Liang; Pel, Herman J; Roubos, Johannes A; de Ridder, Dick

2012-01-01

219

Influence of dietary components on Aspergillus niger prolyl endoprotease mediated gluten degradation.  

PubMed

Celiac disease (CD) is caused by intolerance to gluten. Oral supplementation with enzymes like Aspergillus niger propyl-endoprotease (AN-PEP), which can hydrolyse gluten, has been proposed to prevent the harmful effects of ingestion of gluten. The influence of meal composition on AN-PEP activity was investigated using an in vitro model that simulates stomach-like conditions. AN-PEP optimal dosage was 20 proline protease units (PPU)/g gluten. The addition of a carbonated drink strongly enhanced AN-PEP activity because of its acidifying effect. While fat did not affect gluten degradation by AN-PEP, the presence of food proteins slowed down gluten detoxification. Moreover, raw gluten was degraded more efficiently by AN-PEP than baked gluten. We conclude that the meal composition influences the amount of AN-PEP needed for gluten elimination. Therefore, AN-PEP should not be used to replace a gluten free diet, but rather to support digestion of occasional and/or inadvertent gluten consumption. PMID:25529703

Montserrat, Veronica; Bruins, Maaike J; Edens, Luppo; Koning, Frits

2015-05-01

220

Purification and Characterization of a Ginsenoside Rb1-Hydrolyzing ?-Glucosidase from Aspergillus niger KCCM 11239  

PubMed Central

Rb1-hydrolyzing ?-glucosidase from Aspergillus niger KCCM 11239 was studied to develop a bioconversion process for minor ginsenosides. The specific activity of the purified enzyme was 46.5 times greater than that of the crude enzyme. The molecular weight of the native enzyme was estimated to be approximately 123 kDa. The optimal pH of the purified enzyme was pH 4.0, and the enzyme proved highly stable over a pH range of 5.0–10.0. The optimal temperature was 70 °C, and the enzyme became unstable at temperatures above 60 °C. The enzyme was inhibited by Cu2+, Mg2+, Co2+, and acetic acid (10 mM). In the specificity tests, the enzyme was found to be active against ginsenoside Rb1, but showed very low levels of activity against Rb2, Rc, Rd, Re, and Rg1. The enzyme hydrolyzed the 20-C,?-(1?6)-glucoside of ginsenoside Rb1 to generate ginsenoside Rd and Rg3, and hydrolyzed 3-C,?-(1?2)-glucoside to generate F2. The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds. PMID:23109906

Chang, Kyung Hoon; Jo, Mi Na; Kim, Kee-Tae; Paik, Hyun-Dong

2012-01-01

221

Properties of a ?-(1?4)-glucan hydrolase from Aspergillus niger  

PubMed Central

1. A ?-(1?4)-glucan hydrolase prepared from Aspergillus niger, as described by Clarke & Stone (1965a), showed a pH optimum in the range 4·5–6 and Km 0·25% when acting on a cellulose dextrin sulphate substrate. 2. The hydrolase rapidly decreased the specific viscosity of carboxymethylcellulose with a small increase in the production of reducing sugars. The identity of the products of hydrolysis of cellotetraose, cellopentaose and their reduced analogues indicate a preferential cleavage of non-terminal glucosidic linkages. The enzyme may be described as ?-(1?4)-glucan 4-glucanohydrolase (EC 3.2.1.4). 3. In addition to carboxymethylcellulose, cellulose dextrins, cellopentaose and cellotetraose the enzyme fraction hydrolysed lichenin, oat and barley glucans, ivory-nut mannan and a glucomannan from Konjak flour. No hydrolysis of wheat-straw ?-(1?4)-xylan, Lupinus albus ?-(1?4)-galactan, pneumococcal type III polysaccharide, chitin, hyaluronic acid, laminarin, pachydextrins, carboxymethylpachyman or ?-(1?3)-oligoglucosides was detected. 4. The hydrolase showed no transglycosylase activity from cellodextrin or cellopentaose substrates to glucose or methanol acceptors. 5. The hydrolysis of cellodextrins was inhibited completely by 1·0mm-Hg2+, 0·7mm-phenylmercuric nitrate and 1·0mm-iodine. PMID:5862418

Clarke, A. E.; Stone, B. A.

1965-01-01

222

Niger.  

PubMed

Niger is two-thirds Sahara desert and the rest savannah with an area irrigated by the Niger River valley. The 6.2 million people are therefore either nomadic herdsmen or subsistence farmers, coping with a hot, dry climate. There are 5 or more ethnic groups, 2 main languages other than the official French, and most people are Muslim. The growth rate is 3.1%; children make up 45% of the population; infant mortality is 145/1000; life expectancy is 44.5 years. The constitutional government has been suspended by a military regime. A multi-layered structure called "development society" has been instituted. Per capita income is about $265. Niger has uranium, coal, iron, tin and phosphates, and farm products include peanuts, millet, sorghum, beans, cotton, rice and cowpeas. Niger received assistance from France, US, West Germany, Canada, Saudi Arabia, as well as international organizations and military assistance from several countries. PMID:12177951

1987-06-01

223

Phytotoxicity of lignanamides isolated from the seeds of Hyoscyamus niger.  

PubMed

Bioassay-guided fractionation of phytotoxic extracts prepared from the seeds of Hyoscyamus niger led to the isolation of three new lignanamides (1-3), along with six known lignanamides (4-9). The structures of the new compounds were determined by spectroscopic methods, including 1D and 2D nuclear magnetic resonance techniques, and high-resolution electrospray ionization mass spectrometry. The bioactivity analysis of the isolated compounds showed that compound 3 exhibited significant inhibition on the germination and radical elongation of Allium fistulosum at 10(-4) M concentration. PMID:22280058

Zhang, Wen-Na; Luo, Jian-Guang; Kong, Ling-Yi

2012-02-22

224

Effects of cellulase from Aspergillus niger and solvent pretreatments on the extractability of organic green tea waste.  

PubMed

As green tea is being consumed in larger amounts, more green tea waste is being produced. Following extraction, several bioactive compounds may exist in the waste including polyphenols and amino acids. It was found that an Aspergillus niger cellulase treatment of green tea waste increased the extractability of various nutritional and functional components after pretreatments with various extraction solvents such as cold water (CW), hot water (HW), sulfuric acid (SA), hydrochloric acid (HA), and methanol (Me). After the residue was treated with cellulase from Aspergillus niger, the amounts of polyphenols, total catechins, and reducing sugars in the HW extract were increased by 64.6, 941.2, and 350.9%, respectively. In particular, levels of epigallocatechin, epicatechin, and gallic acid were significantly enhanced compared to those in the nontreated control. However, protein extraction was not significantly affected, and cellulase treatment was not more efficient for caffeine extraction compared to phenolic extraction. Among the four extraction solvents, HW and SA showed relatively higher extractabilities as compared to the other groups (CW, HA, and Me). These results indicate that cellulase from A. niger can increase the extractability of green tea waste when combined with certain solvent pretreatments. Consequently, the residual functional compounds and essential nutrients from cellulase-treated green tea waste have the potential to be applied in the production of new functional foods. PMID:20843026

Kim, Jae Hun; Pan, Jeong Hoon; Heo, Wan; Lee, Hyungjae; Kwon, Eung Gi; Lee, Hong-Gu; Shin, Dong Hoon; Liu, Rui Hai; Kim, Young Jun

2010-10-13

225

Utilization of molasses and sugar cane bagasse for production of fungal invertase in solid state fermentation using Aspergillus niger GH1  

PubMed Central

Agro-industrial wastes have been used as substrate-support in solid state fermentation for enzyme production. Molasses and sugarcane bagasse are by-products of sugar industry and can be employed as substrates for invertase production. Invertase is an important enzyme for sweeteners development. In this study, a xerophilic fungus Aspergillus niger GH1 isolated of the Mexican semi-desert, previously reported as an invertase over-producer strain was used. Molasses from Mexico and Cuba were chemically analyzed (total and reducer sugars, nitrogen and phosphorous contents); the last one was selected based on chemical composition. Fermentations were performed using virgin and hydrolyzate bagasse (treatment with concentrated sulfuric acid). Results indicated that, the enzymatic yield (5231 U/L) is higher than those reported by other A. niger strains under solid state fermentation, using hydrolyzate bagasse. The acid hydrolysis promotes availability of fermentable sugars. In addition, maximum invertase activity was detected at 24 h using low substrate concentration, which may reduce production costs. This study presents an alternative method for invertase production using a xerophilic fungus isolated from Mexican semi-desert and inexpensive substrates (molasses and sugarcane bagasse). PMID:25242918

Veana, F.; Martínez-Hernández, J.L.; Aguilar, C.N.; Rodríguez-Herrera, R.; Michelena, G.

2014-01-01

226

The use of RFLP analysis in classification of the black Aspergilli: reinterpretation of the Aspergillus niger aggregate  

Microsoft Academic Search

By studying ribosomal banding patterns in ethidium bromide-stained gels of chromosomal digests we are able to provide a rapid and reliable classification of a number of taxonomically important isolates of the black Aspergilli. This classification is supported by Southern blots using several pectin lyase genes, isolated from A. niger CBS 120.49, as probes. Taxonomy on the basis of RFLP analysis

Margo A. Kusters-van Someren; Robert A. Samson; Jaap Visser

1991-01-01

227

Mixed Disulfide Formation at Cys141 Leads to Apparent Unidirectional Attenuation of Aspergillus niger NADP-Glutamate Dehydrogenase Activity  

PubMed Central

NADP-Glutamate dehydrogenase from Aspergillus niger (AnGDH) exhibits sigmoid 2-oxoglutarate saturation. Incubation with 2-hydroxyethyl disulfide (2-HED, the disulfide of 2-mercaptoethanol) resulted in preferential attenuation of AnGDH reductive amination (forward) activity but with a negligible effect on oxidative deamination (reverse) activity, when monitored in the described standard assay. Such a disulfide modified AnGDH displaying less than 1.0% forward reaction rate could be isolated after 2-HED treatment. This unique forward inhibited GDH form (FIGDH), resembling a hypothetical ‘one-way’ active enzyme, was characterized. Kinetics of 2-HED mediated inhibition and protein thiol titrations suggested that a single thiol group is modified in FIGDH. Two site-directed cysteine mutants, C141S and C415S, were constructed to identify the relevant thiol in FIGDH. The forward activity of C141S alone was insensitive to 2-HED, implicating Cys141 in FIGDH formation. It was observed that FIGDH displayed maximal reaction rate only after a pre-incubation with 2-oxoglutarate and NADPH. In addition, compared to the native enzyme, FIGDH showed a four fold increase in K0.5 for 2-oxoglutarate and a two fold increase in the Michaelis constants for ammonium and NADPH. With no change in the GDH reaction equilibrium constant, the FIGDH catalyzed rate of approach to equilibrium from reductive amination side was sluggish. Altered kinetic properties of FIGDH at least partly account for the observed apparent loss of forward activity when monitored under defined assay conditions. In sum, although Cys141 is catalytically not essential, its covalent modification provides a striking example of converting the biosynthetic AnGDH into a catabolic enzyme. PMID:24987966

Walvekar, Adhish S.; Choudhury, Rajarshi; Punekar, Narayan S.

2014-01-01

228

Replacement P212H Altered the pH-Temperature Profile of Phytase from Aspergillus niger NII 08121.  

PubMed

Microbial phytase, a widely used animal feed enzyme, needs to be active and stable in the acidic milieu for better performance in the monogastric gut. Aspergillus niger phytases exhibit an activity dip in the pH range from 3.0 to 3.5. Replacement of amino acids, which changed the pKa of catalytic residues H82 and D362, resulted in alteration of the pH profile of a thermostable phytase from A. niger NII 08121. Substitution P212H in the protein loop at 14 Å distance to the active site amended the pH optimum from 2.5 to pH 3.2 nevertheless with a decrease in thermostability than the wild enzyme. This study described the utility of amino acid replacements based on pKa shifts of catalytic acid/base to modulate the pH profile of phytases. PMID:25595493

Ushasree, Mrudula Vasudevan; Vidya, Jalaja; Pandey, Ashok

2015-03-01

229

Bioconversion of waste office paper to gluconic acid in a turbine blade reactor by the filamentous fungus Aspergillus niger.  

PubMed

Gluconic acid production was investigated using an enzymatic hydrolysate of waste office automation paper in a culture of Aspergillus niger. In repeated batch cultures using flasks, saccharified solution medium (SM) did not show any inhibitory effects on gluconic acid production compared to glucose medium (GM). The average gluconic acid yields were 92% (SM) and 80% (GM). In repeated batch cultures using SM in a turbine blade reactor (TBR), the gluconic acid yields were 60% (SM) and 67% (GM) with 80-100 g/l of gluconic acid. When pure oxygen was supplied the production rate increased to four times higher than when supplying air. Remarkable differences in the morphology of A. niger and dry cell weight between SM and GM were observed. The difference in morphology may have caused a reduction of oxygen transfer, resulting in a decrease in gluconic acid production rate in SM. PMID:15979872

Ikeda, Yuko; Park, Enock Y; Okuda, Naoyuki

2006-05-01

230

Production of the Phanerochaete flavido-alba laccase in Aspergillus niger for synthetic dyes decolorization and biotransformation.  

PubMed

We investigated the expression of Phanerochaete flavido-alba laccase gene in Aspergillus niger and the physical and biochemical properties of the recombinant enzyme (rLac-LPFA) in order to test it for synthetic dye biotransformation. A. niger was able to produce high levels of active recombinant enzyme (30 mgL(-1)), whose identity was further confirmed by immunodetection using Western blot analysis and N-terminal sequencing. Interestingly, rLac-LPFA exhibited an improved stability at pH (2-9) and organic solvents tested. Furthermore, the percentage of decoloration and biotransformation of synthetic textile dyes, Remazol Brilliant Blue R (RBBR) and Acid Red 299 (NY1), was higher than for the native enzyme. Its high production, simple purification, high activity, stability and ability to transform textile dyes make rLac-LPFA a good candidate for industrial applications. PMID:23884844

Benghazi, Lamiae; Record, Eric; Suárez, Antonio; Gomez-Vidal, José A; Martínez, José; de la Rubia, Teresa

2014-01-01

231

Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition*  

PubMed Central

Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization. Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each significant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by multiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x 1 (urea)=0.163 (41.63 g/L), x 2 (Na2CO3)=?1.68 (2.64 g/L), x 3 (MgSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization. PMID:18600786

Xu, Yao-xing; Li, Yan-li; Xu, Shao-chun; Liu, Yong; Wang, Xin; Tang, Jiang-wu

2008-01-01

232

Physical and Kinetic Properties of the Family 3 ?-Glucosidase from Aspergillus niger Which Is Important for Cellulose Breakdown  

Microsoft Academic Search

A ß-glucosidase (BGS) purified from Aspergillus niger cellulase powder (obtained from Sigma, St. Louis, MO, USA) was characterized. Electrophoresis, size exclusion chromatography, and dynamic light scattering indicated that the enzyme is a dimer of approximately 200 kDa. Five of the seven N-glycosylated oligosaccharides attached to BGS were composed of D-mannoses attached to a ß(1-4)-N-acetyl-glucosamine-ß-(1-4)-fucose-a-(1-6)-N-acetylglucosamine core. The other two were similar,

Heather F. Seidle; Ira Marten; Oded Shoseyov; Reuben E. Huber

2004-01-01

233

Thermal stability of Trichoderma reesei c30 cellulase and aspergillus niger; -glucosidase after ph and chemical modification  

SciTech Connect

Treatment of Trichoderma reesei C30 cellulase at pH 10.0 for 1 h at room temperature increased its pH and thermal stability. Chemical modification of the free epsilon-amino groups of cellulase at pH 10.0 resulted in no further increase in stability. Such chemical modification, however, decreased the thermal stability of the cellulose-cellulase complex. On the contrary, the chemical modification of Aspergillus niger glucosidase with glutaraldehyde at pH 8.0 increased the thermal stability of this enzyme.

Woodward, J.; Whaley, K.S.; Zachry, G.S.; Wohlpart, D.L.

1981-01-01

234

Crystal structure of Aspergillus niger isopullulanase, a member of glycoside hydrolase family 49.  

PubMed

An isopullulanase (IPU) from Aspergillus niger ATCC9642 hydrolyzes alpha-1,4-glucosidic linkages of pullulan to produce isopanose. Although IPU does not hydrolyze dextran, it is classified into glycoside hydrolase family 49 (GH49), major members of which are dextran-hydrolyzing enzymes. IPU is highly glycosylated, making it difficult to obtain its crystal. We used endoglycosidase H(f) to cleave the N-linked oligosaccharides of IPU, and we here determined the unliganded and isopanose-complexed forms of IPU, both solved at 1.7-A resolution. IPU is composed of domains N and C joined by a short linker, with electron density maps for 11 or 12 N-acetylglucosamine residues per molecule. Domain N consists of 13 beta-strands and forms a beta-sandwich. Domain C, where the active site is located, forms a right-handed beta-helix, and the lengths of the pitches of each coil of the beta-helix are similar to those of GH49 dextranase and GH28 polygalacturonase. The entire structure of IPU resembles that of a GH49 enzyme, Penicillium minioluteum dextranase (Dex49A), despite a difference in substrate specificity. Compared with the active sites of IPU and Dex49A, the amino acid residues participating in subsites +2 and +3 are not conserved, and the glucose residues of isopanose bound to IPU completely differ in orientation from the corresponding glucose residues of isomaltose bound to Dex49A. The shape of the catalytic cleft characterized by the seventh coil of the beta-helix and a loop from domain N appears to be critical in determining the specificity of IPU for pullulan. PMID:18155243

Mizuno, Masahiro; Koide, Atsushi; Yamamura, Akihiro; Akeboshi, Hiromi; Yoshida, Hiromi; Kamitori, Shigehiro; Sakano, Yoshiyuki; Nishikawa, Atsushi; Tonozuka, Takashi

2008-02-01

235

Purification and characterization of endo-xylanases from aspergillus Niger. II. An enzyme of PL 45  

SciTech Connect

A homogeneous endo-xylanase (1,4-..beta..-D-xylan xylano-hydrolase, EC 3.2.1.8) was obtained from a crude Aspergillus niger pentosanase by chromatography with Ultrogel AcA 54, SP-Sephadex C-25 at pH 4.5, DEAE-Sephadex A-25 at pH 5.4, Sephadex G-50, and SP-Sephadex C-25 with a gradient from pH 2.8 to pH 4.6. It was much more active on soluble than on insoluble xylan yielding large amounts of unreacted xylan and a mixture of oligosaccharides with chain lengths from two to six. No xylose or L-arabinose was produced. There was high activity on a xylopentaose through xylononaose mixture, but not on xylobiose, xylotriose, or xylotetraose. The enzyme had slight activity on untreated cellulose, carboxymethylcellulose, and pectin. Molecular weight was ca. 1.4 x 10/sup 4/, with an isoelectric point of 4.5 and an amino acid profile high in acidic but low in sulfur-containing residues. In a 25-min assay at pH 4.7, this endo-xylanase was most active at 45 degrees C, with an activation energy from 5 to 35 degrees C of 33.3 kJ/mol. The optimum pH for activity was 4.9. Decay in buffer was first order, with an activation energy at pH 4.7 from 48 to 53 degrees C of 460 kJ/mol. Optimum pH for stability was about 5.6, where the half-life at 48 degrees C in buffer was ca. 40 h.

Shei, J.C.; Fratzke, A.R.; Frederick, M.M.; Frederick, J.R.; Reilly, P.J.

1985-04-01

236

Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.  

PubMed

The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 ?g/g and 7.0 ?g/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes. PMID:25364929

Passamani, Fabiana Reinis Franca; Hernandes, Thais; Lopes, Noelly Alves; Bastos, Sabrina Carvalho; Santiago, Wilder Douglas; Cardoso, Maria das Graças; Batista, Luís Roberto

2014-11-01

237

Physiochemical properties and kinetics of glucoamylase produced from deoxy-d-glucose resistant mutant of Aspergillus niger for soluble starch hydrolysis  

PubMed Central

Glucoamylases (GAs) from a wild and a deoxy-d-glucose-resistant mutant of a locally isolated Aspergillus niger were purified to apparent homogeneity. The subunit molecular mass estimated by SDS–PAGE was 93 kDa for both strains, while the molecular masses determined by MALDI-TOF for wild and mutant GAs were 72.876 and 72.063 kDa, respectively. The monomeric nature of the enzymes was confirmed through activity staining. Significant improvement was observed in the kinetic properties of the mutant GA relative to the wild type enzyme. Kinetic constants of starch hydrolysis for A. niger parent and mutant GAs calculated on the basis of molecular masses determined through MALDI-TOF were as follows: kcat = 343 and 727 s?1, Km = 0.25 and 0.16 mg mL?1, kcat/Km (specificity constant) = 1374 and 4510 mg mL?1 s?1, respectively. Thermodynamic parameters for soluble starch hydrolysis also suggested that mutant GA was more efficient compared to the parent enzyme. PMID:24293795

Riaz, Muhammad; Rashid, Muhammad Hamid; Sawyer, Lindsay; Akhtar, Saeed; Javed, Muhammad Rizwan; Nadeem, Habibullah; Wear, Martin

2012-01-01

238

Physiochemical properties and kinetics of glucoamylase produced from deoxy-d-glucose resistant mutant of Aspergillus niger for soluble starch hydrolysis.  

PubMed

Glucoamylases (GAs) from a wild and a deoxy-d-glucose-resistant mutant of a locally isolated Aspergillus niger were purified to apparent homogeneity. The subunit molecular mass estimated by SDS-PAGE was 93 kDa for both strains, while the molecular masses determined by MALDI-TOF for wild and mutant GAs were 72.876 and 72.063 kDa, respectively. The monomeric nature of the enzymes was confirmed through activity staining. Significant improvement was observed in the kinetic properties of the mutant GA relative to the wild type enzyme. Kinetic constants of starch hydrolysis for A. niger parent and mutant GAs calculated on the basis of molecular masses determined through MALDI-TOF were as follows: k cat = 343 and 727 s(-1), K m = 0.25 and 0.16 mg mL(-1), k cat/K m (specificity constant) = 1374 and 4510 mg mL(-1) s(-1), respectively. Thermodynamic parameters for soluble starch hydrolysis also suggested that mutant GA was more efficient compared to the parent enzyme. PMID:24293795

Riaz, Muhammad; Rashid, Muhammad Hamid; Sawyer, Lindsay; Akhtar, Saeed; Javed, Muhammad Rizwan; Nadeem, Habibullah; Wear, Martin

2012-01-01

239

Identification and susceptibility of Aspergillus section nigri in china: prevalence of species and paradoxical growth in response to echinocandins.  

PubMed

Molecular identification and in vitro antifungal susceptibility tests of 43 Aspergillus section Nigri isolates from China were performed. Aspergillus niger and Aspergillus tubingensis were present in almost equal numbers. All of the isolates had low MIC/MECs (minimum effective concentrations) for the 7 common antifungals, and a paradoxical effect was observed for the first time in response to caspofungin and micafungin. PMID:25502526

Li, Yali; Wan, Zhe; Liu, Wei; Li, Ruoyu

2015-02-01

240

Aspergillus pragensis sp. nov. discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi.  

PubMed

The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of ?-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspected and proven onychomycosis, one from otitis externa, and two associated with probable invasive aspergillosis. The results showed that one Aspergillus candidus isolate was the cause of otitis externa, and both isolates obtained from sputa of patients with probable invasive aspergillosis were reidentified as A. carneus (sect. Terrei) and A. flavus (sect. Flavi). Three isolates from nail scrapings were identified as A. tritici, a verified agent of nondermatophyte onychomycosis. One isolate from toenail was determined to be A. candidus and the two isolates belonged to a hitherto undescribed species, Aspergillus pragensis sp. nov. This species is well supported by phylogenetic analysis based on ?-tubulin and calmodulin gene and is distinguishable from other members of sect. Candidi by red-brown reverse on malt extract agar, slow growth on Czapek-Dox agar and inability to grow at 37°C. A secondary metabolite analysis was also provided with comparison of metabolite spectrum to other species. Section Candidi now encompasses five species for which a dichotomous key based on colony characteristics is provided. All clinical isolates were tested for susceptibilities to selected antifungal agents using the Etest and disc diffusion method. Overall sect. Candidi members are highly susceptible to common antifungals. PMID:24951723

Hubka, Vit; Lyskova, Pavlina; Frisvad, Jens C; Peterson, Stephen W; Skorepova, Magdalena; Kolarik, Miroslav

2014-08-01

241

Atomistic details of effect of disulfide bond reduction on active site of Phytase B from Aspergillus niger: A MD Study  

PubMed Central

The molecular integrity of the active site of phytases from fungi is critical for maintaining phytase function as efficient catalytic machines. In this study, the molecular dynamics (MD) of two monomers of phytase B from Aspergillus niger, the disulfide intact monomer (NAP) and a monomer with broken disulfide bonds (RAP), were simulated to explore the conformational basis of the loss of catalytic activity when disulfide bonds are broken. The simulations indicated that the overall secondary and tertiary structures of the two monomers were nearly identical but differed in some crucial secondary–structural elements in the vicinity of the disulfide bonds and catalytic site. Disulfide bonds stabilize the ?-sheet that contains residue Arg66 of the active site and destabilize the ?-helix that contains the catalytic residue Asp319. This stabilization and destabilization lead to changes in the shape of the active–site pocket. Functionally important hydrogen bonds and atomic fluctuations in the catalytic pocket change during the RAP simulation. None of the disulfide bonds are in or near the catalytic pocket but are most likely essential for maintaining the native conformation of the catalytic site. Abbreviations PhyB - 2.5 pH acid phophatese from Aspergillus niger, NAP - disulphide intact monomer of Phytase B, RAP - disulphide reduced monomer of Phytase B, Rg - radius of gyration, RMSD - root mean square deviation, MD - molecular dynamics. PMID:24391358

Kumar, Kapil; Dixit, Mudit; Khire, JM; Pal, Sourav

2013-01-01

242

Genome-wide transcriptional response of Trichoderma reesei to lignocellulose using RNA sequencing and comparison with Aspergillus niger  

PubMed Central

Background A major part of second generation biofuel production is the enzymatic saccharification of lignocellulosic biomass into fermentable sugars. Many fungi produce enzymes that can saccarify lignocellulose and cocktails from several fungi, including well-studied species such as Trichoderma reesei and Aspergillus niger, are available commercially for this process. Such commercially-available enzyme cocktails are not necessarily representative of the array of enzymes used by the fungi themselves when faced with a complex lignocellulosic material. The global induction of genes in response to exposure of T. reesei to wheat straw was explored using RNA-seq and compared to published RNA-seq data and model of how A. niger senses and responds to wheat straw. Results In T. reesei, levels of transcript that encode known and predicted cell-wall degrading enzymes were very high after 24 h exposure to straw (approximately 13% of the total mRNA) but were less than recorded in A. niger (approximately 19% of the total mRNA). Closer analysis revealed that enzymes from the same glycoside hydrolase families but different carbohydrate esterase and polysaccharide lyase families were up-regulated in both organisms. Accessory proteins which have been hypothesised to possibly have a role in enhancing carbohydrate deconstruction in A. niger were also uncovered in T. reesei and categories of enzymes induced were in general similar to those in A. niger. Similarly to A. niger, antisense transcripts are present in T. reesei and their expression is regulated by the growth condition. Conclusions T. reesei uses a similar array of enzymes, for the deconstruction of a solid lignocellulosic substrate, to A. niger. This suggests a conserved strategy towards lignocellulose degradation in both saprobic fungi. This study provides a basis for further analysis and characterisation of genes shown to be highly induced in the presence of a lignocellulosic substrate. The data will help to elucidate the mechanism of solid substrate recognition and subsequent degradation by T. reesei and provide information which could prove useful for efficient production of second generation biofuels. PMID:24060058

2013-01-01

243

Enzymatic synthesis of fructooligosaccharides by inulinases from Aspergillus niger and Kluyveromyces marxianus NRRL Y-7571 in aqueous-organic medium.  

PubMed

This work is focused on the synthesis of the fructooligosaccharides (FOS) from sucrose and inulin, using free, immobilized and pre-treated immobilized inulinase from Kluyveromyces marxianus NRRL Y 7571 and Aspergillus niger in an aqueous-organic system. Initially, the influence of pre-treatment using four different gases, propane, n-butane, CO(2) and liquefied petroleum gas (LPG), was investigated towards FOS production and best results were found when both enzymes were previously treated with LPG. The best reaction yields were obtained when the immobilized enzymes were treated with LPG. Considering FOS synthesis using the enzyme from A. niger, yields of 26.62% of GF2 (kestose), 30.62% of GF3 (nystose) and 8.47% of GF4 (fructosyl nystose) were achieved using sucrose as substrate. Using inulinases from K. marxianus NRRL Y 7571, 11.89% of GF2 and 20.83% of GF3 were obtained, using inulin as substrate. However, promising results were achieved using the free form of inulinase from A. niger (77.19% of GF2; 14.03% of GF3 and 0.07% of GF4) using inulin as substrate. PMID:23265469

Silva, Marceli Fernandes; Rigo, Diane; Mossi, Vinícius; Golunski, Simone; Kuhn, Graciele de Oliveira; Di Luccio, Marco; Dallago, Rogério; de Oliveira, Débora; Oliveira, J Vladimir; Treichel, Helen

2013-05-01

244

The effect of various food parameters on the activity and stability of catalase from Aspergillus niger and catalase from bovine liver  

Microsoft Academic Search

The effects of a number of food relevant parameters on catalase activity and stability were studied. The direct responses of different combinations of the parameters ethanol, pH and ionic strength on bovine liver catalase and Aspergillus niger catalase activity were investigated in a full factorial 24 statistically designed experiment. Statistically significant effects (p = 0.001) on both types of catalases

Anne S. Meyer; Lærke H. Pedersen; Anette Isaksen

1997-01-01

245

Establishment of Two Ectomycorrhizal Shrub Species in a Semiarid Site after in Situ Amendment with Sugar Beet, Rock Phosphate, and Aspergillus niger  

Microsoft Academic Search

A field experiment was carried out to assess the effectiveness of the addition of sugar beet, rock phosphate, and Aspergillus niger directly into the planting hole, and the mycorrhizal inoculation of seedlings with Scleroderma verrucosum, for promotion of plant growth of Cistus albidus L. and Quercus coccifera L. and enhancement of soil physicochemical, biochemical, and biological properties, in a degraded

F. Caravaca; M. M. Alguacil; R. Azcón; J. Parladé; P. Torres; A. Roldán

2005-01-01

246

Correlation of Mycotoxin Fumonisin B2 Production and Presence of the Fumonisin Biosynthetic Gene fum8 in Aspergillus niger from Grape  

Technology Transfer Automated Retrieval System (TEKTRAN)

Fumonisins are mycotoxins associated with cancer and several other serious diseases in humans and animals. Production of the mycotoxins has been reported for over two decades in Fusarium species, but has been reported only recently in strains of Aspergillus niger. In addition, a homologue of the f...

247

SHIFTING THE PH PROFILE OF ASPERGILLUS NIGER PHYA PHYTASE TO MATCH THE STOMACH PH ENHANCES ITS EFFECTIVENESS AS AN ANIMAL FEED ADDITIVE  

Technology Transfer Automated Retrieval System (TEKTRAN)

Environmental pollution of phosphorus (P) from animal waste is a major problem in agriculture because simple-stomached animals such as swine, poultry, and fish cannot digest phosphorus (as phytate) present in plant feeds. To alleviate this problem, a phytase from Aspergillus niger PhyA is widely us...

248

Enhanced enzyme production from mixed cultures of Trichoderma reesei RUT-C30 and Aspergillus niger LMA grown as fed batch in a stirred tank bioreactor  

Microsoft Academic Search

For the complete hydrolysis of cellulose, the cellulolytic fungi produce a whole set of commercially important enzymes called cellulases. The aim of this work was to investigate an approach to enhance the production of these enzymes by co-culturing Trichoderma reesei and Aspergillus niger in a bioreactor to convert cellulose substrate into soluble sugars through a synergetic action of enzyme complex

Aftab Ahamed; Patrick Vermette

2008-01-01

249

Altering the Substrate Specificity Site of Aspergillus Niger PhyB shifts the pH optimum to pH 3.2  

Technology Transfer Automated Retrieval System (TEKTRAN)

Phytases are of biotechnological importance as animal feed additives for their ability to catalyze the hydrolysis of phosphate from phytate for absorption by simple-stomached animals, and to reduce their fecal phosphorus excretion. Aspergillus niger PhyB has high catalytic activity at low pHs around...

250

Characterization of a polyketide synthase in Aspergillus niger whose product is a precursor for both dihydroxynaphthalene (DHN) melanin and naphtho-?-pyrone  

PubMed Central

The genome sequencing of the fungus Aspergillus niger uncovered a large cache of genes encoding enzymes thought to be involved in the production of secondary metabolites yet to be identified. Identification and structural characterization of many of these predicted secondary metabolites are hampered by their low concentration relative to the known A. niger metabolites such as the naphtho-?-pyrone family of polyketides. We deleted a nonreducing PKS gene in A. niger strain ATCC 11414, a daughter strain of A. niger ATCC strain 1015 whose genome was sequenced by the DOE Joint Genome Institute. This PKS encoding gene we name albA is a predicted ortholog of alb1 from Aspergillus fumigatus which is responsible for production of the naphtho-?-pyrone precursor for the 1,8-dihydroxynaphthalene (DHN) melanin/spore pigment. Our results show that the A. nigeralbA PKS is responsible for both the production of the spore pigment precursor and a family of naphtho-?-pyrones commonly found in significant quantity in A. niger culture extracts. The generation of an A. niger strain devoid of naphtho-?-pyrones will greatly facilitate the elucidation of cryptic biosynthetic pathways in this organism. PMID:21176790

Chiang, Yi-Ming; Meyer, Kristen M.; Praseuth, Michael; Baker, Scott E.; Bruno, Kenneth S.; Wang, Clay C. C.

2011-01-01

251

Characterization of a polyketide synthase in Aspergillus niger whose product is a precursor for both dihydroxynaphthalene (DHN) melanin and naphtho-?-pyrone.  

PubMed

The genome sequencing of the fungus Aspergillus niger uncovered a large cache of genes encoding enzymes thought to be involved in the production of secondary metabolites yet to be identified. Identification and structural characterization of many of these predicted secondary metabolites are hampered by their low concentration relative to the known A. niger metabolites such as the naphtho-?-pyrone family of polyketides. We deleted a non-reducing PKS gene in A. niger strain ATCC 11414, a daughter strain of A. niger ATCC strain 1015 whose genome was sequenced by the DOE Joint Genome Institute. This PKS encoding gene we name albA is a predicted ortholog of alb1 from Aspergillus fumigatus which is responsible for production of the naphtho-?-pyrone precursor for the 1,8-dihydroxynaphthalene (DHN) melanin/spore pigment. Our results show that the A. nigeralbA PKS is responsible for both the production of the spore pigment precursor and a family of naphtho-?-pyrones commonly found in significant quantity in A. niger culture extracts. The generation of an A. niger strain devoid of naphtho-?-pyrones will greatly facilitate the elucidation of cryptic biosynthetic pathways in this organism. PMID:21176790

Chiang, Yi-Ming; Meyer, Kristen M; Praseuth, Michael; Baker, Scott E; Bruno, Kenneth S; Wang, Clay C C

2011-04-01

252

Characterization of a polyketide synthase in Aspergillus niger whose product is a precursor for both dihydroxynaphthalene (DHN) melanin and naphtho-?-pyrone.  

SciTech Connect

The genome sequencing of the fungus Aspergillus niger, an industrial workhorse, uncovered a large cache of genes encoding enzymes thought to be involved in the production of secondary metabolites yet to be identified. Identification and structural characterization of many of these predicted secondary metabolites are hampered by their low concentration relative to the known A. niger metabolites such as the naphtho-?-pyrone family of polyketides. We deleted a nonreducing PKS gene in A. niger strain ATCC 11414, a daughter strain of A. niger ATCC strain 1015 whose genome was sequenced by the DOE Joint Genome Institute. This PKS encoding gene is a predicted ortholog of alb1 from Aspergillus fumigatus which is responsible for production of YWA1, a precursor of fungal DHN melanin. Our results show that the A. niger alb1 PKS is responsible for the production of the polyketide precursor for DHN melanin biosynthesis. Deletion of alb1 elimnates the production of major metabolites, naphtho-?-pyrones. The generation of an A. niger strain devoid of naphtho-?-pyrones will greatly facilitate the elucidation of cryptic biosynthetic pathways in this organism.

Chiang, Yi Ming; Meyer, Kristen M.; Praseuth , Michael; Baker, Scott E.; Bruno, Kenneth S.; Wang, Clay C.

2010-12-06

253

Comparative study of toxicity of azo dye Procion Red MX-5B following biosorption and biodegradation treatments with the fungi Aspergillus niger and Aspergillus terreus.  

PubMed

Azo dyes are an important class of environmental contaminants and are characterized by the presence of one or more azo bonds (-N=N-) in their molecular structure. Effluents containing these compounds resist many types of treatments due to their molecular complexity. Therefore, alternative treatments, such as biosorption and biodegradation, have been widely studied to solve the problems caused by these substances, such as their harmful effects on the environment and organisms. The aim of the present study was to evaluate biosorption and biodegradation of the azo dye Procion Red MX-5B in solutions with the filamentous fungi Aspergillus niger and Aspergillus terreus. Decolorization tests were performed, followed by acute toxicity tests using Lactuca sativa seeds and Artemia salina larvae. Thirty percent dye removal of the solutions was achieved after 3 h of biosorption. UV-Vis spectroscopy revealed that removal of the dye molecules occurred without major molecular changes. The acute toxicity tests confirmed lack of molecular degradation following biosorption with A. niger, as toxicity to L. sativa seed reduced from 5% to 0%. For A. salina larvae, the solutions were nontoxic before and after treatment. In the biodegradation study with the fungus A. terreus, UV-Vis and FTIR spectroscopy revealed molecular degradation and the formation of secondary metabolites, such as primary and secondary amines. The biodegradation of the dye molecules was evaluated after 24, 240 and 336 h of treatment. The fungal biomass demonstrated considerable affinity for Procion Red MX-5B, achieving approximately 100% decolorization of the solutions by the end of treatment. However, the solutions resulting from this treatment exhibited a significant increase in toxicity, inhibiting the growth of L. sativa seeds by 43% and leading to a 100% mortality rate among the A. salina larvae. Based on the present findings, biodegradation was effective in the decolorization of the samples, but generated toxic metabolites, while biosorption was effective in both decolorization and reducing the toxicity of the solutions. PMID:25048922

Almeida, E J R; Corso, C R

2014-10-01

254

Isolation and characterization of ochratoxin A and aflatoxin B1 producing fungi infecting grapevines cultivated in Tunisia  

Microsoft Academic Search

A survey was conducted to access mycotoxin-producing fungi and to evaluate the ochratoxin A (OTA) and the Aflatoxin B1 (AFB1) potential production for fungal strains contaminating table grapes in different Tunisian vineyards. Among 100 Aspergillus isolates, Aspergillus niger aggregate were the most frequent (70%) followed by Aspergillus carbonarius (7%) and Aspergillus flavus (23%). The mycotoxigenic capacity of the isolates was

S. Melki; Ben Fredj; S. Chebil; A. Mliki

2009-01-01

255

Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels  

PubMed Central

Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE), in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (?-glucosidase, endoglucanase and filterpaperase) using a novel substrate, Banana Peels (BP) for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes. PMID:25763058

Rehman, Shazia; Aslam, Hina; Ahmad, Aqeel; Khan, Shakeel Ahmed; Sohail, Muhammad

2014-01-01

256

Phase diagram of crystallization of Aspergillus niger acid proteinase A, a non-pepsin-type acid proteinase  

NASA Astrophysics Data System (ADS)

Proteinase A from Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase with an extremely low isoelectric point (pI 3.3). The protein is crystallized from ammonium sulfate solutions of pH lower than 4. The crystallization is affected by the presence of dimethylsulfoxide (DMSO). We have studied the phase diagram of the crystallization of proteinase A in the absence and presence of DMSO, to clarify crystallization at such an extremely low pH and to study the effects of DMSO. The results indicate that the logarithm of protein solubility is a rectilinear function of ammonium sulfate concentration in both the absence and presence of DMSO. DMSO definitely lowers the solubility at relatively low concentrations of ammonium sulfate, but had little effect on protein solubility at higher concentrations of ammonium sulfate.

Kudo, Norio; Ataka, Mitsuo; Sasaki, Hiroshi; Muramatsu, Tomonari; Katsura, Tatsuo; Tanokura, Masaru

1996-10-01

257

A qualitative and quantitative high-throughput assay for screening of gluconate high-yield strains by Aspergillus niger.  

PubMed

A novel two-step high-throughput strategy was developed for screening of gluconate high-yield strains by Aspergillus niger. The first step was fast qualitative assay according to the indicator color change, the second step was quantitative assay according to the absorbance of chelate formed with Cu(2+) at 810nm. The accuracy of high-throughput assay was comparable to that of high-performance liquid chromatography (HPLC). The correlation coefficient between CuSO4 assay and HPLC assays was exceeding 0.99 by statistical analysis. As a result, 3 high-yield mutants were screened out from 1000 viable single colonies, the mutants II-2-A1, IV-7-C6, and V-11-C5 were further validated in 5L of bioreactor. The average production rates were 15.5%, 32.8%, and 12.1% higher than that of the parental strain, respectively. PMID:25498457

Shi, Fei; Tan, Jun; Chu, Ju; Wang, Yonghong; Zhuang, Yingping; Zhang, Siliang

2015-02-01

258

Gamma radiation induced mutagenesis in Aspergillus niger to enhance its microbial fermentation activity for industrial enzyme production.  

PubMed

?- and ?-Galactosidases find application in food processing, health and nutrition. Aspergillus niger is one of the potent producer of these enzymes and was genotypically improved using gamma-ray induced mutagenesis. The mutant-derivative produced two-fold higher ?- and ?-galactosidases. For testing genetic variability and its relationship with phenotypic properties of the two organisms, DNA samples of the mutant and parental strains of A. niger were amplified with 28 deca-nucleotide synthetic primers. RAPD analysis showed significantly different pattern between parental and mutant cultures. The mutant derivative yielded homogeneous while parental strain formed heterogeneous amplification patterns. Seven primers identified 42.9% polymorphism in the amplification products, indicating that these primers determined some genetic variability between the two strains. Thus RAPD was found to be an efficient technique to determine genetic variability in the mutant and wild organisms. Both wild and mutant strains were analyzed for their potential to produce galactosidases. Comparison of different carbon sources on enzyme yield revealed that wheat bran is significant (P < 0.01) effective producer and economical source followed by rice bran, rice polishing and lactose. The mutant was significantly better enzyme producer and could be considered for its prospective application in food, nutrition and health and that RAPD can be effectively used to differentiate mutant strain from the parental strain based on the RAPD patterns. PMID:20632114

Awan, M Siddique; Tabbasam, Nabila; Ayub, N; Babar, M E; Mehboob-ur-Rahman; Rana, Shahid Mahboob; Rajoka, M I

2011-02-01

259

An acidothermophilic functionally active novel GH12 family endoglucanase from Aspergillus niger HO: purification, characterization and molecular interaction studies.  

PubMed

Endoglucanase (EG) from Aspergillus niger HO was sequentially purified through ultrafiltration, ion exchange and size exclusion chromatography to homogeneity, with an overall recovery of 18 %. The purified EG was a monomeric protein with a molecular weight of about 55 kDa. The enzyme was optimally active at pH 3.5 and 70 °C with a half life (t1/2) of 3 h and Km value of 2.5 mg/ml. Metal ions, such as Ca(2+) and Co(2+) helped in enzyme induction, while Hg(2+) and Cu(2+) strongly inhibited the enzyme activity. Peptide mass fingerprinting results revealed that the purified EG is a novel enzyme that belongs to family 12 of glycoside hydrolase (GH12). Molecular docking studies indicated the presence of Glu116 and Glu204 as important determinant residues for the functional interaction with carboxymethylcellulose and showed hydrogen bonding with Asp99, Glu116, Glu204 and hydrophobic interactions with Trp22, Val58, Tyr61, Phe101, Met118, Trp120, Pro129, Ile130, Thr160 and Phe206. Hydrolysis of 2 % CMC with purified acidothermophilic EG at its optimum temperature and pH resulted in complete hydrolysis within 2 h yielding 18 % cellotriose, 72 % cellobiose and 10 % glucose as evident from HPLC analysis. In comparison to most of the EGs reported in literature, EG from A. niger HO exhibited higher thermostability. The acidothermophilic nature of this enzyme makes it potentially useful for industrial applications. PMID:25331339

Rawat, Rekha; Kumar, Sunil; Chadha, Bhupinder Singh; Kumar, Dinesh; Oberoi, Harinder Singh

2015-01-01

260

Mechanism of Cr(VI) reduction by Aspergillus niger: enzymatic characteristic, oxidative stress response, and reduction product.  

PubMed

Bioremediation of hexavalent chromium by Aspergillus niger was attributed to the reduction product (trivalent chromium) that could be removed in precipitation and immobilized inside the fungal cells and on the surface of mycelium. The site location of reduction was conducted with assays of the permeabilized cells, cell-free extracts, and cell debris, which confirmed that the chromate reductase was mainly located in the soluble fraction of cells. The oxidation-reduction process was accompanied by the increase of reactive oxygen species and antioxidant levels after hexavalent chromium treatment. Michaelis-Menten constant (K m) and maximum reaction rate (V max), obtained from the Lineweaver-Burk plot were 14.68 ?M and 434 ?M min(-1) mg(-1) of protein, respectively. Scanning electron microscopy and Raman spectra analyses manifested that both Cr(VI) and Cr(III) species were present on the mycelium. Fourier transform-infrared spectroscopy analysis suggested that carboxyl, hydroxide, amine, amide, cyano-group, and phosphate groups from the fungal cell wall were involved in chromium binding by the complexation with the Cr(III) and Cr(VI) species. A Cr(VI) removal mechanism of Cr(VI) reduction followed by the surface immobilization and intracellular accumulation of Cr(III) in living A. niger was present. PMID:25408081

Gu, Yanling; Xu, Weihua; Liu, Yunguo; Zeng, Guangming; Huang, Jinhui; Tan, Xiaofei; Jian, Hao; Hu, Xi; Li, Fei; Wang, Dafei

2015-04-01

261

Conversion of orange peel to L-galactonic acid in a consolidated process using engineered strains of Aspergillus niger  

PubMed Central

Citrus processing waste is a leftover from the citrus processing industry and is available in large amounts. Typically, this waste is dried to produce animal feed, but sometimes it is just dumped. Its main component is the peel, which consists mostly of pectin, with D-galacturonic acid as the main monomer. Aspergillus niger is a filamentous fungus that efficiently produces pectinases for the hydrolysis of pectin and uses the resulting D-galacturonic acid and most of the other components of citrus peel for growth. We used engineered A. niger strains that were not able to catabolise D-galacturonic acid, but instead converted it to L-galactonic acid. These strains also produced pectinases for the hydrolysis of pectin and were used for the conversion of pectin in orange peel to L-galactonic acid in a consolidated process. The D-galacturonic acid in the orange peel was converted to L-galactonic acid with a yield close to 90%. Submerged and solid-state fermentation processes were compared. PMID:24949267

2014-01-01

262

Growth Kinetics and Mechanistic Action of Reactive Oxygen Species Released by Silver Nanoparticles from Aspergillus niger on Escherichia coli  

PubMed Central

Silver Nanoparticles (AgNPs), the real silver bullet, are known to have good antibacterial properties against pathogenic microorganisms. In the present study AgNPs were prepared from extracellular filtrate of Aspergillus niger. Characterization of AgNPs by UV-Vis spectrum reveals specific surface plasmon resonance at peak 416?nm; TEM photographs revealed the size of the AgNPs to be 20–55?nm. Average diameter of the produced AgNPs was found to be 73?nm with a zeta potential that was ?24?mV using Malvern Zetasizer. SEM micrographs showed AgNPs to be spherical with smooth morphology. EDS revealed the presence of pure metallic AgNPs along with carbon and oxygen signatures. Of the different concentrations (0, 2.5, 5, 10, and 15??g/mL) used 10??g/mL were sufficient to inhibit 107?CFU/mL of E. coli. ROS production was measured using DCFH-DA method and the the free radical generation effect of AgNPs on bacterial growth inhibition was investigated by ESR spectroscopy. This paper not only deals with the damage inflicted on microorganisms by AgNPs but also induces cell death through the production of ROS released by AgNPs and also growth kinetics of E. coli supplemented with AgNPs produced by A. niger. PMID:25028666

Ninganagouda, Shivaraj; Rathod, Vandana; Singh, Dattu; Hiremath, Jyoti; Singh, Ashish Kumar; Mathew, Jasmine; ul-Haq, Manzoor

2014-01-01

263

Conversion of orange peel to L-galactonic acid in a consolidated process using engineered strains of Aspergillus niger.  

PubMed

Citrus processing waste is a leftover from the citrus processing industry and is available in large amounts. Typically, this waste is dried to produce animal feed, but sometimes it is just dumped. Its main component is the peel, which consists mostly of pectin, with D-galacturonic acid as the main monomer. Aspergillus niger is a filamentous fungus that efficiently produces pectinases for the hydrolysis of pectin and uses the resulting D-galacturonic acid and most of the other components of citrus peel for growth. We used engineered A. niger strains that were not able to catabolise D-galacturonic acid, but instead converted it to L-galactonic acid. These strains also produced pectinases for the hydrolysis of pectin and were used for the conversion of pectin in orange peel to L-galactonic acid in a consolidated process. The D-galacturonic acid in the orange peel was converted to L-galactonic acid with a yield close to 90%. Submerged and solid-state fermentation processes were compared. PMID:24949267

Kuivanen, Joosu; Dantas, Hugo; Mojzita, Dominik; Mallmann, Edgar; Biz, Alessandra; Krieger, Nadia; Mitchell, David; Richard, Peter

2014-01-01

264

The solubilization of potassium-bearing rock powder by Aspergillus niger in small-scale batch fermentations.  

PubMed

The fungus Aspergillus niger was cultivated in culture medium with an alkaline ultramafic rock powder to evaluate the solubilization of potassium for biofertilizer production. The assays were carried out with 2 strains (CCT4355 and CCT911) in small-scale batch fermentations using 125, 500, 1000, and 2000 mL Erlenmeyer flasks, with a nominal volume of 40%, and rock powder at 0.4%, shaken at 160 r/min, incubated at 30 degrees C, and sampled every 7 days for 35 days. The amount of soluble K(+), the pH of the culture medium, and the acidity were determined. Both strains solubilized K(+) from the rock powder to the same extent (approximately 62%-70% after 35 days) in the 125 mL flasks; however, the percent solubilization decreased at higher volumetric scales. The results also indicated a difference in strain sensitivity to the increase in volumetric scales in batch fermentation. When filter-sterilized air was injected into the medium, the K(+) percent solubilization obtained after 4 days of cultivation was similar to that obtained after a 28 day period. The acid production by the fungus may be a mechanism of rock solubilization, in spite of the elevation in pH values probably caused by the increasing hydrolysis of the silicates. Both strains of A. niger are recommended for solubilizing potassium from ultramafic rocks, but it is necessary to optimize the oxygen transfer, which seemed to affect the rock solubilization at higher volumetric scales. PMID:20651859

Lopes-Assad, Maria L; Avansini, Simoni H; Rosa, Márcia M; de Carvalho, José R P; Ceccato-Antonini, Sandra R

2010-07-01

265

The 53-kDa proteolytic product of precursor starch-hydrolyzing enzyme of Aspergillus niger has Taka-amylase-like activity.  

PubMed

The 53-kDa amylase secreted by Aspergillus niger due to proteolytic processing of the precursor starch-hydrolyzing enzyme was resistant to acarbose, a potent alpha-glucosidase inhibitor. The enzyme production was induced when A. niger was grown in starch medium containing the inhibitor. Antibodies against the precursor enzyme cross-reacted with the 54-kDa Taka-amylase protein of A. oryzae. It resembled Taka-amylase in most of its properties and also hydrolyzed starch to maltose of alpha-anomeric configuration. However, it did not degrade maltotriose formed during the reaction and was not inhibited by zinc ions. PMID:17123073

Ravi-Kumar, K; Venkatesh, K S; Umesh-Kumar, S

2007-04-01

266

Purification and characterization of a chlorogenic acid hydrolase from Aspergillus niger catalysing the hydrolysis of chlorogenic acid.  

PubMed

Among 15 Aspergillus strains, Aspergillus niger BRFM 131 was selected for its high chlorogenic acid hydrolase activity. The enzyme was purified and characterized with respect to its physico-chemical and kinetic properties. Four chromatographic steps were necessary to purify the protein to homogeneity with a recovery of 2%. Km of the chlorogenic acid hydrolase was estimated to be 10 microM against chlorogenic acid as substrate. Under native conditions, the protein presented a molecular mass of 170 kDa, and SDS-PAGE analysis suggested the presence of two identical 80 kDa subunits. Isoelectric point was 6.0; pH optimum for activity was determined to be 6.0 and temperature optima to be 55 degrees C. The N-terminal sequence did not present any homology with other cinnamoyl ester hydrolases previously described suggesting the purification of a new protein. The chlorogenic acid hydrolase was used successfully for the production of caffeic acid, which possesses strong antioxidant properties, from natural substrates specially rich in chlorogenic acid like apple marc and coffee pulp. PMID:15607224

Asther, Michèle; Estrada Alvarado, Maria Isabel; Haon, Mireille; Navarro, David; Asther, Marcel; Lesage-Meessen, Laurence; Record, Eric

2005-01-12

267

Molecular characterization of atoxigenic Aspergillus flavus isolates collected in China.  

PubMed

Aspergillus flavus strains were isolated from peanut fields of Liaoning, Shandong, Hubei and Guangdong Provinces in China, and identified through phenotypic and molecular approaches. Of the 323 A. flavus strains isolated, 76 strains did not produce aflatoxins detectable by UPLC. The incidence of atoxigenic A. flavus strains decreased with increase in temperature and increased with increase in latitude in different geographical locations. Amplification of all the aflatoxin genes in the aflatoxin gene cluster in the atoxigenic isolates showed that there were 25 deletion patterns (A-Y), with 22 deletion patterns identified for the first time. Most of the atoxigenic A. flavus isolates with gene deletions (97%) had deletions in at least one of the four genes (aflT, nor-1, aflR, and hypB), indicating that these four genes could be targeted for rapid identification of atoxigenic strains. The atoxigenic isolates with gene deletions, especially the isolates with large deletions, are potential candidates for aflatoxin control. PMID:24879349

Wei, Dandan; Zhou, Lu; Selvaraj, Jonathan Nimal; Zhang, Chushu; Xing, Fuguo; Zhao, Yueju; Wang, Yan; Liu, Yang

2014-07-01

268

Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88.  

PubMed

Isolates from gardening waste compost and 38 culture collection microbes were grown on agar plates at pH 4.0 with the cutinase model substrate polycaprolactone as a carbon source. The strains showing polycaprolactone hydrolysis were cultivated in liquid at acidic pH and the cultivations were monitored by assaying the p-nitrophenyl butyrate esterase activities. Culture supernatants of four strains were analyzed for the hydrolysis of tritiated apple cutin at different pHs. Highest amounts of radioactive hydrolysis products were detected at pHs below 5. The hydrolysis of apple cutin by the culture supernatants at acidic pH was further confirmed by GC-MS analysis of the hydrolysis products. On the basis of screening, the acidic cutinase from Aspergillus niger CBS 513.88 was chosen for heterogeneous production in Pichia pastoris and for analysis of the effects of pH on activity and stability. The recombinant enzyme showed activity over a broad range of pHs with maximal activity between pH 5.0 and 6.5. Activity could be detected still at pH 3.5. PMID:23540930

Nyyssölä, Antti; Pihlajaniemi, Ville; Järvinen, Riikka; Mikander, Saara; Kontkanen, Hanna; Kruus, Kristiina; Kallio, Heikki; Buchert, Johanna

2013-04-10

269

Mechanisms for Solubilization of Various Insoluble Phosphates and Activation of Immobilized Phosphates in Different Soils by an Efficient and Salinity-Tolerant Aspergillus niger Strain An2.  

PubMed

Mechanisms for solubilization of different types of phosphates and activation of immobilized phosphates in different types of soils by an efficient fungal strain An2 were explored and evaluated in this study. An2 was isolated from a Chinese cabbage rhizosphere soil and identified as Aspergillus niger. It could fast release up to 1722, 2066, and 2356 mg L(-1) of soluble phosphorus (P) from 1 % Ca3(PO4)2, Mg3(PO4)2, and AlPO4 (Ca-P, Mg-P, and Al-P) and 215 and 179 mg L(-1) from 0.5 % FePO4 and rock phosphate (Fe-P and RP), respectively. HPLC assay demonstrated that An2 mainly secreted oxalic acid to solubilize Ca-P, Mg-P, Al-P, and Fe-P whereas secreted tartaric acid to solubilize RP. Furthermore, An2 could tolerate salinity up to 4 % NaCl without impairing its phosphate-solubilizing ability. The simulation experiments validated that An2 was able to effectively activate immobilized phosphates in general calcareous, acidic, as well as saline-alkali soils with high total P content. This study shows new insights into the mechanisms for microbial solubilization of different types of phosphates and supports the future application of strain An2 in different types of soils to effectively activate P for plants. PMID:25561059

Li, Xiaolong; Luo, Lijin; Yang, Jinshui; Li, Baozhen; Yuan, Hongli

2015-03-01

270

Application of a multi-layer packed-bed reactor to citric acid production in solid-state fermentation using Aspergillus niger  

Microsoft Academic Search

Solid-state fermentation, using the fungus Aspergillus niger, has been employed for the production of citric acid from kumara, a starch-containing root crop. A multi-layer packed-bed reactor was designed and operated in an attempt to understand mass and heat transfer during the fermentation. Although only a limited understanding was obtained, the multi-layer packed-bed reactor improved the mass transfer considerably compared with

M. Y. Lu; I. S. Maddox; J. D. Brooks

1998-01-01

271

Immobilization of epoxide hydrolase from Aspergillus niger onto DEAE-cellulose: enzymatic properties and application for the enantioselective resolution of a racemic epoxide  

Microsoft Academic Search

Recombinant epoxide hydrolase (EH) from Aspergillus niger can be a very promising tool for the resolution of various racemic epoxides by enantioselective hydrolysis. The enzyme was successfully immobilized by ionic adsorption onto DEAE-cellulose (99% yield, 70% of retention activity). The temperature for maximal activity (40°C) and the activation energy (38.8kJ\\/mol) were similar for both the immobilized and free EHs, whereas

S. Karboune; A. Archelas; R. Furstoss; J. Baratti

2005-01-01

272

Altering the substrate specificity site of Aspergillus niger PhyB shifts the pH optimum to pH 3.2  

Microsoft Academic Search

Phytases are of biotechnological importance as animal feed additives for their ability to catalyze the hydrolysis of phosphate\\u000a from phytate for absorption by simple-stomached animals, and to reduce their fecal phosphorus excretion. Aspergillus niger PhyB has high catalytic activity at low pHs around 2.5, but has little activity at the commonly observed gastric pH of young\\u000a animals (3.0–3.5). Our objective

Jeremy D. Weaver; Edward J. Mullaney; Xin Gen Lei

2007-01-01

273

The impact of dose, irradiance and growth conditions on Aspergillus niger (renamed A. brasiliensis) spores low-pressure (LP) UV inactivation.  

PubMed

The use of Aspergillus niger (A. niger) fungal spores as challenge organism for UV reactor validation studies is attractive due to their high UV-resistance and non-pathogenic nature. However A. niger spores UV dose-response was dependent upon sporulation conditions and did not follow the Bunsen-Roscoe Principle of time-dose reciprocity. Exposure to 8 h of natural sunlight for 10 consecutive days increased UV resistance when compared to spores grown solely in dark conditions. Application of 250 mJ cm(-2) at high irradiance (0.11 mW cm(-2)) resulted in a 2-log inactivation; however, at low irradiance (0.022 mW cm(-2)) a 1-log inactivation was achieved. In addition, surface electron microscopy (SEM) images revealed morphological changes between the control and UV exposed spores in contrast to other well accepted UV calibrated test organisms, which show no morphological difference with UV exposure. PMID:25723059

Taylor-Edmonds, Lizbeth; Lichi, Tovit; Rotstein-Mayer, Adi; Mamane, Hadas

2015-03-21

274

Production and characterization of in planta transiently produced polygalacturanase from Aspergillus niger and its fusions with hydrophobin or ELP tags  

PubMed Central

Background Pectinases play an important role in plant cell wall deconstruction and have potential in diverse industries such as food, wine, animal feed, textile, paper, fuel, and others. The demand for such enzymes is increasing exponentially, as are the efforts to improve their production and to implement their use in several industrial processes. The goal of this study was to examine the potential of producing polygalacturonase I from Aspergillus niger in plants and to investigate the effects of subcellular compartmentalization and protein fusions on its accumulation and activity. Results Polygalacturonase I from Aspergillus niger (AnPGI) was transiently produced in Nicotiana benthamiana by targeting it to five different cellular compartments: apoplast, endoplasmic reticulum (ER), vacuole, chloroplast and cytosol. Accumulation levels of 2.5%, 3.0%, and 1.9% of total soluble protein (TSP) were observed in the apoplast, ER, and vacuole, respectively, and specific activity was significantly higher in vacuole-targeted AnPGI compared to the same enzyme targeted to the ER or apoplast. No accumulation was found for AnPGI when targeted to the chloroplast or cytosol. Analysis of AnPGI fused with elastin-like polypeptide (ELP) revealed a significant increase in the protein accumulation level, especially when targeted to the vacuole where the protein doubles its accumulation to 3.6% of TSP, while the hydrophobin (HFBI) fusion impaired AnPGI accumulation and both tags impaired activity, albeit to different extents. The recombinant protein showed activity against polygalacturonic acid with optimum conditions at pH 5.0 and temperature from 30 to 50°C, depending on its fusion. In vivo analysis of reducing sugar content revealed a higher release of reducing sugars in plant tissue expressing recombinant AnPGI compared to wild type N. benthamiana leaves. Conclusion Our results demonstrate that subcellular compartmentalization of enzymes has an impact on both the target protein accumulation and its activity, especially in the case of proteins that undergo post-translational modifications, and should be taken into consideration when protein production strategies are designed. Using plants to produce heterologous enzymes for the degradation of a key component of the plant cell wall could reduce the cost of biomass pretreatment for the production of cellulosic biofuels. PMID:24970673

2014-01-01

275

Carob pod: A new substrate for citric acid production by Aspergillus niger  

Microsoft Academic Search

The production of citric acid from carob pod extract byA. niger in surface fermentation was investigated. A maximum citric acid concentration (85.5 g\\/L), citric acid productivity (4.07\\u000a g\\/L\\/d), specific citric acid production rate (0.18 g\\/g\\/d), and specific sugar uptake rate (0.358 g\\/g\\/d) was achieved at an\\u000a initial sugar concentration of 200 g\\/L, pH of 6.5, and a temperature of 30°C.

Triantafyllos Roukas

1998-01-01

276

Characterization of species of the Aspergillus section Nigri from corn field isolates co-infected with Aspergillus flavus/parasiticus species and the potential for ochratoxin A production.  

Technology Transfer Automated Retrieval System (TEKTRAN)

Members of the Aspergillus section Nigri, known as black-spored aspergilli, can contaminate several substrates including maize. Although some species within the group can produce plant disease symptoms such as black mold in onions and maize ear rot, the main concern with A. niger aggregate contamina...

277

Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger  

PubMed Central

Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appeared to be important in order for citric acid to accumulate. The production of citric acid and the consumed sugar was higher when using 0.1% ammonium nitrate as the best source of nitrogen. The production of citric acid increased significantly when 0.1 g/l of KH2PO4 was added to the medium of date syrup. The addition of magnesium sulfate at the rate of 0.20 g/l had a stimulating effect on the production of citric acid. Maximum production of citric acid was obtained when calcium chloride was absent. One of the most important benefits of immobilized cells is their ability and stability to produce citric acid under a repeated batch culture. Over four repeated batches, the production of citric acid production was maintained for 24 days when each cycle continued for 144 h. The results obtained in the repeated batch cultivation using date syrup confirmed that date syrup could be used as a medium for the industrial production of citric acid. PMID:23961184

Mostafa, Yasser S.; Alamri, Saad A.

2012-01-01

278

A new crystal form of proteinase A, a non-pepsin-type acid proteinase from Aspergillus niger var. macrosporus.  

PubMed

Proteinase A from Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase, whose catalytic residues and mechanism remain to be elucidated. A new form of proteinase A crystals more suitable for crystallography than that obtained previously was prepared from an ammonium sulfate solution at pH 3.5 by the hanging-drop vapor diffusion method. The space group of the crystals was P2(1)2(1)2(1), with unit cell dimensions of a = 69.75 +/- 0.06 A, b = 87.55 +/- 0.05 A, and c = 60.83 +/- 0.04 A. On the assumption of two enzyme molecules per asymmetric unit, the calculated volume to unit protein mass ratio (Vm) was 2.08 A3/Da. By assuming the specific volume to be 0.74 cm3/g, the solvent content (Vso1) was estimated to be 41%, i.e., much larger than that of the crystal form obtained previously at pH 2.0 (Vso1 = 26%). Diffraction data were collected up to a resolution higher than 1.6 A, using the Weissenberg camera for macromolecular crystallography with synchrotron radiation. PMID:8276753

Tanokura, M; Sasaki, H; Muramatsu, T; Iwata, S; Hamaya, T; Takizawa, T; Takahashi, K

1993-10-01

279

Synthesis of fructooligosaccharides from Aspergillus niger commercial inulinase immobilized in montmorillonite pretreated in pressurized propane and LPG.  

PubMed

Commercial inulinase from Aspergillus niger was immobilized in montmorillonite and then treated in pressurized propane and liquefied petroleum gas (LPG). Firstly, the effects of system pressure, exposure time, and depressurization rate, using propane and LPG, on enzymatic activity were evaluated through central composite design 2³. Residual activities of 145.1 and 148.5% were observed for LPG (30 bar, 6 h, and depressurization rate of 20 bar?min?¹) and propane (270 bar, 1 h, and depressurization rate of 100 bar?min?¹), respectively. The catalysts treated at these conditions in both fluids were then used for the production of fructooligosaccharides (FOS) using sucrose and inulin as substrates in aqueous and organic systems. The main objective of this step was to evaluate the yield and productivity in FOS, using alternatives for enhancing enzyme activity by means of pressurized fluids and also using low-cost supports for enzyme immobilization, aiming at obtaining a stable biocatalyst to be used for synthesis reactions. Yields of 18% were achieved using sucrose as substrate in aqueous medium, showing the potential of this procedure, hence suggesting a further optimization step to increase the process yield. PMID:23271628

de Oliveira Kuhn, Graciele; Rosa, Clarissa Dalla; Silva, Marceli Fernandes; Treichel, Helen; de Oliveira, Débora; Oliveira, J Vladimir

2013-02-01

280

The Transcriptomic Signature of RacA Activation and Inactivation Provides New Insights into the Morphogenetic Network of Aspergillus niger  

PubMed Central

RacA is the main Rho GTPase in Aspergillus niger regulating polarity maintenance via controlling actin dynamics. Both deletion and dominant activation of RacA (RacG18V) provoke an actin localization defect and thereby loss of polarized tip extension, resulting in frequent dichotomous branching in the ?racA strain and an apolar growing phenotype for RacG18V. In the current study the transcriptomics and physiological consequences of these morphological changes were investigated and compared with the data of the morphogenetic network model for the dichotomous branching mutant ramosa-1. This integrated approach revealed that polar tip growth is most likely orchestrated by the concerted activities of phospholipid signaling, sphingolipid signaling, TORC2 signaling, calcium signaling and CWI signaling pathways. The transcriptomic signatures and the reconstructed network model for all three morphology mutants (?racA, RacG18V, ramosa-1) imply that these pathways become integrated to bring about different physiological adaptations including changes in sterol, zinc and amino acid metabolism and changes in ion transport and protein trafficking. Finally, the fate of exocytotic (SncA) and endocytotic (AbpA, SlaB) markers in the dichotomous branching mutant ?racA was followed, demonstrating that hyperbranching does not per se result in increased protein secretion. PMID:23894378

Kwon, Min Jin; Nitsche, Benjamin M.; Arentshorst, Mark; Jørgensen, Thomas R.; Ram, Arthur F. J.; Meyer, Vera

2013-01-01

281

Immobilization of Aspergillus niger F7-02 Lipase in Polysaccharide Hydrogel Beads of Irvingia gabonensis Matrix  

PubMed Central

The potential of polysaccharide Irvingia gabonensis matrix as enzyme immobilization support was investigated. Lipase of Aspergillus niger F7-02 was immobilized by entrapment using glutaraldehyde as the cross-linking agent and stabilized in ethanolic-formaldehyde solution. The pH and temperature stability and activity yield of the immobilized enzyme were determined. Such parameters as enzyme load, bead size, number of beads, and bead reusability were also optimized. Adequate gel strength to form stabilized beads was achieved at 15.52% (w/v) Irvingia gabonensis powder, 15% (v/v) partially purified lipase, 2.5% (v/v) glutaraldehyde, and 3?:?1 (v/v) ethanolic-formaldehyde solution. There was 3.93-fold purification when the crude enzyme was partially purified in two-step purification using Imarsil and activated charcoal. Optimum lipase activity 75.3?Ug?1 was achieved in 50?mL test solution containing 15 beads of 7?mm bead size. Relative activity 80% was retained at eight repeated cycles. The immobilization process gave activity yield of 59.1% with specific activity of 12.3?Umg?1 and stabilized at optimum pH 4.5 and temperature 55°C. Thus the effectiveness and cost-efficiency of I. gabonensis as a polymer matrix for lipase immobilization have been established. PMID:25614829

Kareem, Safaradeen Olateju; Adio, Olayinka Quadri; Osho, Michael Bamitale

2014-01-01

282

Encapsulation in a sol-gel matrix of lipase from Aspergillus niger obtained by bioconversion of a novel agricultural residue.  

PubMed

Lipase from Aspergillus niger was obtained from the solid-state fermentation of a novel agroindustrial residue, pumpkin seed flour. The partially purified enzyme was encapsulated in a sol-gel matrix, resulting in an immobilization yield of 71.4 %. The optimum pH levels of the free and encapsulated enzymes were 4.0 and 3.0, respectively. The encapsulated enzyme showed greater thermal stability at temperatures of 45 and 60 °C than the free enzyme. The positive influence of the encapsulation process was observed on the thermal stability of the enzyme, since a longer half-life t 1/2 and lower deactivation constant were obtained with the encapsulated lipase when compared with the free lipase. Kinetic parameters were found to follow the Michaelis-Menten equation. The K m values indicated that the encapsulation process reduced enzyme-substrate affinity and the V max was about 31.3 % lower than that obtained with the free lipase. The operational stability was investigated, showing 50 % relative activity up to six cycles of reuse at pH 3.0 at 37 °C. Nevertheless, the production of lipase from agroindustrial residue associated with an efficient immobilization method, which promotes good catalytic properties of the enzyme, makes the process economically viable for future industrial applications. PMID:24556978

Zubiolo, Claudia; Santos, Rafaela Cristiane Andrade; Carvalho, Nayara Bezerra; Soares, Cleide Mara Faria; Lima, Alvaro Silva; de Aquino Santana, Luciana Cristina Lins

2014-09-01

283

Response surface analysis for the production of an enantioselective lipase from Aspergillus niger by solid-state fermentation.  

PubMed

The lipase produced by the Aspergillus niger strain AC-54 has been widely studied due to its enantioselectivity for racemic mixtures. This study aimed to optimize the production of this enzyme using statistical methodology. Initially a Plackett-Burman (PB) design was used to evaluate the effects of the culture medium components and the culture conditions. Twelve factors were screened: water content, glucose, yeast extract, peptone, olive oil, temperature, NaH(2)P0(4), KH(2)P0(4), MgS0(4)-7H(2)0, CaCl(2), NaCI, and MnS0(4). The screening showed that the significant factors were water content, glucose, yeast extract, peptone, NaH(2)P0(4), and KH(2)P0(4), which were optimized using response surface methodology (RSM) and a mathematical model obtained to explain the behavioral process. The best lipase activity was attained using the following conditions: water content (20%), glucose (4.8%), yeast extract (4.0%), and NaH2P04 (4.0%). The predicted lipase activity was 33.03 U/ml and the experimental data confirmed the validity of the model. The enzymatic activity was expressed as micromoles of oleic acid released per minute of reaction (micromol/min). PMID:19851729

Contesini, Fabiano Jares; da Silva, Vania Castriani Fernades; Maciel, Rafael Ferreira; de Lima, Rosemary Joana; Barros, Francisco Fábio Cavalcante; de Oliveira Carvalho, Patrícia

2009-10-01

284

Optimization of Acid Protease Production by Aspergillus niger I1 on Shrimp Peptone Using Statistical Experimental Design  

PubMed Central

Medium composition and culture conditions for the acid protease production by Aspergillus niger I1 were optimized by response surface methodology (RSM). A significant influence of temperature, KH2PO4, and initial pH on the protease production was evaluated by Plackett-Burman design (PBD). These factors were further optimized using Box-Behnken design and RSM. Under the proposed optimized conditions, the experimental protease production (183.13?U?mL?1) closely matched the yield predicted by the statistical model (172.57?U?mL?1) with R2 = 0.914. Compared with the initial M1 medium on which protease production was 43.13?U?mL?1, a successful and significant improvement by 4.25 folds was achieved in the optimized medium containing (g/L): hulled grain of wheat (HGW) 5.0; KH2PO4 1.0; NaCl 0.3; MgSO4(7H2O) 0.5; CaCl2 (7H2O) 0.4; ZnSO4 0.1; Na2HPO4 1.6; shrimp peptone (SP) 1.0. The pH was adjusted at 5 and the temperature at 30°C. More interestingly, the optimization was accomplished using two cheap and local fermentation substrates, HGW and SP, which may result in a significant reduction in the cost of medium constituents. PMID:22593695

Siala, Rayda; Frikha, Fakher; Mhamdi, Samiha; Nasri, Moncef; Sellami Kamoun, Alya

2012-01-01

285

Two components of cell-bound isopullulanase from Aspergillus niger ATCC 9642--their purification and enzymatic properties.  

PubMed

Cell-bound isopullulanase (pullulan 4-glucanohydrolase: EC 3.2.1.57, IPU) from Aspergillus niger ATCC9642 [Y. Sakano et al., Denpun Kagaku, 37, 39-41 (1990)] was separated into two active components, IPU F1 (pI = 5.0) and IPU F2 (pI = 4.9), using a Mono-P HR 5/20 column. The substrate specificity on pullulan and panose, specific activity, optimum pH, pH stability, and susceptibility to certain chemical reagents were similar between IPU F1 and IPU F2. IPU F1 and F2 had an identical N-terminal amino acid sequence, A-V-T-A-D-N-S-Q-L-L. However, IPU F1 contained more total carbohydrate (15.3%) than IPU F2 (12.4%). SDS-polyacrylamide gel electrophoresis showed that the molecular weight of IPU F1 (71,000) was greater than that of IPU F2 (69,000). After deglycosylation of IPU F1 and F2 with peptide-N-glycosidase F, the molecular weights of IPU F1 and F2 became 59,000. PMID:8987855

Aoki, H; Yopi; Padmajanti, A; Sakano, Y

1996-11-01

286

Kinetic characterization of Aspergillus niger chitinase CfcI using a HPAEC-PAD method for native chitin oligosaccharides.  

PubMed

The abundant polymer chitin can be degraded by chitinases (EC 3.2.1.14) and ?-N-acetyl-hexosaminidases (EC 3.2.1.52) to oligosaccharides and N-acetyl-glucosamine (GlcNAc) monomers. Kinetic characterization of these enzymes requires product quantification by an assay method with a low detection limit, preferably compatible with the use of native, non-labeled substrates. Here we report a quantitative HPAEC-PAD method that allows fast separation of chitin oligosaccharides (COS) ranging from (GlcNac)1-6 at detection limits of 1-3 pmol and a linear range of 5-250 pmol. Quantification under intra- and interday precision conditions was performed with 2.1-5.4% relative standard deviation (RSD) and 1.2-10.3% RSD, respectively. This method was successfully used for the determination of the kinetic parameters of the Aspergillus niger chitinase CfcI with native COS. CfcI was recently shown to release GlcNAc from the reducing end of COS, a new activity for fungal chitinases. A Carbohydrate Binding Module of family 18 (CBM18) is inserted in the CfcI catalytic domain. Site directed mutagenesis was used to assess the functionality of this CfcI-CBM18: four of its key amino acids were replaced by glycine residues, yielding CfcISYNF. Comparison of the kinetic parameters of CfcI and CfcISYNF confirmed that this CBM18 is functionally involved in catalysis. PMID:25723623

van Munster, Jolanda M; Sanders, Peter; Ten Kate, Geralt A; Dijkhuizen, Lubbert; van der Maarel, Marc J E C

2015-04-30

287

Characterization of ?-Glucosidase Produced by Aspergillus niger under Solid-State Fermentation and Partially Purified Using MANAE-Agarose  

PubMed Central

?-Glucosidase (BGL) is a hydrolytic enzyme with specificity for a wide variety of glycoside substrates, being an enzyme with a large range of biotechnological applications. However, enzyme properties can be different depending both on the microorganism and the cultivation procedure employed. Therefore, in order to explore potential biocatalytical applications of novel enzymes, their characterization is essential. In this work, a BGL synthesized by a selected strain of Aspergillus niger cultivated under solid-state fermentation (SSF) was partially purified and fully characterized in terms of optimum pH, temperature, and thermostability. The single-step purification using MANAE-agarose in a chromatographic column yielded an enzyme solution with specific activity (17.1?IU/mg protein) adequate for the characterization procedures. Electrophoresis SDS-PAGE and size-exclusion chromatography analysis resulted in an estimated molecular mass of 60?kDa. Higher enzyme activities were found in the range between 40 and 65°C and between pH 4 and 5.5, indicating an interesting characteristic for application in the hydrolysis of lignocellulosic biomass for biofuels production. Thermostability studies of purified BGL resulted in half-lives at 37°C of 56.3?h and at 50°C of 5.4?h. These results provide support for further studies of this enzyme towards revealing its potential biotechnological applications. PMID:24940510

Borges, Diogo G.; Tardioli, Paulo W.; Farinas, Cristiane S.

2014-01-01

288

Production of cellulases from Aspergillus niger NS-2 in solid state fermentation on agricultural and kitchen waste residues.  

PubMed

Various agricultural and kitchen waste residues were assessed for their ability to support the production of a complete cellulase system by Aspergillus niger NS-2 in solid state fermentation. Untreated as well as acid and base-pretreated substrates including corn cobs, carrot peelings, composite, grass, leaves, orange peelings, pineapple peelings, potato peelings, rice husk, sugarcane bagasse, saw dust, wheat bran, wheat straw, simply moistened with water, were found to be well suited for the organism's growth, producing good amounts of cellulases after 96 h without the supplementation of additional nutritional sources. Yields of cellulases were higher in alkali treated substrates as compared to acid treated and untreated substrates except in wheat bran. Of all the substrates tested, wheat bran appeared to be the best suited substrate producing appreciable yields of CMCase, FPase and ?-glucosidase at the levels of 310, 17 and 33 U/g dry substrate respectively. An evaluation of various environmental parameters demonstrated that appreciable levels of cellulases could be produced over a wide range of temperatures (20-50 °C) and pH levels (3.0-8.0) with a 1:1.5 to 1:1.75 substrate to moisture ratio. PMID:22503148

Bansal, Namita; Tewari, Rupinder; Soni, Raman; Soni, Sanjeev Kumar

2012-07-01

289

Isolation and chemical characterization of naphthoquinone metabolites of Aspergillus parvulus Smith  

Microsoft Academic Search

Although several benzoquinone and anthraquinone compounds have been isolated from Aspergillus species, only two naphthoquinone monomers have been reported thus far. Aspergillus parvulus Smith (ATCC number16911) was first investigated chemically in 1974, and five naphthalenones, along with one naphthoquinone, were isolated and characterized. Based on biosynthetic considerations, it was thought that A. parvulus might be capable of producing additional naphthoquinones

1984-01-01

290

Comparsion of Cultural and Analytical Methods for Determination of Aflatoxin Production by Mississippi Delta Aspergillus isolates  

Technology Transfer Automated Retrieval System (TEKTRAN)

This study compared cultural versus analytical methods to detect aflatoxin production by Aspergillus species. Aspergillus isolates (517) were obtained from various Mississippi Delta crops (corn, peanut, rice, cotton) and soils. Ten standard aflatoxigenic A. flavus isolates were also included in thi...

291

Degradation of Xylan to d-Xylose by Recombinant Saccharomyces cerevisiae Coexpressing the Aspergillus niger ?-Xylosidase (xlnD) and the Trichoderma reesei Xylanase II (xyn2) Genes  

PubMed Central

The ?-xylosidase-encoding xlnD gene of Aspergillus niger 90196 was amplified by the PCR technique from first-strand cDNA synthesized on mRNA isolated from the fungus. The nucleotide sequence of the cDNA fragment was verified to contain a 2,412-bp open reading frame that encodes a 804-amino-acid propeptide. The 778-amino-acid mature protein, with a putative molecular mass of 85.1 kDa, was fused in frame with the Saccharomyces cerevisiae mating factor ?1 signal peptide (MF?1s) to ensure correct posttranslational processing in yeast. The fusion protein was designated Xlo2. The recombinant ?-xylosidase showed optimum activity at 60°C and pH 3.2 and optimum stability at 50°C. The Ki(app) value for d-xylose and xylobiose for the recombinant ?-xylosidase was determined to be 8.33 and 6.41 mM, respectively. The XLO2 fusion gene and the XYN2 ?-xylanase gene from Trichoderma reesei, located on URA3-based multicopy shuttle vectors, were successfully expressed and coexpressed in the yeast Saccharomyces cerevisiae under the control of the alcohol dehydrogenase II gene (ADH2) promoter and terminator. These recombinant S. cerevisiae strains produced 1,577 nkat/ml of ?-xylanase activity when expressing only the ?-xylanase and 860 nkat/ml when coexpressing the ?-xylanase with the ?-xylosidase. The maximum ?-xylosidase activity was 5.3 nkat/ml when expressed on its own and 3.5 nkat/ml when coexpressed with the ?-xylanase. Coproduction of the ?-xylanase and ?-xylosidase enabled S. cerevisiae to degrade birchwood xylan to d-xylose. PMID:11722900

La Grange, D. C.; Pretorius, I. S.; Claeyssens, M.; van Zyl, W. H.

2001-01-01

292

Enzymatic resolution of racemic phenyloxirane by a novel epoxide hydrolase from Aspergillus niger SQ-6 and its fed-batch fermentation.  

PubMed

A microorganism with the ability to catalyze the resolution of racemic phenyloxirane was isolated and identified as Aspergillus niger SQ-6. Chiral capillary electrophoresis was successfully applied to separate both phenyloxirane and phenylethanediol. The epoxide hydrolase (EH) involved in this resolution process was (R)-stereospecific and constitutively expressed. When whole cells were used during the biotransformation process, the optimum temperature and pH for stereospecific vicinal diol production were 35 degrees C and 7.0, respectively. After a 24-h conversion, the enantiomer excess of (R)-phenylethanediol produced was found to be >99%, with a conversion rate of 56%. In fed-batch fermentations at 30 degrees C for 44 h, glycerol (20 g L(-1)) and corn steep liquor (CSL) (30 g L(-1)) were chosen as the best initial carbon and nitrogen sources, and EH production was markedly improved by pulsed feeding of sucrose (2 g L(-1) h(-1)) and continuous feeding of CSL (1 g L(-1) h(-1)) at a fermentation time of 28 h. After optimization, the maximum dry cell weight achieved was 24.5+/-0.8 g L(-1); maximum EH production was 351.2+/-13.1 U L(-1) with a specific activity of 14.3+/-0.5 U g(-1). Partially purified EH exhibited a temperature optimum at 37 degrees C and pH optimum at 7.5 in 0.1 M phosphate buffer. This study presents the first evidence for the existence of a predicted epoxide racemase, which might be important in the synthesis of epoxide intermediates. PMID:16320035

Liu, Yanbin; Sha, Qian; Wu, Sheng; Wang, Jianjun; Yang, Liu; Sun, Wanru

2006-04-01

293

Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics.  

PubMed

An acidic prolyl endoprotease from Aspergillus niger was isolated from the commercial product Brewers Clarex to evaluate its possible application in proteomics. The chromatographic purification yielded a single protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis providing an apparent molecular mass of 63 kDa and a broad peak (m/z 58,061) in linear matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) indicating the glycoprotein nature of the enzyme. Indeed, a colorimetric assessment with phenol and sulfuric acid showed the presence of neutral sugars (9% of weight). The subsequent treatment with N-glycosidase F released a variety of high-mannose type N-glycans, which were successfully detected using MALDI-TOF MS. MALDI-TOF/TOF tandem MS analysis of glycopeptides from a tryptic digest of prolyl endoprotease unraveled the identity of the N-glycosylation site in the primary structure. The data obtained also show that the enzyme is present in its processed form, i.e. without putative signal and propeptide parts. Spectrophotometric measurements demonstrated optimal activity at pH 4.0-4.5 and also high thermostability for the cleavage at the C-terminal part of proline residues. In-solution digestion of standard proteins (12-200 kDa) allowed to evaluate the cleavage specificity. The enzyme acts upon proline and alanine residues, but there is an additional minor cleavage at some other residues like Gly, Leu, Arg, Ser and Tyr. The digestion of a honeybee peptide comprising six proline residues (apidaecin 1A) led to the detection of specific peptides terminated by proline as it was confirmed by MALDI postsource decay analysis. PMID:19757411

Sebela, Marek; Rehulka, Pavel; Kábrt, Jaromír; Rehulková, Helena; Ozdian, Tomás; Raus, Martin; Franc, Vojtech; Chmelík, Josef

2009-11-01

294

Effect of low shear modeled microgravity on phenotypic and central chitin metabolism in the filamentous fungi Aspergillus niger and Penicillium chrysogenum.  

PubMed

Phenotypic and genotypic changes in Aspergillus niger and Penicillium chrysogenum, spore forming filamentous fungi, with respect to central chitin metabolism were studied under low shear modeled microgravity, normal gravity and static conditions. Low shear modeled microgravity (LSMMG) response showed a similar spore germination rate with normal gravity and static conditions. Interestingly, high ratio of multiple germ tube formation of A. niger in LSMMG condition was observed. Confocal laser scanning microscopy images of calcofluor flurophore stained A. niger and P. chrysogenum showed no significant variations between different conditions tested. Transmission electron microscopy images revealed number of mitochondria increased in P. chrysogenum in low shear modeled microgravity condition but no stress related-woronin bodies in fungal hyphae were observed. To gain additional insight into the cell wall integrity under different conditions, transcription level of a key gene involved in cell wall integrity gfaA, encoding the glutamine: fructose-6-phosphate amidotransferase enzyme, was evaluated using qRT-PCR. The transcription level showed no variation among different conditions. Overall, the results collectively indicate that the LSMMG has shown no significant stress on spore germination, mycelial growth, cell wall integrity of potentially pathogenic fungi, A. niger and P. chrysogenum. PMID:24803238

Sathishkumar, Yesupatham; Velmurugan, Natarajan; Lee, Hyun Mi; Rajagopal, Kalyanaraman; Im, Chan Ki; Lee, Yang Soo

2014-08-01

295

Mapping N-linked Glycosylation Sites in the Secretome and Whole Cells of Aspergillus niger Using Hydrazide Chemistry and Mass Spectrometry  

SciTech Connect

Protein glycosylation is known to play an essential role in both cellular functions and the secretory pathways; however, little information is available on the dynamics of glycosylated N-linked glycosites of fungi. Herein we present the first extensive mapping of glycosylated N-linked glycosites in industrial strain Aspergillus niger by applying an optimized solid phase enrichment of glycopeptide protocol using hydrazide modified magnetic beads. The enrichment protocol was initially optimized using mouse plasma and A. niger secretome samples, which was then applied to profile N-linked glycosites from both the secretome and whole cell lysates of A. niger. A total of 847 unique N-linked glycosites and 330 N-linked glycoproteins were confidently identified by LC-MS/MS. Based on gene ontology analysis, the identified N-linked glycoproteins in the whole cell lysate were primarily localized in the plasma membrane, endoplasmic reticulum, golgi apparatus, lysosome, and storage vacuoles. The identified N-linked glycoproteins are involved in a wide range of biological processes including gene regulation and signal transduction, protein folding and assembly, protein modification and carbohydrate metabolism. The extensive coverage of glycosylated N-linked glycosites along with identification of partial N-linked glycosylation in those enzymes involving in different biochemical pathways provide useful information for functional studies of N-linked glycosylation and their biotechnological applications in A. niger.

Wang, Lu; Aryal, Uma K.; Dai, Ziyu; Mason, Alisa C.; Monroe, Matthew E.; Tian, Zhixin; Zhou, Jianying; Su, Dian; Weitz, Karl K.; Liu, Tao; Camp, David G.; Smith, Richard D.; Baker, Scott E.; Qian, Weijun

2012-01-01

296

Displaying Candida antarctica lipase B on the cell surface of Aspergillus niger as a potential food-grade whole-cell catalyst.  

PubMed

Aspergillus niger is a recognized workhorse used to produce food processing enzymes because of its extraordinarily high protein-producing capacity. We have developed a new cell surface display system de novo in A. niger using expression elements from generally recognized as safe certified microorganisms. Candida antarctica lipase B (CALB), a widely used hydrolase, was fused to an endogenous cell wall mannoprotein, CwpA, and functionally displayed on the cell surface. Localization of CALB was confirmed by enzymatic assay and immunofluorescence analysis using laser scanning confocal microscopy. After induction by maltose for 45 h, the hydrolytic activity and synthesis activity of A. niger mycelium-surface displayed CALB (AN-CALB) reached 400 and 240 U/g dry cell, respectively. AN-CALB was successfully used as a whole-cell catalyst for the enzymatic production of ethyl esters from a series of fatty acids of different chain lengths and ethanol. In a solvent-free system, AN-CALB showed great synthetic activity and afforded high substrate mole conversions, which amounted to 87 % for ethyl hexanoate after 2 h, 89 % for ethyl laurate after 2 h, and 84 % for ethyl stearate after 3 h. These results suggested that CwpA can act as an efficient anchoring motif for displaying enzyme on A. niger, and AN-CALB is a robust, green, and cost-effective alternative food-grade whole-cell catalyst to commercial lipase. PMID:24519503

Pan, Zhi-You; Yang, Zhi-Ming; Pan, Li; Zheng, Sui-Ping; Han, Shuang-Yan; Lin, Ying

2014-04-01

297

Overexpression of the Aspergillus niger GatA transporter leads to preferential use of D-galacturonic acid over D-xylose  

PubMed Central

Pectin is a structural heteropolysaccharide of the primary cell walls of plants and as such is a significant fraction of agricultural waste residues that is currently insufficiently used. Its main component, D-galacturonic acid, is an attractive substrate for bioconversion. The complete metabolic pathway is present in the genome of Aspergillus niger, that is used in this study. The objective was to identify the D-galacturonic acid transporter in A. niger and to use this transporter to study D-galacturonic acid metabolism. We have functionally characterized the gene An14g04280 that encodes the D-galacturonic acid transporter in A. niger. In a mixed sugar fermentation it was found that the An14g04280 overexpression strain, in contrast to the parent control strain, has a preference for D-galacturonic acid over D-xylose as substrate. Overexpression of this transporter in A. niger resulted in a strong increase of D-galacturonic acid uptake and induction of the D-galacturonic acid reductase activity, suggesting a metabolite controlled regulation of the endogenous D-galacturonic acid catabolic pathway. PMID:25177540

2014-01-01

298

Customization of Aspergillus niger morphology through addition of talc micro particles.  

PubMed

The filamentous fungus A. niger is a widely used strain in a broad range of industrial processes from food to pharmaceutical industry. One of the most intriguing and often uncontrollable characteristics of this filamentous organism is its complex morphology. It ranges from dense spherical pellets to viscous mycelia. Various process parameters and ingredients are known to influence fungal morphology. Since optimal productivity correlates strongly with a specific morphological form, the fungal morphology often represents the bottleneck of productivity in industrial production. A straight forward and elegant approach to precisely control morphological shape is the addition of inorganic insoluble micro particles (like hydrous magnesium silicate, aluminum oxide or titanium silicate oxide) to the culture medium contributing to increased enzyme production. Since there is an obvious correlation between micro particle dependent morphology and enzyme production it is desirable to mathematically link productivity and morphological appearance. Therefore a quantitative precise and holistic morphological description is targeted. Thus, we present a method to generate and characterize micro particle dependent morphological structures and to correlate fungal morphology with productivity which possibly contributes to a better understanding of the morphogenesis of filamentous microorganisms. The recombinant strain A. niger SKAn1015 is cultivated for 72 h in a 3 L stirred tank bioreactor. By addition of talc micro particles in concentrations of 1 g/L, 3 g/L and 10 g/L prior to inoculation a variety of morphological structures is reproducibly generated. Sterile samples are taken after 24, 48 and 72 hours for determination of growth progress and activity of the produced enzyme. The formed product is the high-value enzyme ?-fructofuranosidase, an important biocatalyst for neo-sugar formation in food or pharmaceutical industry, which catalyzes among others the reaction of sucrose to glucose. Therefore, the quantification of glucose after adding sucrose implies the amount of produced ?-fructofuranosidase. Glucose quantification is made by a GOD/POD-Assay, which is modified for high-throughput analysis in 96-well micro titer plates. Fungal morphology after 72 hours is examined by microscope and characterized by digital image analysis. In doing so, particle shape factors for fungal macro morphology like Feret's diameter, projected area, perimeter, circularity, aspect ratio, roundness und solidity are calculated with the open source image processing program ImageJ. Relevant parameters are combined to a dimensionless Morphology number (Mn), which enables a comprehensive characterization of fungal morphology. The close correlation of the Morphology number and productivity are highlighted by mathematical regression. PMID:22453998

Wucherpfennig, Thomas; Lakowitz, Antonia; Driouch, Habib; Krull, Rainer; Wittmann, Christoph

2012-01-01

299

Isolation and toxigenicity of Aspergillus fumigatus from moldy silage.  

PubMed

Thirty-nine silage samples were collected from various silos on Terceira Island in the Azores. Samples were examined for the presence of total fungi, and isolates of Aspergillus fumigatus were analyzed for their ability to produce fumitremorgens B and C, fumigaclavines B and C, and gliotoxin. Thirty-four silage samples (87%) were contaminated with fungi, and A. fumigatus was isolated from 27 samples (69%). Samples that were taken from the surface of silos had significantly higher populations of both total fungi and A. fumigatus than did samples taken from the middle of silos. Analysis of 27 A. fumigatus isolates (one representing each positive sample) showed that 59.3% produced fumitremorgen B; 33.3% produced fumitremorgen C; 29.6% produced fumigaclavine B; 7.4% produced fumigaclavine C; and 11.1% produced gliotoxin. Fifty-two percent of the isolates produced multiple toxins, and 25.9% did not produce any of these toxins. Gliotoxin and fumigaclavine C were always produced in combination with other toxins. Because of the demonstrated potential of these A. fumigatus isolates to produce mycotoxins, it is important to properly construct and manage silos to prevent their contamination with A. fumigatus. PMID:12733634

dos Santos, Valentina Melo; Dorner, Joe W; Carreira, Fátima

2003-01-01

300

A new member of the DMATS superfamily from Aspergillus niger catalyzes prenylations of both tyrosine and tryptophan derivatives.  

PubMed

A putative prenyltransferase gene of the dimethylallyltryptophan synthase (DMATS) family, An13g01840, was identified in the genome sequence of Aspergillus niger. The deduced polypeptide CAK41583 consists of 465 amino acids with a calculated molecular mass of 52.7 kDa. To evaluate gene function, the coding sequence was cloned into pET28a and overexpressed in Escherichia coli. The soluble His6-fusion protein was purified to near homogeneity on Ni-NTA agarose and used for enzyme assays with diverse aromatic substrates in the presence of dimethylallyl diphosphate. HPLC analysis revealed product formation in the incubation mixtures with L-tyrosine and five derivatives thereof. Structure elucidation of the enzyme products by NMR and MS analyses confirmed O-prenylations and proved the identification of a tyrosine O-prenyltransferase (TyrPT). As in the case of SirD from Leptosphaeria maculans, TyrPT also accepted 4-amino-L-phenylalanine for an N-prenylation and L-tryptophan for a C7-prenylation. The K M values of TyrPT for L-tyrosine, L-tryptophan, and dimethylallyl diphosphate (DMAPP) were found to be 0.24, 0.19, and 0.71 mM, respectively. The k cat of L-tyrosine and L-tryptophan reactions were determined at 0.58 and 0.0053 s(-1), respectively. The results presented in this study enhance the relationship of tyrosine O- and tryptophan C7-prenyltranferases and provide meanwhile a new enzyme for production of prenylated derivatives. In comparison to the known tyrosine prenyltransferase SirD, TyrPT showed significantly higher catalytic activity for several substrates, e.g., 4-amino-L-phenylalanine as well as 4- and 5-methyl-DL-tryptophan. PMID:24970457

Fan, Aili; Chen, Huizhi; Wu, Rui; Xu, Hui; Li, Shu-Ming

2014-12-01

301

The role of carbon starvation in the induction of enzymes that degrade plant-derived carbohydrates in Aspergillus niger  

PubMed Central

Fungi are an important source of enzymes for saccharification of plant polysaccharides and production of biofuels. Understanding of the regulation and induction of expression of genes encoding these enzymes is still incomplete. To explore the induction mechanism, we analysed the response of the industrially important fungus Aspergillus niger to wheat straw, with a focus on events occurring shortly after exposure to the substrate. RNA sequencing showed that the transcriptional response after 6 h of exposure to wheat straw was very different from the response at 24 h of exposure to the same substrate. For example, less than half of the genes encoding carbohydrate active enzymes that were induced after 24 h of exposure to wheat straw, were also induced after 6 h exposure. Importantly, over a third of the genes induced after 6 h of exposure to wheat straw were also induced during 6 h of carbon starvation, indicating that carbon starvation is probably an important factor in the early response to wheat straw. The up-regulation of the expression of a high number of genes encoding CAZymes that are active on plant-derived carbohydrates during early carbon starvation suggests that these enzymes could be involved in a scouting role during starvation, releasing inducing sugars from complex plant polysaccharides. We show, using proteomics, that carbon-starved cultures indeed release CAZymes with predicted activity on plant polysaccharides. Analysis of the enzymatic activity and the reaction products, indicates that these proteins are enzymes that can degrade various plant polysaccharides to generate both known, as well as potentially new, inducers of CAZymes. PMID:24792495

van Munster, Jolanda M.; Daly, Paul; Delmas, Stéphane; Pullan, Steven T.; Blythe, Martin J.; Malla, Sunir; Kokolski, Matthew; Noltorp, Emelie C.M.; Wennberg, Kristin; Fetherston, Richard; Beniston, Richard; Yu, Xiaolan; Dupree, Paul; Archer, David B.

2014-01-01

302

The role of carbon starvation in the induction of enzymes that degrade plant-derived carbohydrates in Aspergillus niger.  

PubMed

Fungi are an important source of enzymes for saccharification of plant polysaccharides and production of biofuels. Understanding of the regulation and induction of expression of genes encoding these enzymes is still incomplete. To explore the induction mechanism, we analysed the response of the industrially important fungus Aspergillus niger to wheat straw, with a focus on events occurring shortly after exposure to the substrate. RNA sequencing showed that the transcriptional response after 6h of exposure to wheat straw was very different from the response at 24h of exposure to the same substrate. For example, less than half of the genes encoding carbohydrate active enzymes that were induced after 24h of exposure to wheat straw, were also induced after 6h exposure. Importantly, over a third of the genes induced after 6h of exposure to wheat straw were also induced during 6h of carbon starvation, indicating that carbon starvation is probably an important factor in the early response to wheat straw. The up-regulation of the expression of a high number of genes encoding CAZymes that are active on plant-derived carbohydrates during early carbon starvation suggests that these enzymes could be involved in a scouting role during starvation, releasing inducing sugars from complex plant polysaccharides. We show, using proteomics, that carbon-starved cultures indeed release CAZymes with predicted activity on plant polysaccharides. Analysis of the enzymatic activity and the reaction products, indicates that these proteins are enzymes that can degrade various plant polysaccharides to generate both known, as well as potentially new, inducers of CAZymes. PMID:24792495

van Munster, Jolanda M; Daly, Paul; Delmas, Stéphane; Pullan, Steven T; Blythe, Martin J; Malla, Sunir; Kokolski, Matthew; Noltorp, Emelie C M; Wennberg, Kristin; Fetherston, Richard; Beniston, Richard; Yu, Xiaolan; Dupree, Paul; Archer, David B

2014-11-01

303

Improving the specific activity of ?-mannanase from Aspergillus niger BK01 by structure-based rational design.  

PubMed

?-Mannanase has found various biotechnological applications because it is capable of degrading mannans into smaller sugar components. A highly potent example is the thermophilic ?-mannanase from Aspergillus niger BK01 (ManBK), which can be efficiently expressed in industrial yeast strains and is thus an attractive candidate for commercial utilizations. In order to understand the molecular mechanism, which helps in strategies to improve the enzyme's performance that would meet industrial demands, 3D-structural information is a great asset. Here, we present the 1.57Å crystal structure of ManBK. The protein adopts a typical (?/?)8 fold that resembles the other GH5 family members. Polysaccharides were subsequently modeled into the substrate binding groove to identify the residues and structural features that may be involved in the catalytic reaction. Based on the structure, rational design was conducted to engineer ManBK in an attempt to enhance its enzymatic activity. Among the 23 mutants that we constructed, the most promising Y216W showed an 18±2.7% increase in specific activity by comparison with the wild type enzyme. The optimal temperature and heat tolerance profiles of Y216W were similar to those of the wild type, manifesting a preserved thermostability. Kinetic studies showed that Y216W has higher kcat values than the wild type enzyme, suggesting a faster turnover rate of catalysis. In this study we applied rational design to ManBK by using its crystal structure as a basis and identified the Y216W mutant that shows great potentials in industrial applications. PMID:24480109

Huang, Jian-Wen; Chen, Chun-Chi; Huang, Chun-Hsiang; Huang, Ting-Yung; Wu, Tzu-Hui; Cheng, Ya-Shan; Ko, Tzu-Ping; Lin, Cheng-Yen; Liu, Je-Ruei; Guo, Rey-Ting

2014-03-01

304

Elicitation of Necrosis in Vigna unguiculata Walp. by Homogeneous Aspergillus niger Endo-Polygalacturonase and by alpha-d-Galacturonate Oligomers.  

PubMed

Endo-polygalacturonase (PG) was purified from a commercial preparation of Aspergillus niger pectinase by means of carboxymethylcellulose chromatography, preparative isoelectric focusing, and gel permeation through Sephadex G-50. The enzyme was electrophoretically homogeneous and consisted of a single polypeptide chain with a molecular weight of 33,500. The enzyme exhibited a specific activity significantly higher than those of purified polygalacturonases from phytopathogenic fungi. Galacturonate oligomers with a degree of polymerization higher than four appeared quickly as products of the enzymic hydrolysis of Napolygalacturonate. The oligomers were later degraded to di- and monogalacturonate. The homogeneous enzyme and growing mycelium of Aspergillus niger separately elicited a necrotic response in cowpea (Vigna unguiculata Walp.) pods. Heat-inactivated PG and PG inactivated with specific antibodies did not elicit necrosis, suggesting that the catalytic activity of the enzyme is necessary for its function as an elicitor. The PG-released oligosaccharides from Vigna cell wall and the galacturonides with a degree of polymerization greater than four separately elicited necrosis, whereas di- and monogalacturonate did not. PMID:16665750

Cervone, F; De Lorenzo, G; Degrà, L; Salvi, G

1987-11-01

305

NON-TOXIGENIC ASPERGILLUS FLAVUS ISOLATES FOR REDUCING AFLATOXIN IN MISSISSIPPI DELTA CORN  

Technology Transfer Automated Retrieval System (TEKTRAN)

The potential for two non-toxigenic isolates of Aspergillus flavus CT3 and K49 isolated from the Mississippi Delta to reduce aflatoxin contamination of corn was assessed in a field study. These two isolates exhibited comparable growth and aggressiveness as the toxigenic A. flavus isolate F3W4. The...

306

Impact of alg3 gene deletion on growth, development, pigment production, protein secretion, and functions of recombinant Trichoderma reesei cellobiohydrolases in Aspergillus niger  

SciTech Connect

ALG3 is a Family 58 glycosyltransferase enzyme involved in early N-linked glycan synthesis. Here, we investigated the effect of the alg3 gene disruption on growth, development, metabolism, and protein secretion in Aspergillus niger. The alg3 gene deletion resulted in a significant reduction of growth on complete (CM) and potato dextrose agar (PDA) media and a substantial reduction of spore production on CM. It also delayed spore germination in the liquid cultures of both CM and PDA media, but led to a significant accumulation of red pigment on both CM and liquid modified minimal medium (MM) supplemented with yeast extract. The relative abundance of 55 proteins of the total 190 proteins identified in the secretome was significantly different as a result of alg3 gene deletion. Comparison of a Trichoderma reesei cellobiohydrolase (Cel7A) heterologously expressed in A. niger parental and ?alg3 strains showed that the recombinant Cel7A expressed in the mutant background was smaller in size than that from the parental strains. This study suggests that ALG3 is critical for growth and development, pigment production, and protein secretion in A. niger. Functional analysis of recombinant Cel7A with aberrant glycosylation demonstrates the feasibility of this alternative approach to evaluate the role of N-linked glycosylation in glycoprotein secretion and function.

Dai, Ziyu; Aryal, Uma K.; Shukla, Anil K.; Qian, Weijun; Smith, Richard D.; Magnuson, Jon K.; Adney, William S.; Beckham, Gregg T.; Brunecky, Roman; Himmel, Michael E.; Decker, Stephen R.; Ju, Xiaohui; Zhang, Xiao; Baker, Scott E.

2013-09-25

307

Malting of barley with combinations of Lactobacillus plantarum, Aspergillus niger, Trichoderma reesei, Rhizopus oligosporus and Geotrichum candidum to enhance malt quality.  

PubMed

Good quality malt is characterised by the presence of high levels of fermentable sugars, amino acids and vitamins. To reach the starch-rich endosperm of the kernel, ?-glucan- and arabinoxylan-rich cell walls have to be degraded. ?-Glucanase is synthesized in vast quantities by the aleurone layer and scutellum during germination. Secretion of hydrolytic enzymes is often stimulated by addition of the plant hormone gibberellic acid (GA3) during germination. We have shown an enhanced ?-glucanase and ?-amylase activity in malt when germinating barley was inoculated with a combination of Lactobacillus plantarum B.S1.6 and spores of Aspergillus niger MH1, Rhizopus oligosporus MH2 or Trichoderma reesei MH3, and L. plantarum B.S1.6 combined with cell-free culture supernatants from each of these fungi. Highest malt ?-glucanase activity (414 Units/kg malt) was recorded with a combination of L. plantarum B.S1.6 and spores of A. niger MH1. Highest ?-amylase activities were recorded with a combination of L. plantarum B.S1.6 and spores of R. oligosporus MH2 (373 Ceralpha Units/g malt). Highest FAN levels were recorded when L. plantarum was inoculated in combination with spores of either R. oligosporus MH2 or T. reesei MH3 (259 and 260 ppm, respectively). This is the first study showing that cell-free culture supernatants of Aspergillus, Rhizopus and Trichoderma have a stimulating effect on ?-glucanase and ?-amylase production during malting. A combination of L. plantarum B.S1.6, and spores of A. niger MH1 and R. oligosporus MH2 may be used as starter cultures to enhance malt quality. PMID:24412956

Hattingh, M; Alexander, A; Meijering, I; van Reenen, C A; Dicks, L M T

2014-03-01

308

Aspergillus, Penicillium and Talaromyces isolated from house dust samples collected around the world  

E-print Network

Aspergillus, Penicillium and Talaromyces isolated from house dust samples collected around Penicillium species of which seven were undescribed and 18 Talaromyces species including three described here, Penicillium alfredii Visagie, Seifert & Samson, P. dunedinense Visagie, Seifert & Samson, P. infrapurpureum

Amend, Anthony S.

309

Thematic course: Biotechnology of pectinolytic enzymes as instruments for structural research of plant polysaccharides. Part I. PRODUCTION OF POLYGALACTURONASES BY FILAMENTOUS FUNGI ASPERGILLUS NIGER ACM F-1119 AND PENICILLIUM DIERCKXII ACIM F-152  

Microsoft Academic Search

A comparative study of the polygalacturonases (PGs) produced by filamentous fungi Aspergillus niger ACM F-1119 and Penicillium dierckxii ACIM F-152 was carried out. PGs of the both cultures are inducible enzymes. The sugar beet pectin was found to be the most active inducer for the both cultures, and zosteran, pectin from phanerogams of Zosteraceae family was shown also to be

Anatoly A. Shubakov; Elena A. Elkina

310

Morphological development of Aspergillus niger in submerged citric acid fermentation as a function of the spore inoculum level. Application of neural network and cluster analysis for characterization of mycelial morphology  

Microsoft Academic Search

BACKGROUND: Although the citric acid fermentation by Aspergillus niger is one of the most important industrial microbial processes and various aspects of the fermentation appear in a very large number of publications since the 1950s, the effect of the spore inoculum level on fungal morphology is a rather neglected area. The aim of the presented investigations was to quantify the

Maria Papagianni; Michael Mattey

2006-01-01

311

In Vitro Activity of the Echinocandin Antifungal Agent LY303,366 in Comparison with Itraconazole and Amphotericin B against Aspergillus spp  

Microsoft Academic Search

LY303,366 (LY) is a novel derivative of the echinocandin class of antifungal agents. The in vitro activities of LY, itraconazole (ITZ), and amphotericin B (AMB) were assessed against 60 Aspergillus isolates, including 35 isolates of A. fumigatus, eight isolates of A. terreus, eight isolates of A. flavus, eight isolates of A. niger and one isolate of A. nidulans. Four A.

KAREN L. OAKLEY; CAROLINE B. MOORE; DAVID W. DENNING

1998-01-01

312

Heavy-metal-induced Inhibition of Aspergillus niger nitrate reductase: Applications for Rapid Contaminant Detection in Aqueous Samples  

SciTech Connect

Enzyme inhibition assays have the potential to rapidly screen and identify heavy metals in environmental samples. Inhibition of nitrate reductase (NR) was examined as a method for detecting toxic metals. The activity of NR (EC 1.6.6.2) from Aspergillus niger was assayed as a function of metal concentration in the presence of Cd2+, Cr3+, Cr6+, Cu2+, Ni2+, Pb2+, and Zn2+. NR exhibited sensitivity to these metals at concentrations below 10 µM. Various buffers were screened for their ability to protect NR activity from metal inhibition, and 3-(N-morpholino) propanesulfonic acid (MOPS) was selected as the buffering system for the NR assays as it exhibited the least interference with metal inhibition, thus providing increased assay sensitivity. The hypothesis that chelating agents could prevent the inhibition of NR activity by metal ions was also tested. Results indicated that 10 mM ethylenediaminetetraacetic acid (EDTA) could protect NR activity from inhibition by Cr3+, Cu2+, Cd2+, Ni2+, and Zn2+ at concentrations below 100 µM, but that the EDTA had no effect on NR inhibition by Cr6+. An amount of 10 mM nitrilotriacetic acid (NTA) prevented NR inhibition by Cd2+, Cu2+, Ni2+, Pb2+, and Zn2+ at metal concentrations below 100 µM. However, 10 mM NTA was unable to protect the enzyme from inhibition by either Cr3+ or Cr6+. These results indicated that through specific metal chelation, a NR-based method for individually quantifying Cr3+ and Cr6+ species in aqueous solutions could be developed. The ability to restore activity to NR which been previously inhibited by exposure to 100 µM Pb2+, Cd2+, Zn2+, Cu2+, and Cr3+ was explored to determine whether NR activity could be recovered by EDTA additions for use in consecutive metal inhibition assays. The results showed NR activity could not be regained after exposure to Cr3+ or Cu2+, but did partially recover activity after Cd2+, Pb2+, and Zn2+ exposure.

Apel, William Arnold; Aiken, Abigail Marie; Peyton, Brent Michael; Petersen, James N.

2003-03-01

313

Identification of amino acid residues critical for catalysis and stability in Aspergillus niger family 1 pectin lyase A.  

PubMed Central

Site-directed-mutagenesis studies were performed on family 1 pectin lyase A (PL1A) from Aspergillus niger to gain insight into the reaction mechanism for the pectin lyase-catalysed beta-elimination cleavage of methylesterified polygalacturonic acid and to stabilize the enzyme at slightly basic pH. On the basis of the three-dimensional structures of PL1A [Mayans, Scott, Connerton, Gravesen, Benen, Visser, Pickersgill and Jenkins (1997) Structure 5, 677-689] and the modelled enzyme-substrate complex of PL1B [Herron, Benen, Scavetta, Visser and Jurnak (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 8762-8769], Asp154, Arg176, Arg236 and Lys239 were mutagenized. Substituting Arg236 with alanine or lysine rendered the enzyme completely inactive, and mutagenesis of Arg176 and Lys239 severely affected catalysis. The Asp154-->Arg and Asp154-->Glu mutant enzymes were only moderately impaired in respect of catalysis. The results strongly indicate that Arg236, which is sandwiched between Arg176 and Lys239, would initiate the reaction upon enzyme-substrate interaction, through the abstraction of the proton at C5 of the galacturonopyranose ring. The positively charged residues Arg176 and Lys239 are responsible for lowering the p K a of Arg236. Arg176 and Lys239 are maintained in a charged state by interacting with Asp154 or bulk solvent respectively. The deprotonation of the Asp186-Asp221 pair was proposed to be responsible for a pH-driven conformational change of PL1A [Mayans, Scott, Connerton, Gravesen, Benen, Visser, Pickersgill and Jenkins (1997) Structure 5, 677-689]. Substitution of Asp186 and Asp221 by Asn186 and Asn221 was expected to stabilize the enzyme. However, the Asp186-->Asn/Asp221-->Asn enzyme appeared less stable than the wild-type enzyme, even at pH 6.0, as evidenced by fluorescence studies. This demonstrates that the pH-dependent conformational change is not driven by deprotonation of the Asp186-Asp221 pair. PMID:12418964

Sánchez-Torres, Paloma; Visser, Jaap; Benen, Jacques A E

2003-01-01

314

Multiple Genetically Distinct Groups Revealed among Clinical Isolates Identified as Atypical Aspergillus fumigatus  

Microsoft Academic Search

To investigate whether genetic variants of A. fumigatus are found among clinical isolates, four isolates that were originally identified as poorly sporulating strains of Aspergillus fumigatus were subjected to molecular analysis. DNA sequence analysis of the alkaline protease genes of these isolates showed that each is genetically distinct and each shows substantial variation (7 to 11%) from the A. fumigatus

Margaret E. Katz; Annette M. Dougall; Brian F. Cheetham

2005-01-01

315

Relationships between aflatoxin production, sclerotia formation among isolates of Aspergillus section Flavi from the Mississippi Delta  

Technology Transfer Automated Retrieval System (TEKTRAN)

Aspergillus isolates from different crops and from soils of the Mississippi Delta differed significantly in production of aflatoxin and sclerotia. Overall about 50% of the isolates from corn, soil, and peanut produced large sclerotia, while only 20% of the rice isolates produced large sclerotia. T...

316

Isolation of Two Apsa Suppressor Strains in Aspergillus Nidulans  

PubMed Central

Aspergillus nidulans reproduces asexually with single nucleated conidia. In apsA (anucleate primary sterigmata) strains, nuclear positioning is affected and conidiation is greatly reduced. To get further insights into the cellular functions of apsA, aconidial apsA strains were mutagenized and conidiating suppressor strains were isolated. The suppressors fell into two complementation groups, samA and samB (suppressor of anucleate metulae). samA mapped on linkage group I close to pyrG. The mutant allele was dominant in diploids homozygous for apsA. Viability of conidia of samA suppressor strains (samA(-); apsA(-)) was reduced to 50% in comparison to wild-type conidia. Eighty percent of viable spores produced small size colonies that were temperature- and benomyl-sensitive. samB mapped to chromosome VIII and was recessive. Viability of conidia from samB suppressor strains (apsA(-); samB(-)) was also affected but no small size colonies were observed. Both suppressors produced partial defects in sexual reproduction and both suppressed an apsA deletion mutation. In wild-type background the mutant loci affected hyphal growth rate (samA) or changed the colony morphology (samB) and inhibited sexual spore formation (samA and samB). Only subtle effects on conidiation were found. We conclude that both suppressor genes bypass the apsA function and are involved in microtubule-dependent processes. PMID:8889518

Kruger, M.; Fischer, R.

1996-01-01

317

Utilisation of plant cell-wall polysaccharides and organic phosphorus substrates by isolates of Aspergillus and Penicillium isolated from soil  

Microsoft Academic Search

The ability of seventy isolates (comprising 43 species) of Aspergillus and Penicillium, from soil and compost, to grow on sources of carbon and phosphate from plant remains was examined. Only two isolates from compost actively degraded crystalline cellulose, though most others grew on carboxymethyl cellulose. Most isolates produced biomass on cellobiose, and all on glucose, pectin and xylan. All fungi

Cathal M. Daynes; Peter A. McGee; David J. Midgley

2008-01-01

318

Morphological and molecular identification of filamentous Aspergillus flavus and Aspergillus parasiticus isolated from compound feeds in South Africa.  

PubMed

Isolation of filamentous species of two Aspergillum genera from compound feeds produced in South Africa, and subsequent extraction of their individual DNA in this study, presents a simple but rapid molecular procedure for high through-put analysis of the individual morphological forms. DNA was successfully isolated from the Aspergillus spp. from agar cultures by use of a commercial kit. Agarose gel electrophoresis fractionation of the fungi DNA, showed distinct bands. The DNA extracted by this procedure appears to be relatively pure with a ratio absorbance at 260 and 280 nm. However, the overall morphological and molecular data indicated that 67.5 and 51.1% of feed samples were found to be contaminated with Aspergillus flavus and Aspergillus parasiticus, respectively, with poultry feed having the highest contamination mean level of 5.7 × 105 CFU/g when compared to cattle (mean: 4.0 × 106 CFU/g), pig (mean: 2.7 × 104 CFU/g) and horse (1.0 × 102 CFU) feed. This technique presents a readily achievable, easy to use method in the extraction of filamentous fungal DNA and it's identification. Hence serves as an important tool towards molecular study of these organisms for routine analysis check in monitoring and improving compound feed quality against fungal contamination. PMID:25084661

Iheanacho, Henry E; Njobeh, Patrick B; Dutton, Francis M; Steenkamp, Paul A; Steenkamp, Lucia; Mthombeni, Julian Q; Daru, Barnabas H; Makun, Anthony H

2014-12-01

319

Aminopeptidases from Aspergillus niger  

Microsoft Academic Search

Aspergillusis a filamentous fungus that can grow in many environments, on several substrates at different conditions. In the soil,Aspergilli<\\/span>recycle nutrients by the degradation of plant material. In particular,Aspergilli<\\/span>are known for their capability to spoil plant materials like grains and fruits. In the industryAspergilli<\\/span>are often being used as hosts for the production of both homologous andheterologous<\\/span>(foodgrade<\\/span>)

Wijk van D

2004-01-01

320

Water relations of Paecilomyces variotii, Eurotium amstelodami, Aspergillus candidus and Aspergillus sydowii, xerophilic fungi isolated from Indonesian dried fish.  

PubMed

The water relations of four xerotolerant fungi, Paecilomyces variotii, Eurotium amstelodami, Aspergillus candidus and Aspergillus sydowii, isolated from dried salt fish, were examined at 25 degrees C, on media in which water activity (aW) was controlled by NaCl or a glucose/fructose mixture. All fungi were less tolerant of NaCl than glucose/fructose at low aW. P. variotii grew 2 to 3 times faster on glucose/fructose media than on NaCl. The minimum aW permitting germination varied from 0.753 for E. amstelodami and, 0.776 for A. candidus and A. sydowii to 0.793 for P. variotii. At low aW germination was not always followed by growth. In most cases the minimum for growth was 0.02 aW units above that for germination. PMID:3275312

Wheeler, K A; Hocking, A D

1988-08-01

321

Characterization of the starvation-induced chitinase CfcA and ?-1,3-glucanase AgnB of Aspergillus niger.  

PubMed

The common saprophyte Aspergillus niger may experience carbon starvation in nature as well as during industrial fermentations. Starvation survival strategies, such as conidiation or the formation of exploratory hyphae, require energy and building blocks, which may be supplied by autolysis. Glycoside hydrolases are key effectors of autolytic degradation of fungal cell walls, but knowledge on their identity and functionality is still limited. We recently identified agnB and cfcA as two genes encoding carbohydrate-active enzymes that had notably increased transcription during carbon starvation in A. niger. Here, we report the biochemical and functional characterization of these enzymes. AgnB is an ?-1,3-glucanase that releases glucose from ?-1,3-glucan substrates with a minimum degree of polymerization of 4. CfcA is a chitinase that releases dimers from the nonreducing end of chitin. These enzymes thus attack polymers that are found in the fungal cell wall and may have a role in autolytic fungal cell wall degradation in A. niger. Indeed, cell wall degradation during carbon starvation was reduced in the double deletion mutant ?cfcA ?agnB compared to the wild-type strain. Furthermore, the cell walls of the carbon-starved mycelium of the mutant contained a higher fraction of chitin or chitosan. The function of at least one of these enzymes, CfcA, therefore appears to be in the recycling of cell wall carbohydrates under carbon limiting conditions. CfcA thus may be a candidate effector for on demand cell lysis, which could be employed in industrial processes for recovery of intracellular products. PMID:25219534

van Munster, Jolanda M; Dobruchowska, Justyna M; Veloo, Ruud; Dijkhuizen, Lubbert; van der Maarel, Marc J E C

2015-03-01

322

Isolation of maize soil and rhizosphere bacteria with antagonistic activity against Aspergillus flavus and Fusarium verticillioides  

Technology Transfer Automated Retrieval System (TEKTRAN)

Bacterial isolates from Mississippi maize field soil and maize rhizosphere samples were evaluated for their potential as biological control agents against Aspergillus flavus and Fusarium verticillioides. Isolated strains were screened for antagonistic activities in liquid co-culture against A. flav...

323

ENZYMATIC DEHAIRING OF CATTLE HIDE WITH AN ALKALINE PROTEASE ISOLATED FROM ASPERGILLUS TAMARII  

Technology Transfer Automated Retrieval System (TEKTRAN)

An alkaline protease isolated from Aspergillus tamarii shows promise as a dehairing agent for use in the tannery. Standard dehairing conditions established for the protease isolated from Streptomyces griseus proved to be unsatisfactory for the alkaline protease. We optimized the dehairing conditio...

324

Aflaquinolones A-G: Secondary metabolites from marine and fungicolous isolates of Aspergillus spp.  

Technology Transfer Automated Retrieval System (TEKTRAN)

Seven new compounds (aflaquinolones A-G; 1-7) containing dihydroquinolin-2-one and terpenoid units have been isolated from two different fungal sources. Two of these metabolites (1 and 2) were obtained from a Hawaiian fungicolous isolate of Aspergillus sp. (section Flavipedes; MYC-2048=NRRL 58570), ...

325

Comparison of Aflatoxigenic and Nonaflatoxigenic Isolates of Aspergillus flavus using DNA Amplification Fingerprinting Techniques  

Microsoft Academic Search

Aspergillus flavus is a filamentous fungus that produces mycotoxins in many food and feed crops, such as maize (Zea mays L.). Isolates were analyzed for toxin production by nucleic acid profiles in an attempt to differentiate aflatoxigenic from\\u000a nonaflatoxigenic isolates. A total of 41 aflatoxigenic and 34 nonalfatoxigenic isolates were included in the study. The isolates\\u000a were evaluated initially using

R. E. Baird; R. N. Trigiano; G. Windham; P. Williams; R. Kelley; H. K. Abbas; J. K. Moulton; M. L. Scruggs

2006-01-01

326

Expression and secretion in Aspergillus nidulans and Aspergillus niger of a cell surface glycoprotein from the cattle tick, Boophilus microplus, by using the fungal amdS promoter system.  

PubMed Central

A cell surface glycoprotein (Bm86) from cells of the digestive tract of the cattle tick Boophilus microplus, which has been shown to elicit a protective immunological response in vaccinated cattle, was expressed and secreted in the filamentous fungi Aspergillus nidulans and Aspergillus niger by using the fungal amdS promoter system. The cloned gene coded for the Bm86 secretory signal and all of the Bm86 mature polypeptide except for the hydrophobic carboxy-terminal segment. High levels of Bm86 mRNA were detected in the transformed cells. Bm86 polypeptide was secreted from the cells in a soluble form and it was glycosylated, probably to a similar extent to the native glycoprotein. The recombinant product had an apparent molecular mass of 83 to 87 kilodaltons, whereas that predicted from the amino acid sequence was 69 kilodaltons. The Bm86 was expressed at levels of up to 1.8 mg/liter, or approximately 6% of secreted protein under the growth conditions used. No intracellular Bm86 was detected. A general relationship was observed between transformants containing a high number of copies of the expression plasmid and high expression levels. Images PMID:2275533

Turnbull, I F; Smith, D R; Sharp, P J; Cobon, G S; Hynes, M J

1990-01-01

327

Biodiversity and ITS-RFLP Characterisation of Aspergillus Section Nigri Isolates in Grapes from Four Traditional Grape-Producing Areas in Greece  

PubMed Central

A study on the occurrence of Aspergillus section Nigri species on grapes from four traditional grape-producing areas in Greece during the 2011/2012 vintage, and their capability to produce OTA was conducted. One hundred and twenty-eight black aspergilli isolates were characterised at the species level initially by the use of morphological criteria in accordance with appropriate keys, followed by molecular characterisation performed with Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR-RFLP) of the 5.8 ribosomal RNA gene Internal Transcribed Spacer region (5.8 rRNA ITS). Restriction enzyme digestion of the ITS amplicons using the HhaI, HinfI and RsaI, endonucleases distinguished eleven different patterns of restriction fragment length polymorphism (RFLP), four for each of the HhaI and RsaI digests and three for HinfI. From a total number of 128 individual isolates, 124 were classified into four Aspergillus species corresponding to A. carbonarius, A. tubingensis, A. japonicus and A. ibericus, and the remaining 4 were classified as members of the A. niger aggregate. A. carbonarius and A. tubingensis being the main representative species were equally counted, with higher geographical representation of the former in southern and the latter in northern regions, respectively. All isolates were tested for their ochratoxigenic potential by use of High Performance Liquid Chromatography (HPLC) and Enzyme Linked Immuno Sorbent Assay (ELISA), resulting in significant interspecies differences in OTA production. PMID:24710283

Kizis, Dimosthenis; Natskoulis, Pantelis; Nychas, George-John E.; Panagou, Efstathios Z.

2014-01-01

328

The effectiveness of arbuscular-mycorrhizal fungi and Aspergillus niger or Phanerochaete chrysosporium treated organic amendments from olive residues upon plant growth in a semi-arid degraded soil  

Microsoft Academic Search

Arbuscular mycorrhizal (AM) fungi and a residue from dry olive cake (DOC) supplemented with rock phosphate (RP) and treated with either Aspergillus niger (DOC-A) or Phanerochaete chrysosporium (DOC-P), were assayed in a natural, semi-arid soil using Trifolium repens or Dorycnium pentaphyllum plants. The effects of the AM fungi and\\/or DOC-A were compared with P-fertilisation (P) over eleven successive harvests to evaluate

A. Medina; A. Roldán; R. Azcón

2010-01-01

329

Aspergillus sp. isolated in critically ill patients with extracorporeal membrane oxygenation support.  

PubMed

This study reports Aspergillus isolation in critically ill patients who underwent extracorporeal membrane oxygenation (ECMO) and highlights the difficulty in establishing a diagnosis of aspergillosis in this population. The diagnosis of Aspergillus infection or colonization was retrospectively performed using the proposed modified criteria of the European Organization for Research and Treatment of Cancer and the Mycoses Study Group (EORTC/MSG) adapted to critically ill patients. Between 2005 and 2011, 11 of 151 patients (7.2%) who underwent ECMO had Aspergillus sp. isolates, 10 in a pulmonary sample and 1 in a mediastinal wound sample. Five patients did not have any classical risk factors for aspergillosis. One patient had a proven invasive pulmonary aspergillosis (IPA), 2 had a putative IPA, and 1 patient had a possible Aspergillus mediastinitis, whilst in 7 patients this was considered colonization. However, the clinical relevance of Aspergillus isolation was based on an algorithm not validated in patients undergoing ECMO. Our data support the need to implement non-invasive diagnostic procedures for aspergillosis in this population. PMID:23746344

Aubron, Cecile; Pilcher, David; Leong, Tim; Cooper, D James; Scheinkestel, Carlos; Pellegrino, Vince; Cheng, Allen C

2013-09-01

330

Simultaneous purification and immobilization of Aspergillus niger xylanase on the reversibly soluble polymer Eudragit TM L-100  

Microsoft Academic Search

The non-covalent immobilization of a commercial preparation of xylanase from A. niger was carried out on a reversibly soluble-insoluble enteric polymer EudragitTM L-100. The immobilization of the xylanase activity by adsorption was simultaneously accompanied by removal of cellulase activity since the latter did not bind to the polymer. Thus, the soluble enzyme derivative may be useful for treatment of paper

Meryam Sardar; Ipsita Roy; Munishwar N Gupta

2000-01-01

331

The Transcriptional Repressor TupA in Aspergillus niger Is Involved in Controlling Gene Expression Related to Cell Wall Biosynthesis, Development, and Nitrogen Source Availability  

PubMed Central

The Tup1-Cyc8 (Ssn6) complex is a well characterized and conserved general transcriptional repressor complex in eukaryotic cells. Here, we report the identification of the Tup1 (TupA) homolog in the filamentous fungus Aspergillus niger in a genetic screen for mutants with a constitutive expression of the agsA gene. The agsA gene encodes a putative alpha-glucan synthase, which is induced in response to cell wall stress in A. niger. Apart from the constitutive expression of agsA, the selected mutant was also found to produce an unknown pigment at high temperatures. Complementation analysis with a genomic library showed that the tupA gene could complement the phenotypes of the mutant. Screening of a collection of 240 mutants with constitutive expression of agsA identified sixteen additional pigment-secreting mutants, which were all mutated in the tupA gene. The phenotypes of the tupA mutants were very similar to the phenotypes of a tupA deletion strain. Further analysis of the tupA-17 mutant and the ?tupA mutant revealed that TupA is also required for normal growth and morphogenesis. The production of the pigment at 37°C is nitrogen source-dependent and repressed by ammonium. Genome-wide expression analysis of the tupA mutant during exponential growth revealed derepression of a large group of diverse genes, including genes related to development and cell wall biosynthesis, and also protease-encoding genes that are normally repressed by ammonium. Comparison of the transcriptome of up-regulated genes in the tupA mutant showed limited overlap with the transcriptome of caspofungin-induced cell wall stress-related genes, suggesting that TupA is not a general suppressor of cell wall stress-induced genes. We propose that TupA is an important repressor of genes related to development and nitrogen metabolism. PMID:24205111

Schachtschabel, Doreen; Arentshorst, Mark; Nitsche, Benjamin M.; Morris, Sam; Nielsen, Kristian F.; van den Hondel, Cees A. M. J. J.; Klis, Frans M.; Ram, Arthur F. J.

2013-01-01

332

Systems Approaches to Predict the Functions of Glycoside Hydrolases during the Life Cycle of Aspergillus niger Using Developmental Mutants ?brlA and ?flbA  

PubMed Central

Background The filamentous fungus Aspergillus niger encounters carbon starvation in nature as well as during industrial fermentations. In response, regulatory networks initiate and control autolysis and sporulation. Carbohydrate-active enzymes play an important role in these processes, for example by modifying cell walls during spore cell wall biogenesis or in cell wall degradation connected to autolysis. Results In this study, we used developmental mutants (?flbA and ?brlA) which are characterized by an aconidial phenotype when grown on a plate, but also in bioreactor-controlled submerged cultivations during carbon starvation. By comparing the transcriptomes, proteomes, enzyme activities and the fungal cell wall compositions of a wild type A. niger strain and these developmental mutants during carbon starvation, a global overview of the function of carbohydrate-active enzymes is provided. Seven genes encoding carbohydrate-active enzymes, including cfcA, were expressed during starvation in all strains; they may encode enzymes involved in cell wall recycling. Genes expressed in the wild-type during starvation, but not in the developmental mutants are likely involved in conidiogenesis. Eighteen of such genes were identified, including characterized sporulation-specific chitinases and An15g02350, member of the recently identified carbohydrate-active enzyme family AA11. Eight of the eighteen genes were also expressed, independent of FlbA or BrlA, in vegetative mycelium, indicating that they also have a role during vegetative growth. The ?flbA strain had a reduced specific growth rate, an increased chitin content of the cell wall and specific expression of genes that are induced in response to cell wall stress, indicating that integrity of the cell wall of strain ?flbA is reduced. Conclusion The combination of the developmental mutants ?flbA and ?brlA resulted in the identification of enzymes involved in cell wall recycling and sporulation-specific cell wall modification, which contributes to understanding cell wall remodeling mechanisms during development. PMID:25629352

van Munster, Jolanda M.; Nitsche, Benjamin M.; Akeroyd, Michiel; Dijkhuizen, Lubbert; van der Maarel, Marc J. E. C.; Ram, Arthur F. J.

2015-01-01

333

Development of cost-effective media to increase the economic potential for larger-scale bioproduction of natural food additives by Lactobacillus rhamnosus , Debaryomyces hansenii , and Aspergillus niger.  

PubMed

Yeast extract (YE) is the most common nitrogen source in a variety of bioprocesses in spite of the high cost. Therefore, the use of YE in culture media is one of the major technical hurdles to be overcome for the development of low-cost fermentation routes, making the search for alternative-cheaper nitrogen sources particularly desired. The aim of the current study is to develop cost-effective media based on corn steep liquor (CSL) and locally available vinasses in order to increase the economic potential for larger-scale bioproduction. Three microorganisms were evaluated: Lactobacillus rhamnosus , Debaryomyces hansenii , and Aspergillus niger . The amino acid profile and protein concentration was relevant for the xylitol and citric acid production by D. hansenii and A. niger , respectively. Metals also played an important role for citric acid production, meanwhile, D. hansenii showed a strong dependence with the initial amount of Mg(2+). Under the best conditions, 28.8 g lactic acid/L (Q(LA) = 0.800 g/L.h, Y(LA/S) = 0.95 g/g), 35.3 g xylitol/L (Q(xylitol) = 0.380 g/L.h, Y(xylitol/S) = 0.69 g/g), and 13.9 g citric acid/L (Q(CA) = 0.146 g/L.h, Y(CA/S) = 0.63 g/g) were obtained. The economic efficiency (E(p/euro)) parameter identify vinasses as a lower cost and more effective nutrient source in comparison to CSL. PMID:19821581

Salgado, José Manuel; Rodríguez, Noelia; Cortés, Sandra; Domínguez, José Manuel

2009-11-11

334

Penicillium parvulum and Penicillium georgiense sp. nov. Isolated from the Conidial Heads of Aspergillus Species  

Technology Transfer Automated Retrieval System (TEKTRAN)

Two new Penicillium species were isolated from peanut-field soils in Georgia. The species were particularly noted because they sporulated on the conidial heads of Aspergillus species. Phenotypic descriptions were prepared using standard media. ITS and lsu-rDNA sequences were made from the new spe...

335

Enzymatic Dehairing of Cattlehide with an Alkaline Protease Isolated from Aspergillus tamarii  

Technology Transfer Automated Retrieval System (TEKTRAN)

An enzymatic dehairing protocol based on the alkaline serine protease isolated from Aspergillus tamarii required 16h, and we observed concomitant grain damage. The use of sodium dodecyl sulfate as a pretreatment to remove the lipids from the hide allowed a shortening of the dehairing time to 6 h wi...

336

A Potent Nonpeptide Cholecystokinin Antagonist Selective for Peripheral Tissues Isolated from Aspergillus alliaceus  

Microsoft Academic Search

A new, competitive, nonpeptide cholecystokinin (CCK) antagonist, asperlicin, was isolated from the fungus Aspergillus alliaceus. The compound has 300 to 400 times the affinity for pancreatic, ileal, and gallbladder CCK receptors than proglumide, a standard agent of this class. Moreover, asperlicin is highly selective for peripheral CCK receptors relative to brain CCK and gastrin receptors. Since asperlicin also exhibits long-lasting

Raymond S. L. Chang; Victor J. Lotti; Richard L. Monaghan; Jerome Birnbaum; Edward O. Stapley; Michael A. Goetz; Georg Albers-Schonberg; Arthur A. Patchett; Jerrold M. Liesch; Otto D. Hensens; James P. Springer

1985-01-01

337

Aspergillus, Penicillium and Talaromyces isolated from house dust samples collected around the world  

PubMed Central

As part of a worldwide survey of the indoor mycobiota, dust was collected from nine countries. Analyses of dust samples included the culture-dependent dilution-to-extinction method and the culture-independent 454-pyrosequencing. Of the 7?904 isolates, 2?717 isolates were identified as belonging to Aspergillus, Penicillium and Talaromyces. The aim of this study was to identify isolates to species level and describe the new species found. Secondly, we wanted to create a reliable reference sequence database to be used for next-generation sequencing projects. Isolates represented 59 Aspergillus species, including eight undescribed species, 49 Penicillium species of which seven were undescribed and 18 Talaromyces species including three described here as new. In total, 568 ITS barcodes were generated, and 391 ?-tubulin and 507 calmodulin sequences, which serve as alternative identification markers. PMID:25492981

Visagie, C.M.; Hirooka, Y.; Tanney, J.B.; Whitfield, E.; Mwange, K.; Meijer, M.; Amend, A.S.; Seifert, K.A.; Samson, R.A.

2014-01-01

338

Aflaquinolones A-G: secondary metabolites from marine and fungicolous isolates of Aspergillus spp.  

PubMed

Seven new compounds (aflaquinolones A-G; 1-7) containing dihydroquinolin-2-one and terpenoid units have been isolated from two different fungal sources. Two of these metabolites (1 and 2) were obtained from a Hawaiian fungicolous isolate of Aspergillus sp. (section Flavipedes; MYC-2048 = NRRL 58570), while the others were obtained from a marine Aspergillus isolate (SF-5044) collected in Korea. The structures of these compounds were determined mainly by analysis of NMR and MS data. Relative and absolute configurations were assigned on the basis of NOESY data and (1)H NMR J-values, comparison of calculated and experimental ECD spectra, and analysis of a Mosher's ester derivative of 2. Several known compounds, including alantrypinone, aspochalasins I and J, methyl 3,4,5-trimethoxy-2((2-((3-pyridinylcarbonyl)amino)benzoyl)amino)benzoate, and trans-dehydrocurvularin were also encountered in the extract of the Hawaiian isolate. PMID:22295903

Neff, Scott A; Lee, Sang Un; Asami, Yukihiro; Ahn, Jong Seog; Oh, Hyuncheol; Baltrusaitis, Jonas; Gloer, James B; Wicklow, Donald T

2012-03-23

339

HYOSCYAMINE 6 ? ?-HYDROXYLASE GENE ISOLATION FROM IN VITRO CULTURED ROOTS OF HYOSCYAMUS NIGER L. AND HYOSCAYMUS TENUICAULIS SCHONBECK-TEMESY  

Microsoft Academic Search

The H6H gene for hyoscyamine 6?-hydroxylase, which converts hyoscyamine to scopolamine, was isolated from H. niger and H. tenuicaulis. The roots of 14 days sterile seedlings were transferred to a modified liquid B5 medium containing 1? M in dolebutyric acid, and after appearance of the lateral roots, subcultured in a free hormone medium. Following a week, the total cellular RNA

HASSAN EBRAHIMZADEH; AFSANEH TEIMOORI; TAHMINEH LOHRASEBI

2003-01-01

340

New cytotoxic sesquiterpenoid nitrobenzoyl esters from a marine isolate of the fungus Aspergillus versicolor  

Microsoft Academic Search

Four new sesquiterpenoid nitrobenzoyl esters (1–4) have been isolated from organic extracts of the culture broth and mycelia of Aspergillus versicolor, a fungus isolated from the surface of the Caribbean green alga Penicillus capitatus. The structures of the four compounds were determined through extensive analysis of 1H NMR, 13C NMR, HMQC, and HMBC data. 9?, 14-Dihydroxy-6?-p-nitrobenzoylcinnamolide (1) displayed significant cytotoxicity

Gilbert N. Belofsky; Paul R. Jensen; Matthew K. Renner; William Fenical

1998-01-01

341

Distribution and toxigenicity of Aspergillus species isolated from maize kernels from three agro-ecological zones in Nigeria  

E-print Network

Distribution and toxigenicity of Aspergillus species isolated from maize kernels from three agro agro-ecological zones in Nigeria to determine the distribution and aflatoxin-producing potential of members of Aspergillus section Flavi. The three agro-ecological zones were, Derived Savannah (DS

Cotty, Peter J.

342

Crystallization and preliminary X-ray investigation of proteinase A, a non-pepsin-type acid proteinase from Aspergillus niger var. macrosporus.  

PubMed

Proteinase A from Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase distinctly different in various properties from the family of pepsin-type aspartic proteinases, and so far it remains unknown which residues participate in the catalysis of the enzyme and how the mechanism operates. The acid proteinase A was crystallized from an ammonium sulfate solution by the hanging-drop vapor diffusion method. The space group of the crystals was P2(1)2(1)2(1) with unit cell dimensions of a = 54.7 A, b = 70.4 A and c = 38.0 A. On the assumption that there is one enzyme molecule in the asymmetric unit, the calculated ratio of volume to unit protein mass (Vm) was 1.64 A3 per dalton. Diffraction data were collected up to a resolution higher than 1.5 A, using the Weissenberg camera for macromolecular crystallography with synchrotron radiation. The crystal of proteinase A is, therefore, suitable for the structural analysis with a high resolution. PMID:1731082

Tanokura, M; Matsuzaki, H; Iwata, S; Nakagawa, A; Hamaya, T; Takizawa, T; Takahashi, K

1992-01-01

343

Effects of High Pressure Homogenization on the Activity, Stability, Kinetics and Three-Dimensional Conformation of a Glucose Oxidase Produced by Aspergillus niger  

PubMed Central

High pressure homogenization (HPH) is a non-thermal method, which has been employed to change the activity and stability of biotechnologically relevant enzymes. This work investigated how HPH affects the structural and functional characteristics of a glucose oxidase (GO) from Aspergillus niger. The enzyme was homogenized at 75 and 150 MPa and the effects were evaluated with respect to the enzyme activity, stability, kinetic parameters and molecular structure. The enzyme showed a pH-dependent response to the HPH treatment, with reduction or maintenance of activity at pH 4.5–6.0 and a remarkable activity increase (30–300%) at pH 6.5 in all tested temperatures (15, 50 and 75°C). The enzyme thermal tolerance was reduced due to HPH treatment and the storage for 24 h at high temperatures (50 and 75°C) also caused a reduction of activity. Interestingly, at lower temperatures (15°C) the activity levels were slightly higher than that observed for native enzyme or at least maintained. These effects of HPH treatment on function and stability of GO were further investigated by spectroscopic methods. Both fluorescence and circular dichroism revealed conformational changes in the molecular structure of the enzyme that might be associated with the distinct functional and stability behavior of GO. PMID:25061935

Tribst, Alline Artigiani Lima; Cota, Júnio; Murakami, Mario Tyago; Cristianini, Marcelo

2014-01-01

344

Effects of elevated dissolved CO2 levels on batch and continuous cultures of Aspergillus niger A60: an evaluation of experimental methods.  

PubMed Central

The effects of elevated levels of dissolved carbon dioxide (dCO2), produced by gassing with CO2-enriched gas mixtures, upon an industrial strain of Aspergillus niger (strain A60) producing citrate and gluconate were quantitatively assessed. Particular attention was paid to the reliability and accuracy of the steam-sterilizable dCO2 probe, especially in the presence of high concentrations of potentially interfering acidic species. The response of the organism to elevated dCO2 levels was assessed by using both batch and chemostat cultures, and the sensitivity of the organism in different growth phases (lag, exponential, and stationary) was examined. Chemostat cultures showed markedly less inhibition (in terms of biomass and organic acid synthesis) than did batch cultures. Studies in batch culture indicated that lag-phase cultures were especially sensitive to elevated dCO2 levels. Overall, the results of this study indicate that previous experimental methods used to examine dCO2 effects in submerged cultures (continuous CO2-enriched gassing of batch cultures from time zero) have been inappropriate and have led to systematic overestimation of the inhibitory effects of dCO2 on mycelial organisms. PMID:9361401

McIntyre, M; McNeil, B

1997-01-01

345

Effect of Terminalia catappa Fruit Meal Fermented by Aspergillus niger as Replacement of Maize on Growth Performance, Nutrient Digestibility, and Serum Biochemical Profile of Broiler Chickens  

PubMed Central

A feeding experiment was conducted to investigate the effect of fermented Terminalia catappa fruit meal (FTCM) with Aspergillus niger as replacement for maize on broiler growth performance, nutrient digestibility, and serum biochemical constituents. Dietary maize was replaced by FTCM at 0, 20, 40, 60, or 80%. One hundred and eighty one-day-old Shaver broiler chicks were randomly allocated to the five dietary treatments, three replicate groups of twelve chicks each for a 42-day period. There was no significant difference (P > .05) in the feed intake, weight gain, and feed; gain ratio between the broilers fed on 40% FTCM diet and the control group. The apparent digestibilities of nitrogen, crude fibre, and fat decreased significantly in broilers fed higher levels (>40%) of FTCM replacement diets compared with the control or lower FTCM diets. Serum concentrations of total protein, albumin, and globulin were decreased (P < .05) on 80% FTCM fed broilers. Serum cholesterol, creatinine, and glucose were not significantly (P > .05) altered among treatments. The activities of aspartate and alanine aminotransferases and alkaline phosphatase were significantly (P < .05) increased with higher FTCM replacement. The results indicate that FTCM could replace up to 40% of dietary maize in the diets of broiler chickens without adverse effect on growth performance or serum constituents. PMID:21350670

Apata, David Friday

2011-01-01

346

Catalysis of Rice Straw Hydrolysis by the Combination of Immobilized Cellulase from Aspergillus niger on ?-Cyclodextrin-Fe3O4 Nanoparticles and Ionic Liquid  

PubMed Central

Cellulase from Aspergillus niger was immobilized onto ?-cyclodextrin-conjugated magnetic particles by silanization and reductive amidation. The immobilized cellulase gained supermagnetism due to the magnetic nanoparticles. Ninety percent of cellulase was immobilized, but the activity of immobilized cellulase decreased by 10%. In this study, ionic liquid (1-butyl-3-methylimidazolium chloride) was introduced into the hydrolytic process because the original reaction was a solid-solid reaction. The activity of immobilized cellulase was improved from 54.87 to 59.11?U?g immobilized cellulase?1 at an ionic liquid concentration of 200?mM. Using immobilized cellulase and ionic liquid in the hydrolysis of rice straw, the initial reaction rate was increased from 1.629 to 2.739?g?h?1?L?1. One of the advantages of immobilized cellulase is high reusability—it was usable for a total of 16 times in this study. Compared with free cellulase, magnetized cellulase can be recycled by magnetic field and the activity of immobilized cellulase was shown to remain at 85% of free cellulase without denaturation under a high concentration of glucose (15?g?L?1). Therefore, immobilized cellulase can hydrolyze rice straw continuously compared with free cellulase. The amount of harvested glucose can be up to twentyfold higher than that from the hydrolysis by free cellulase.

Huang, Po-Jung; Chang, Ken-Lin; Chen, Shui-Tein

2015-01-01

347

Total Phenolic Contents, Antioxidant Activities, and Lipid Fractions from Berry Pomaces Obtained by Solid-State Fermentation of Two Sambucus Species with Aspergillus niger.  

PubMed

The aim of this study was to investigate the effect of solid-state fermentation (SSF) by Aspergillus niger on phenolic contents and antioxidant activity in Sambucus nigra L. and Sambucus ebulus L. berry pomaces. The effect of fermentation time on the total fats and major lipid classes (neutral and polar) was also investigated. During the SSF, the extractable phenolics increased with 18.82% for S. ebulus L. and 11.11% for S. nigra L. The levels of antioxidant activity of methanolic extracts were also significantly enhanced. The HPLC-MS analysis indicated that the cyanidin 3-sambubioside-5-glucoside is the major phenolic compound in both fermented Sambucus fruit residues. In the early stages of fungal growth, the extracted oils (with TAGs as major lipid fraction) increased with 12% for S. nigra L. and 10.50% for S. ebulus L. The GC-MS analysis showed that the SSF resulted in a slight increase of the linoleic and oleic acids level. PMID:25787023

Dulf, Francisc Vasile; Vodnar, Dan Cristian; Dulf, Eva-Henrietta; To?a, Monica Ioana

2015-04-01

348

Production of cellulosic ethanol and enzyme from waste fiber sludge using SSF, recycling of hydrolytic enzymes and yeast, and recombinant cellulase-producing Aspergillus niger.  

PubMed

Bioethanol and enzymes were produced from fiber sludges through sequential microbial cultivations. After a first simultaneous saccharification and fermentation (SSF) with yeast, the bioethanol concentrations of sulfate and sulfite fiber sludges were 45.6 and 64.7 g/L, respectively. The second SSF, which included fresh fiber sludges and recycled yeast and enzymes from the first SSF, resulted in ethanol concentrations of 38.3 g/L for sulfate fiber sludge and 24.4 g/L for sulfite fiber sludge. Aspergillus niger carrying the endoglucanase-encoding Cel7B gene of Trichoderma reesei was grown in the spent fiber sludge hydrolysates. The cellulase activities obtained with spent hydrolysates of sulfate and sulfite fiber sludges were 2,700 and 2,900 nkat/mL, respectively. The high cellulase activities produced by using stillage and the significant ethanol concentrations produced in the second SSF suggest that onsite enzyme production and recycling of enzyme are realistic concepts that warrant further attention. PMID:24862324

Cavka, Adnan; Alriksson, Björn; Rose, Shaunita H; van Zyl, Willem H; Jönsson, Leif J

2014-08-01

349

Analysis of metal Bioleaching from thermal power plant fly ash by Aspergillus niger 34770 culture supernatant and reduction of phytotoxicity during the process.  

PubMed

Aspergillus niger culture supernatant is used for bioleaching process. Before starting bioleaching process, fly ash was washed with distilled water. This removed 100 % sodium, 47 % (±0.45) boron, 38.07 % (±0.12) calcium, 29.89 % (±0.78) magnesium, and 11.8 % (±0.05) potassium. The pH was reduced from 10.5 to 8.5 after water washing. During bioleaching process, around 100 % metal removal was achieved in 4 h for all metals except chromium 93 % (±1.18), nickel 83 % (±0.32), arsenic 78 % (±0.52), and lead 70 % (±0.20). The process parameters including temperature, shaking speed, and solid/liquid ratio were optimized for bioleaching process. Experiments were conducted to evaluate effect of fly ash on growth of mung bean (Vigna radiata). At 20 g/100 ml fly ash concentration no germination of V. radiata seeds was observed. With an increasing concentration of untreated fly ash, a gradual decrease in root/shoot length was observed. After bioleaching process 78 % (±0.19) germination of V. radiata was observed with 20 g/100 ml fly ash. This study will help to develop an efficient process to remove the toxic metals from fly ash. PMID:25349087

Jadhav, Umesh U; Hocheng, Hong

2015-01-01

350

Tamarind seed powder and palm kernel cake: two novel agro residues for the production of tannase under solid state fermentation by Aspergillus niger ATCC 16620.  

PubMed

Palm kernel cake (PKC), the residue obtained after extraction of palm oil from oil palm seeds and tamarind seed powder (TSP) obtained after removing the fruit pulp from tamarind fruit pod were tested for the production of tannase under solid-state fermentation (SSF) using Aspergillus niger ATCC 16620. The fungal strain was grown on the substrates without any pretreatment. In PKC medium, a maximum enzyme yield of 13.03 IU/g dry substrate (gds) was obtained when SSF was carried out at 30 degrees C, 53.5% initial substrate moisture, 33 x 10(9) spores/5 g substrate inoculum size and 5% tannic acid as additional carbon source after 96 h of fermentation. In TSP medium, maximum tannase yield of 6.44 IU/gds was obtained at 30 degrees C, 65.75% initial substrate moisture, 11 x 10(9) spores/5 g substrate inoculum, 1% glycerol as additional carbon source and 1% potassium nitrate as additional nitrogen source after 120 h of fermentation. Results from the study are promising for the economic utilization and value addition of these important agro residues, which are abundantly available in many tropical and subtropical countries. PMID:15734308

Sabu, A; Pandey, A; Daud, M Jaafar; Szakacs, G

2005-07-01

351

Identification of InuR, a new Zn(II)2Cys6 transcriptional activator involved in the regulation of inulinolytic genes in Aspergillus niger.  

PubMed

The expression of inulinolytic genes in Aspergillus niger is co-regulated and induced by inulin and sucrose. We have identified a positive acting transcription factor InuR, which is required for the induced expression of inulinolytic genes. InuR is a member of the fungal specific class of transcription factors of the Zn(II)2Cys6 type. Involvement of InuR in inulin and sucrose metabolism was suspected because of the clustering of inuR gene with sucB, which encodes an intracellular invertase with transfructosylation activity and a putative sugar transporter encoding gene (An15g00310). Deletion of the inuR gene resulted in a strain displaying a severe reduction in growth on inulin and sucrose medium. Northern analysis revealed that expression of inulinolytic and sucrolytic genes, e.g., inuE, inuA, sucA, as well as the putative sugar transporter gene (An15g00310) is dependent on InuR. Genome-wide expression analysis revealed, three additional putative sugar transporters encoding genes (An15g04060, An15g03940 and An17g01710), which were strongly induced by sucrose in an InuR dependent way. In silico analysis of the promoter sequences of strongly InuR regulated genes suggests that InuR might bind as dimer to two CGG triplets, which are separated by eight nucleotides. PMID:17917744

Yuan, Xiao-Lian; Roubos, Johannes A; van den Hondel, Cees A M J J; Ram, Arthur F J

2008-01-01

352

Effect of C/N Ratio and Media Optimization through Response Surface Methodology on Simultaneous Productions of Intra- and Extracellular Inulinase and Invertase from Aspergillus niger ATCC 20611  

PubMed Central

The study is to identify the extraction of intracellular inulinase (exo- and endoinulinase) and invertase as well as optimization medium composition for maximum productions of intra- and extracellular enzymes from Aspergillus niger ATCC 20611. From two different methods for extraction of intracellular enzymes, ultrasonic method was found more effective. Response surface methodology (RSM) with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. The effect of five main reaction parameters including sucrose, yeast extract, NaNO3, Zn+2, and Triton X-100 on the production of enzymes was analyzed. A modified quadratic model was fitted to the data with a coefficient of determination (R2) more than 0.90 for all responses. The intra-extracellular inulinase and invertase productions increased in the range from 16 to 8.4 times in the optimized medium (10% (w/v) sucrose, 2.5% (w/v) yeast extract, 2% (w/v) NaNO3, 1.5?mM (v/v) Zn+2, and 1% (v/v) Triton X-100) by RSM and from around 1.2 to 1.3 times greater than in the medium optimized by one-factor-at-a-time, respectively. The results of bioprocesses optimization can be useful in the scale-up fermentation and food industry. PMID:24151605

Dinarvand, Mojdeh; Rezaee, Malahat; Masomian, Malihe; Jazayeri, Seyed Davoud; Zareian, Mohsen; Abbasi, Sahar; Ariff, Arbakariya B.

2013-01-01

353

Sequence determination of a satellite RNA isolated from Aspergillus foetidus.  

PubMed

Aspergillus foetidus virus (AfV) has at least two distinct particle types, designated as AfV-fast (F) and AfV-slow (S). AfV-S includes AfV-S1, a victorivirus; AfV-S2, an unclassified satellite RNA; and AfV-S3, a previously uncharacterized dsRNA element. Here, we describe the complete sequence of AfV-S3, which is a short non-coding RNA with no known homologs. AfV-S3 is predicted to form an extended secondary structure, shares a 5' terminus with AfV-S2, and is a satellite RNA possibly dependent on both AfV-S1 and AfV-S2. This work concludes the sequencing of the A. foetidus virome. PMID:25613164

Shah, Unnati A; Kotta-Loizou, Ioly; Coutts, Robert H A

2015-03-01

354

Biodegradation of International Jet A-1 Aviation Fuel by Microorganisms Isolated from Aircraft Tank and Joint Hydrant Storage Systems  

Microsoft Academic Search

Microorganisms contaminating international Jet A-1 aircraft fuel and fuel preserved in Joint Hydrant Storage Tank (JHST) were\\u000a isolated, characterized and identified. The isolates were Bacillus subtillis, Bacillus megaterium, Flavobacterium oderatum, Sarcina flava, Micrococcus varians, Pseudomonas aeruginosa, Bacillus licheniformis, Bacillus cereus and Bacillus brevis. Others included Candida tropicalis, Candida albicans, Saccharomyces estuari, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Aspergillus flavus, Aspergillus niger, Aspergillus

A. Y. Itah; A. A. Brooks; B. O. Ogar; A. B. Okure

2009-01-01

355

Comparison of species composition and fumonisin production in Aspergillus section Nigri populations in maize kernels from USA and Italy.  

PubMed

Fumonisin contamination of maize is considered a serious problem in most maize-growing regions of the world, due to the widespread occurrence of these mycotoxins and their association with toxicosis in livestock and humans. Fumonisins are produced primarily by species of Fusarium that are common in maize grain, but also by some species of Aspergillus sect. Nigri, which can also occur on maize kernels as opportunistic pathogens. Understanding the origin of fumonisin contamination in maize is a key component in developing effective management strategies. Although some fungi in Aspergillus sect. Nigri are known to produce fumonisins, little is known about the species which are common in maize and whether they make a measurable contribution to fumonisin contamination of maize grain. In this work, we evaluated populations of Aspergillus sect. Nigri isolated from maize in USA and Italy, focusing on analysis of housekeeping genes, the fum8 gene and in vitro capability of producing fumonisins. DNA sequencing was used to identify Aspergillus strains belonging to sect. Nigri, in order to compare species composition between the two populations, which might influence specific mycotoxicological risks. Combined beta-tubulin/calmodulin sequences were used to genetically characterize 300 strains (199 from Italy and 101 from USA) which grouped into 4 clades: Aspergillus welwitschiae (syn. Aspergillus awamori, 14.7%), Aspergillus tubingensis (37.0%) and Aspergillus niger group 1 (6.7%) and group 2 (41.3%). Only one strain was identified as Aspergillus carbonarius. Species composition differed between the two populations; A. niger predominated among the USA isolates (69%), but comprised a smaller percentage (38%) of Italian isolates. Conversely, A. tubingensis and A. welwitschiae occurred at higher frequencies in the Italian population (42% and 20%, respectively) than in the USA population (27% and 5%). The evaluation of FB2 production on CY20S agar revealed 118 FB2 producing and 84 non-producing strains distributed among the clades: A. welwitschiae, A. niger group 1 and A. niger group 2, confirming the potential of Aspergillus sect. Nigri species to contribute to total fumonisin contamination of maize. A higher percentage of A. niger isolates (72.0%) produced FB2 compared to A. welwitschiae (36.6%). The percentage of FB2-producing A. niger strains was similar in the USA and Italian populations; however, the predominance of A. niger in the USA population suggests a higher potential for fumonisin production. Some strains with fum8 present in the genome did not produce FB2in vitro, confirming the ineffectiveness of fum8 presence as a predictor of FB2 production. PMID:25087207

Susca, Antonia; Moretti, Antonio; Stea, Gaetano; Villani, Alessandra; Haidukowski, Miriam; Logrieco, Antonio; Munkvold, Gary

2014-10-01

356

Impact of alg3 gene deletion on growth, development, pigment production, protein secretion, and functions of recombinant Trichoderma reesei cellobiohydrolases in Aspergillus niger.  

PubMed

Dolichyl-P-Man:Man(5)GlcNAc(2)-PP-dolichyl ?-1,3-mannosyltransferase (also known as "asparagine-linked glycosylation 3", or ALG3) is involved in early N-linked glycan synthesis and thus is essential for formation of N-linked protein glycosylation. In this study, we examined the effects of alg3 gene deletion (alg3?) on growth, development, pigment production, protein secretion and recombinant Trichoderma reesei cellobiohydrolase (rCel7A) expressed in Aspergillus niger. The alg3? delayed spore germination in liquid cultures of complete medium (CM), potato dextrose (PD), minimal medium (MM) and CM with addition of cAMP (CM+cAMP), and resulted in significant reduction of hyphal growth on CM, potato dextrose agar (PDA), and CM+cAMP and spore production on CM. The alg3? also led to a significant accumulation of red pigment on both liquid and solid CM cultures. The relative abundances of 54 of the total 215 proteins identified in the secretome were significantly altered as a result of alg3?, 63% of which were secreted at higher levels in alg3? strain than the parent. The rCel7A expressed in the alg3? mutant was smaller in size than that expressed in both wild-type and parental strains, but still larger than T. reesei Cel7A. The circular dichroism (CD)-melt scans indicated that change in glycosylation of rCel7A does not appear to impact the secondary structure or folding. Enzyme assays of Cel7A and rCel7A on nanocrystalline cellulose and bleached kraft pulp demonstrated that the rCel7As have improved activities on hydrolyzing the nanocrystalline cellulose. Overall, the results suggest that alg3 is critical for growth, sporulation, pigment production, and protein secretion in A. niger, and demonstrate the feasibility of this alternative approach to evaluate the roles of N-linked glycosylation in glycoprotein secretion and function. PMID:24076077

Dai, Ziyu; Aryal, Uma K; Shukla, Anil; Qian, Wei-Jun; Smith, Richard D; Magnuson, Jon K; Adney, William S; Beckham, Gregg T; Brunecky, Roman; Himmel, Michael E; Decker, Stephen R; Ju, Xiaohui; Zhang, Xiao; Baker, Scott E

2013-12-01

357

A novel glucose dehydrogenase from the white-rot fungus Pycnoporus cinnabarinus: production in Aspergillus niger and physicochemical characterization of the recombinant enzyme.  

PubMed

Data on glucose dehydrogenases (GDHs) are scarce and availability of these enzymes for application purposes is limited. This paper describes a new GDH from the fungus Pycnoporus cinnabarinus CIRM BRFM 137 that is the first reported GDH from a white-rot fungus belonging to the Basidiomycota. The enzyme was recombinantly produced in Aspergillus niger, a well-known fungal host producing an array of homologous or heterologous enzymes for industrial applications. The full-length gene that encodes GDH from P. cinnabarinus (PcGDH) consists of 2,425 bp and codes for a deduced protein of 620 amino acids with a calculated molecular mass of 62.5 kDa. The corresponding complementary DNA was cloned and placed under the control of the strong and constitutive glyceraldehyde-3-phosphate dehydrogenase promoter. The signal peptide of the glucoamylase prepro sequence of A. niger was used to target PcGDH secretion into the culture medium, achieving a yield of 640 mg L(-1), which is tenfold higher than any other reported value. The recombinant PcGDH was purified twofold to homogeneity in a one-step procedure with a 41 % recovery using a Ni Sepharose column. The identity of the recombinant protein was further confirmed by immunodetection using western blot analysis and N-terminal sequencing. The molecular mass of the native PcGDH was 130 kDa, suggesting a homodimeric form. Optimal pH and temperature were found to be similar (5.5 and 60 °C, respectively) to those determined for the previously characterized GDH, i.e., from Glomerella cingulata. However PcGDH exhibits a lower catalytic efficiency of 67 M(-1) s(-1) toward glucose. This substrate is by far the preferred substrate, which constitutes an advantage over other sugar oxidases in the case of blood glucose monitoring. The substrate-binding domain of PcGDH turns out to be conserved as compared to other glucose-methanol-choline (GMCs) oxidoreductases. In addition, the ability of PcGDH to reduce oxidized quinones or radical intermediates was clearly demonstrated, which raises prospects for applying this enzyme to detoxify toxic compounds formed during the degradation of lignin. PMID:24965558

Piumi, François; Levasseur, Anthony; Navarro, David; Zhou, Simeng; Mathieu, Yann; Ropartz, David; Ludwig, Roland; Faulds, Craig B; Record, Eric

2014-12-01

358

In vitro activity of Isavuconazole against Aspergillus species and zygomycetes according to the methodology of the European Committee on Antimicrobial Susceptibility Testing.  

PubMed

We evaluated the MICs of isavuconazole (ISAV) against 96 isolates of Aspergillus species and 36 zygomycetes according to the methodology of the European Committee on Antimicrobial Susceptibility Testing. In addition, the in vitro activity was obtained for hyphal inocula. ISAV exhibited good antifungal activity against the tested isolates with the exception of Aspergillus niger and Mucorales. The in vitro activity of ISAV was comparable to that of voriconazole aside from Mucorales. PMID:19164153

Perkhofer, Susanne; Lechner, Veronika; Lass-Flörl, Cornelia

2009-04-01

359

In vitro susceptibility of 188 clinical and environmental isolates of Aspergillus flavus for the new triazole isavuconazole and seven other antifungal drugs  

Microsoft Academic Search

Recently isavuconazole, an experimental triazole agent, was found to be active against Aspergillus species. As Aspergillus flavus is the second-most common Aspergillus species isolated from human infection and the fungus has not been widely tested against the drug, we studied a large collection of clinical (n = 178) and environmental (n = 10) strains of A. flavus against isavuconazole and

M. R. Shivaprakash; E. Geertsen; A. Chakrabarti; J. W. Mouton; J. F. G. M. Meis

2011-01-01

360

Cloning, heterologous expression, purification and characterization of M12 mutant of Aspergillus niger glucose oxidase in yeast Pichia pastoris KM71H.  

PubMed

Aspergillus niger glucose oxidase (GOx) genes for wild-type (GenBank accession no. X16061, swiss-Prot; P13006) and M12 mutant (N2Y, K13E, T30 V, I94 V, K152R) were cloned into pPICZ?A vector for expression in Pichia pastoris KM71H strain. The highest expression level of 17.5 U/mL of fermentation media was obtained in 0.5 % (v/v) methanol after 9 days of fermentation. The recombinant GOx was purified by cross-flow ultrafiltration using membranes of 30 kDa molecular cutoff and DEAE ion-exchange chromatography at pH 6.0. Purified wt GOx had k cat of 189.4 s?¹ and K(m) of 28.26 mM while M12 GOx had k cat of 352.0 s?¹ and K m of 13.33 mM for glucose at pH 5.5. Specificity constants k(cat)/K(m) of wt (6.70 mM?¹ s?¹) and M12 GOx (26.7 mM?¹ s?¹) expressed in P. pastoris KM71H were around three times higher than for the same enzymes previously expressed in Saccharomyces cerevisiae InvSc1 strain. The pH optimum and sugar specificity of M12 mutant of GOx remained similar to the wild-type form of the enzyme, while thermostability was slightly decreased. M12 GOx expressed in P. pastoris showed three times higher activity compared to the wt GOx toward redox mediators like N,N-dimethyl-nitroso-aniline used for glucose strips manufacturing. M12 mutant of GOx produced in P. pastoris KM71H could be useful for manufacturing of glucose biosensors and biofuel cells. PMID:24122283

Kova?evi?, Gordana; Blaži?, Marija; Dragani?, Bojana; Ostafe, Raluca; Gavrovi?-Jankulovi?, Marija; Fischer, Rainer; Prodanovi?, Radivoje

2014-04-01

361

Shifting the pH Profile of Aspergillus niger PhyA Phytase To Match the Stomach pH Enhances Its Effectiveness as an Animal Feed Additive  

PubMed Central

Environmental pollution by phosphorus from animal waste is a major problem in agriculture because simple-stomached animals, such as swine, poultry, and fish, cannot digest phosphorus (as phytate) present in plant feeds. To alleviate this problem, a phytase from Aspergillus niger PhyA is widely used as a feed additive to hydrolyze phytate-phosphorus. However, it has the lowest relative activity at the pH of the stomach (3.5), where the hydrolysis occurs. Our objective was to shift the pH optima of PhyA to match the stomach condition by substituting amino acids in the substrate-binding site with different charges and polarities. Based on the crystal structure of PhyA, we prepared 21 single or multiple mutants at Q50, K91, K94, E228, D262, K300, and K301 and expressed them in Pichia pastoris yeast. The wild-type (WT) PhyA showed the unique bihump, two-pH-optima profile, whereas 17 mutants lost one pH optimum or shifted the pH optimum from pH 5.5 to the more acidic side. The mutant E228K exhibited the best overall changes, with a shift of pH optimum to 3.8 and 266% greater (P < 0.05) hydrolysis of soy phytate at pH 3.5 than the WT enzyme. The improved efficacy of the enzyme was confirmed in an animal feed trial and was characterized by biochemical analysis of the purified mutant enzymes. In conclusion, it is feasible to improve the function of PhyA phytase under stomach pH conditions by rational protein engineering. PMID:16751556

Kim, Taewan; Mullaney, Edward J.; Porres, Jesus M.; Roneker, Karl R.; Crowe, Sarah; Rice, Sarah; Ko, Taegu; Ullah, Abul H. J.; Daly, Catherine B.; Welch, Ross; Lei, Xin Gen

2006-01-01

362

Cloning, expression in Pichia pastoris, and characterization of a thermostable GH5 mannan endo-1,4-?-mannosidase from Aspergillus niger BK01  

PubMed Central

Background Mannans are key components of lignocellulose present in the hemicellulosic fraction of plant primary cell walls. Mannan endo-1,4-?-mannosidases (1,4-?-D-mannanases) catalyze the random hydrolysis of ?-1,4-mannosidic linkages in the main chain of ?-mannans. Biodegradation of ?-mannans by the action of thermostable mannan endo-1,4-?-mannosidase offers significant technical advantages in biotechnological industrial applications, i.e. delignification of kraft pulps or the pretreatment of lignocellulosic biomass rich in mannan for the production of second generation biofuels, as well as for applications in oil and gas well stimulation, extraction of vegetable oils and coffee beans, and the production of value-added products such as prebiotic manno-oligosaccharides (MOS). Results A gene encoding mannan endo-1,4-?-mannosidase or 1,4-?-D-mannan mannanohydrolase (E.C. 3.2.1.78), commonly termed ?-mannanase, from Aspergillus niger BK01, which belongs to glycosyl hydrolase family 5 (GH5), was cloned and successfully expressed heterologously (up to 243 ?g of active recombinant protein per mL) in Pichia pastoris. The enzyme was secreted by P. pastoris and could be collected from the culture supernatant. The purified enzyme appeared glycosylated as a single band on SDS-PAGE with a molecular mass of approximately 53 kDa. The recombinant ?-mannanase is highly thermostable with a half-life time of approximately 56 h at 70°C and pH 4.0. The optimal temperature (10-min assay) and pH value for activity are 80°C and pH 4.5, respectively. The enzyme is not only active towards structurally different mannans but also exhibits low activity towards birchwood xylan. Apparent Km values of the enzyme for konjac glucomannan (low viscosity), locust bean gum galactomannan, carob galactomannan (low viscosity), and 1,4-?-D-mannan (from carob) are 0.6 mg mL-1, 2.0 mg mL-1, 2.2 mg mL-1 and 1.5 mg mL-1, respectively, while the kcat values for these substrates are 215 s-1, 330 s-1, 292 s-1 and 148 s-1, respectively. Judged from the specificity constants kcat/Km, glucomannan is the preferred substrate of the A. niger ? -mannanase. Analysis by thin layer chromatography showed that the main product from enzymatic hydrolysis of locust bean gum is mannobiose, with only low amounts of mannotriose and higher manno-oligosaccharides formed. Conclusion This study is the first report on the cloning and expression of a thermostable mannan endo-1,4-?-mannosidase from A. niger in Pichia pastoris. The efficient expression and ease of purification will significantly decrease the production costs of this enzyme. Taking advantage of its acidic pH optimum and high thermostability, this recombinant ?-mannanase will be valuable in various biotechnological applications. PMID:19912637

2009-01-01

363

Molecular characterization of aflatoxigenic and non-aflatoxigenic Aspergillus flavus isolates collected from corn grains.  

PubMed

Twelve species from six fungal genera were found to be associated with corn (Zea mays L.) grain samples collected from three main regions of Saudi Arabia. The average frequencies of the most common genera were Aspergillus (11.4%), Fusarium (9.5%), Penicillium (5.1%), and Alternaria (5.8%). Fifteen isolates of Aspergillus flavus were screened by HPLC for their ability to produce aflatoxins (AF). The percentage of aflatoxigenic A. flavus isolates was 53%. Eight isolates produced AF, at concentrations ranging 0.7-2.9 ppb. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular markers were used to genetically characterize isolates of A. flavus and to discriminate between the aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, which was useful for genetic characterization. The clustering in the RAPD and ISSR dendrograms obtained was unrelated to geographic origin. The RAPD and ISSR markers could not discriminate between aflatoxigenic and non-aflatoxigenic isolates, but the ISSR primers were somewhat better. PMID:25501147

Mahmoud, M A; Ali, H M; El-Aziz, A R M; Al-Othman, M R; Al-Wadai, A S

2014-01-01

364

Aerial Prevalence of Aspergillus calidoustus Isolates in and around a Tertiary Care Hospital in Kuwait and Assessment of Their Pathogenicity  

PubMed Central

Seven Aspergillus calidoustus isolates from 486 Aspergillus spp. isolates (1.4% overall prevalence) from outdoor/indoor air samples and one isolate from the bronchoalveolar lavage fluid of a patient with pneumonia were obtained. These 8 isolates exhibited reduced susceptibility to triazoles. Preliminary pathogenicity data from BALB/c mice suggest that A. calidoustus can persist in tissues for long periods without causing mortality. Further studies using graded doses of inoculum and immunosuppression models are warranted to gain an understanding of the factors associated with its pathogenicity and virulence. PMID:24920775

Ahmad, Suhail; Joseph, Leena

2014-01-01

365

Antifungal susceptibility of Aspergillus fumigatus clinical isolates collected from various areas in Japan.  

PubMed

Azole resistance among clinical isolates of Aspergillus fumigatus is becoming a serious problem in Europe, but the status in Japan is not yet known in detail. The aim of this study was to determine the present status of azole resistance in A. fumigatus in Japan. We employed 171 clinical isolates of A. fumigatus sensu stricto collected from 1987 to 2008 at the Medical Mycology Research Center, Chiba University, Japan for azole resistance determination. Identification of all isolates were re-examined both from the aspect of morphology and molecular phylogeny. The antifungal susceptibility of these isolates was tested based on the CLSI M38-A2 broth microdilution method. In our collection, only 1 (0.6%) and 2 isolates (1.2%) showed elevated MIC to voriconazole and itraconazole, respectively. Our study disclosed that the frequency of azole resistance in A. fumigatus still remains low in this collection. PMID:24751235

Kikuchi, Kazuyo; Watanabe, Akira; Ito, Junko; Oku, Yukio; Wuren, Tuya; Taguchi, Hideaki; Yarita, Kyoko; Muraosa, Yasunori; Yahiro, Maki; Yaguchi, Takashi; Kamei, Katsuhiko

2014-05-01

366

Molecular identification and antifungal susceptibility profile of Aspergillus flavus isolates recovered from clinical specimens in Kuwait  

PubMed Central

Background Within the genus Aspergillus, A. flavus is the second most important species of clinical significance. It is predominantly associated with infections involving sinuses, eye and skin, mostly in geographic regions with hot and arid climate, including the Middle East. Recent reports on emergence of resistance to triazoles among Aspergillus spp. is a cause of concern for treatment of patients with invasive aspergillosis. In this study we present data on genetic characterization and antifungal susceptibility profile of clinical and environmental isolates of A. flavus. Methods Ninety-nine Aspergillus section Flavi isolates, originating from clinical (n=92) and environmental (n=7) sources, initially identified by morphological characteristics, were analyzed by partial sequencing of ?-tubulin and calmodulin gene fragments and their susceptibilities to six antifungal agents was determined by Etest on RPMI1640 and Muller-Hinton agar media. Etest minimum inhibitory concentrations (MICs) of amphotericin B and voriconazole were also compared with zone of inhibition diameters obtained by disc diffusion test on RPMI agar medium. Results The identity of all clinical and environmental isolates was confirmed as A. flavus species by combined analysis of ?-tubulin and calmodulin genes. The mean MIC90 (?g/ml) values on RPMI medium for amphotericin B, voriconazole, posaconazole, anidulafungin, micafungin and caspofungin were 3, 0.25, 0.25, 0.002, 0.002 and 0.032, respectively. No environmental isolate exhibited MIC value of >2 ?g/ml for amphotericin B. For clinical isolates, the zone of inhibition diameters for amphotericin B and voriconazole ranged from 7–16 mm and 24–34 mm, respectively. Linear regression analysis between Etest MIC values and disk diffusion diameters revealed a significant inverse correlation with amphotericin B (p <0.001) and voriconazole (p<0.003). Conclusions The ?-tubulin and calmodulin gene sequences confirmed that all 92 clinical isolates identified phenotypically belonged to A. flavus taxon, thus suggesting that the other species within Aspergillus section Flavi are of little clinical significance. Triazoles and echinocandins showed very good in vitro activity against the A. flavus, however, 10% clinical isolates showed MICs of >2 ?g/ml for amphotericin B. PMID:23496810

2013-01-01

367

Aspergillus 6V4, a Strain Isolated from Manipueira, Produces High Amylases Levels by Using Wheat Bran as a Substrate  

PubMed Central

The aim of this study was screening fungi strains, isolated from manipueira (a liquid subproduct obtained from the flour production of Manihot esculenta), for amylases production and investigating production of these enzymes by the strain Aspergillus 6V4. The fungi isolated from manipueira belonged to Ascomycota phylum. The strain Aspergillus 6V4 was the best amylase producer in the screening assay of starch hydrolysis in petri dishes (ASHPD) and in the assay in submerged fermentation (ASbF). The strain Aspergillus 6V4 produced high amylase levels (335?UI/L) using wheat bran infusion as the exclusive substrate and the supplementation of this substrate with peptone decreased the production of this enzyme. The moisture content of 70% was the best condition for the production of Aspergillus 6V4 amylases (385?IU/g) in solid state fermentation (SSF). PMID:24724017

Celestino, Jessyca dos Reis; Duarte, Ana Caroline; Silva, Cláudia Maria de Melo; Sena, Hellen Holanda; Ferreira, Maria do Perpétuo Socorro Borges Carriço; Mallmann, Neila Hiraishi; Lima, Natacha Pinheiro Costa; Tavares, Chanderlei de Castro; de Souza, Rodrigo Otávio Silva; Souza, Érica Simplício; Souza, João Vicente Braga

2014-01-01

368

New and revisited species in Aspergillus section Nigri.  

PubMed

Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indologenus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them based on either ?-tubulin or calmodulin sequence data. Aspergillus eucalypticola produced pyranonigrin A, funalenone, aurasperone B and other naphtho-?-pyrones. Aspergillus neoniger is also a biseriate species isolated from desert sand in Namibia, and mangrove water in Venezuela, which produces aurasperone B and pyranonigrin A. Aspergillus fijiensis is a uniseriate species related to A. aculeatinus, and was isolated from soil in Fiji, and from Lactuca sativa in Indonesia. This species is able to grow at 37 °C, and produces asperparalines and okaramins. Aspergillus indologenus was isolated from soil, India. This species also belongs to the uniseriate group of black aspergilli, and was found to be related to, but clearly distinguishable from A. uvarum based on ?-tubulin, calmodulin and ITS sequence data. Aspergillus indologenus produced the insecticidal compounds okaramins A, B, H, and two types of indol-alkaloids which have not been structure elucidated. Two other species, A. violaceofuscus and A. acidus, are revalidated based on molecular and extrolite data. Aspergillus violaceofuscus was found to be related to A. japonicus, and produced some of the same interesting indol-alkaloids as A. indologenus, and also produced several families of partially characterised extrolites that were also found in A. heteromorphus. Aspergillus acidus (previously known as A. foetidus var. pallidus and A. foetidus var. acidus) is also a valid species, while A. foetidus is a synonym of A. niger based on molecular and physiological data. Two other species described previously, A. coreanus and A. lacticoffeatus, were found to be colour mutants of A. acidus and A. niger, respectively. Methods which could be used to distinguish the two closely related and economically important species A. niger and A. awamori are also detailed. Although these species differ in their occurrence and several physiological means (elastase activities, abilities to utilise 2-deoxy-D-glucose as sole carbon source), our data indicate that only molecular approaches including sequence analysis of calmodulin or ?-tubulin genes, AFLP analysis, UP-PCR analysis or mtDNA RFLP analysis can be used reliably to distinguish these sibling species. Aspergillus section Nigri now includes 26 taxa. PMID:21892239

Varga, J; Frisvad, J C; Kocsubé, S; Brankovics, B; Tóth, B; Szigeti, G; Samson, R A

2011-06-30

369

Isolate-Dependent Growth, Virulence, and Cell Wall Composition in the Human Pathogen Aspergillus fumigatus  

PubMed Central

The ubiquitous fungal pathogen Aspergillus fumigatus is a mediator of allergic sensitization and invasive disease in susceptible individuals. The significant genetic and phenotypic variability between and among clinical and environmental isolates are important considerations in host-pathogen studies of A. fumigatus-mediated disease. We observed decreased radial growth, rate of germination, and ability to establish colony growth in a single environmental isolate of A. fumigatus, Af5517, when compared to other clinical and environmental isolates. Af5517 also exhibited increased hyphal diameter and cell wall ?-glucan and chitin content, with chitin most significantly increased. Morbidity, mortality, lung fungal burden, and tissue pathology were decreased in neutropenic Af5517-infected mice when compared to the clinical isolate Af293. Our results support previous findings that suggest a correlation between in vitro growth rates and in vivo virulence, and we propose that changes in cell wall composition may contribute to this phenotype. PMID:24945802

Amarsaikhan, Nansalmaa; O’Dea, Evan M.; Tsoggerel, Angar; Owegi, Henry; Gillenwater, Jordan; Templeton, Steven P.

2014-01-01

370

Isolation and chemical characterization of naphthoquinone metabolites of Aspergillus parvulus Smith  

SciTech Connect

Although several benzoquinone and anthraquinone compounds have been isolated from Aspergillus species, only two naphthoquinone monomers have been reported thus far. Aspergillus parvulus Smith (ATCC number16911) was first investigated chemically in 1974, and five naphthalenones, along with one naphthoquinone, were isolated and characterized. Based on biosynthetic considerations, it was thought that A. parvulus might be capable of producing additional naphthoquinones under suitable conditions. It was decided to undertake a further investigation of A. parvulus. Thus, three novel naphthoquinones, compounds A, B, and C, were isolated from A. parvulus cultures grown in an acidic medium of glucose and phytone peptone. The structures of these compounds were deduced largely by the comparison of the effects of acetylation on the /sup 1/H-NMR and /sup 13/C-NMR spectra of the parent compounds and their four derivatives. An unusual mass fragmentation pattern which was previously thought to be unfavorable was discovered, and the other fragmentation patterns of the parent compounds, as well as their derivatives, were proposed. This investigation appears to be the third reported isolation of 2,5,7-tri-hydroxy-1,4-naphthoquinone derivatives from nature and the first reported from A. parvulus.

Wang, C.C.P.

1984-01-01

371

The influence of Aspergillus niger transcription factors AraR and XlnR in the gene expression during growth in D-xylose, L-arabinose and steam-exploded sugarcane bagasse.  

PubMed

The interest in the conversion of plant biomass to renewable fuels such as bioethanol has led to an increased investigation into the processes regulating biomass saccharification. The filamentous fungus Aspergillus niger is an important microorganism capable of producing a wide variety of plant biomass degrading enzymes. In A. niger the transcriptional activator XlnR and its close homolog, AraR, controls the main (hemi-)cellulolytic system responsible for plant polysaccharide degradation. Sugarcane is used worldwide as a feedstock for sugar and ethanol production, while the lignocellulosic residual bagasse can be used in different industrial applications, including ethanol production. The use of pentose sugars from hemicelluloses represents an opportunity to further increase production efficiencies. In the present study, we describe a global gene expression analysis of A. niger XlnR- and AraR-deficient mutant strains, grown on a D-xylose/L-arabinose monosaccharide mixture and steam-exploded sugarcane bagasse. Different gene sets of CAZy enzymes and sugar transporters were shown to be individually or dually regulated by XlnR and AraR, with XlnR appearing to be the major regulator on complex polysaccharides. Our study contributes to understanding of the complex regulatory mechanisms responsible for plant polysaccharide-degrading gene expression, and opens new possibilities for the engineering of fungi able to produce more efficient enzymatic cocktails to be used in biofuel production. PMID:23892063

de Souza, Wagner Rodrigo; Maitan-Alfenas, Gabriela Piccolo; de Gouvêa, Paula Fagundes; Brown, Neil Andrew; Savoldi, Marcela; Battaglia, Evy; Goldman, Maria Helena S; de Vries, Ronald P; Goldman, Gustavo Henrique

2013-11-01

372

Whole-genome comparison of Aspergillus fumigatus strains serially isolated from patients with aspergillosis.  

PubMed

The emergence of azole-resistant strains of Aspergillus fumigatus during treatment for aspergillosis occurs by a mutation selection process. Understanding how antifungal resistance mechanisms evolve in the host environment during infection is of great clinical importance and biological interest. Here, we used next-generation sequencing (NGS) to identify mutations that arose during infection by A. fumigatus strains sequentially isolated from two patients, one with invasive pulmonary aspergillosis (IPA) (five isolations) and the other with aspergilloma (three isolations). The serial isolates had identical microsatellite types, but their growth rates and conidia production levels were dissimilar. A whole-genome comparison showed that three of the five isolates from the IPA patient carried a mutation, while 22 mutations, including six nonsynonymous ones, were found among three isolates from the aspergilloma patient. One aspergilloma isolate carried the cyp51A mutation P216L, which is reported to confer azole resistance, and it displayed an MIC indicating resistance to itraconazole. This isolate harbored five other nonsynonymous mutations, some of which were found in the afyap1 and aldA genes. We further identified a large deletion in the aspergilloma isolate in a region containing 11 genes. This finding suggested the possibility that genomic deletions can occur during chronic infection with A. fumigatus. Overall, our results revealed dynamic alterations that occur in the A. fumigatus genome within its host during infection and treatment. PMID:25232160

Hagiwara, Daisuke; Takahashi, Hiroki; Watanabe, Akira; Takahashi-Nakaguchi, Azusa; Kawamoto, Susumu; Kamei, Katsuhiko; Gonoi, Tohru

2014-12-01

373

Amylose-like polysaccharide accumulation and hyphal cell-surface structure in relation to citric acid production by Aspergillus niger in shake culture.  

PubMed

When 120 mg glucose/ml was used as a carbon source, in shake culture Aspergillus niger Yang no. 2 maximally produced only 15.4 mg citric acid/ml but accumulated 3.0 mg extracellular polysaccharide/ml. The polysaccharide secreted by mycelia of Yang no. 2 in shake culture was confirmed to be an amylose-like alpha-1,4-glucan by hydrolysis analysis with acid, amylase and glucoamylase. However, in static cultures, such as semisolid and surface cultures free from physical stresses caused by shaking damage, Yang no. 2 produced more citric acid but did not accumulate the polysaccharide. With cultivation time in shake culture, the amount of extracellular polysaccharide and the viscosity of the culture broth increased. The increase of shaking speed caused a remarkable increase in the accumulation of extracellular polysaccharide, e.g. 11.2 mg extracellular polysaccharide/ml was accumulated in the medium at a shaking speed of 200 rpm. The addition of 2.0 mg carboxymethylcellulose (CMC)/ml as a viscous additive to the medium reduced drastically the amount of extracellular polysaccharide accumulated to 1.5 mg/ml, but increased the citric acid produced to 52.0 mg/ml. However, intracellular polysaccharide accumulation kept up a steady rate of 0.26 microgram/mg dried mycelium through the entire period of cultivation. The addition of 3.0 mg polysaccharide/ml purified from the culture broth to the medium at the start of a culture resulted in a decrease of extracellular polysaccharide accumulation but an increase of citric acid accumulation. From electronmicroscopic observation, cell surfaces of hyphae cultivated with CMC were smooth, while hyphae cultivated without CMC had fibrous and granular polysaccharide on the cell surface. These results suggested that Yang no. 2 secreted the polysaccharide on the cell surface as a viscous substance and/or a shock absorber to protect itself from physical stresses caused by shaking damage in shake culture. PMID:10531655

Kirimura, K; Yusa, S; Rugsaseel, S; Nakagawa, H; Osumi, M; Usami, S

1999-09-01

374

Marine isolates of Aspergillus flavus: denizens of the deep or lost at sea?  

PubMed Central

Most fungal species from marine environments also live on land. It is not clear whether these fungi reach the sea from terrestrial sources as spores or other propagules, or if there are separate ecotypes that live and reproduce in the sea. The emergence of marine diseases has created an urgency to understand the distribution of these fungi. Aspergillus flavus is ubiquitous in both terrestrial and marine environments. This species is an opportunistic pathogen in many hosts, making it a good model to study the relationship between genetic diversity and specificity of marine fungi. In this study, an intraspecific phylogeny of A. flavus isolates based on Amplified Fragment Length Polymorphisms (AFLPs) was used to determine if terrestrial and marine isolates form discrete populations, and to determine if phylogeny predicts substratum specificity. Results suggest lack of population structure in A. flavus. All isolates may compose a single population, with no clade particular to marine environments. PMID:21076639

ZULUAGA-MONTERO, Anabella; RAMÍREZ-CAMEJO, Luis; RAUSCHER, Jason; BAYMAN, Paul

2010-01-01

375

Mutations in the cyp51A gene and susceptibility to itraconazole in Aspergillus fumigatus serially isolated from a patient with lung aspergilloma  

Microsoft Academic Search

Objectives: To monitor changes in itraconazole susceptibility of isolates from a patient undergoing treatment for pulmonary Aspergillus infection and relate these changes to genotypic\\/phenotypic alterations. Methods: Six Aspergillus fumigatus isolates were serially recovered from the patient. Itraconazole MICs were determined by Etest and NCCLS methodology. Growth characteristics and phenotype were monitored. Molecular analysis included random amplified polymorphic DNA (RAPD) assay

Jian Chen; Houmin Li; Ruoyu Li; Dingfang Bu; Zhe Wan

2005-01-01

376

Isolation of notoamide S and enantiomeric 6-epi-stephacidin A from the fungus Aspergillus amoenus: biogenetic implications.  

PubMed

Notoamide S has been hypothesized to be a key biosynthetic intermediate for characteristic metabolites stephacidin A, notoamide B, and versicolamide B in Aspergillus sp. but has not yet been isolated. The isolation of notoamide S and an enantiomeric mixture of 6-epi-stephacidin A enriched with the (-)-isomer from Aspergillus amoenus is reported. The presence of (+)-versicolamide B suggests that the fungus possesses only the oxidase, which converts (+)-6-epi-stephacidin A into (+)-Versicolamide B, but not for (-)-6-epi-Stephacidin A. PMID:25615822

Kato, Hikaru; Nakahara, Takashi; Sugimoto, Kayo; Matsuo, Kanae; Kagiyama, Ippei; Frisvad, Jens C; Sherman, David H; Williams, Robert M; Tsukamoto, Sachiko

2015-02-01

377

Genetic structure of Aspergillus flavus populations in human and avian isolates.  

PubMed

Aspergillus flavus is the second leading cause of allergic, invasive, and colonizing fungal diseases in humans, and also the second most frequent organism associated with avian infections. Currently, it is not known whether there is a link between the environmental isolates and/or human isolates of A. flavus and those responsible for aspergillosis in birds. Microsatellite typing was used to analyze 29 A. flavus clinical and environmental avian isolates and 63 human clinical isolates collected from patients with a variety of aspergillosis diseases. The combination of all six markers yielded 77 different genotypes with a 0.98 D value. A. flavus genotypes obtained from avian isolates were compared with those obtained from human clinical and environmental samples. The standardized indices of association I (A) and rBarD were significantly different from zero (p?isolates. The human environmental population was significantly differentiated from environmental and clinical avian populations (F (st)?>?0.25). The avian clinical subpopulation exchanged few strains with the environmental human (N (m)?=?7.24) and avian (N (m)?=?6.60) populations. The minimum spanning tree analysis identified three A. flavus genotype clusters that were highly structured according to the isolation source (p?

Hadrich, I; Amouri, I; Neji, S; Mahfoud, N; Ranque, S; Makni, F; Ayadi, A

2013-02-01

378

Sequence determination of a quadripartite dsRNA virus isolated from Aspergillus foetidus.  

PubMed

Virus infection of Aspergillus foetidus was documented over 40 years ago and was one of the first mycovirus infections described in a filamentous fungus. The virus, named Aspergillus foetidus virus (AfV), contains at least two types of icosahedral particles, called AfV-fast (-F) and AfV-slow (-S) virions, based on their relative electrophoretic mobilities. Here, we report the complete nucleotide sequence of the AfV-F genome isolated from virions purified from the prototype isolate of the fungus. The AfV-F double-stranded (ds) RNA genome is tetra-segmented, and the plus strands of each of the four segments, but not the minus strands, are polyadenylated. The organisation and sequences of the four AfV-F dsRNAs are similar to those described for Alternaria alternata virus 1, which we propose is a member of an emerging mycovirus genus ("Alternavirus") and family ("Alternaviridae"), which also includes AfV-F. PMID:22760661

Kozlakidis, Zisis; Herrero, Noemi; Ozkan, Selin; Kanhayuwa, Lakkhana; Jamal, Atif; Bhatti, Muhammad F; Coutts, Robert H A

2013-01-01

379

Biological control of Botrytis, Aspergillus and Rhizopus rots on table and wine grapes in Israel  

Microsoft Academic Search

One hundred and twenty-nine strains of epiphytic micro-organisms, isolated from table and wine grapes in Israel, were screened for antagonistic activity against Botrytis cinerea on table grapes. Two isolates (Candida guilliermondii, strain A42 and Acremonium cephalosporium, strain B11) were further evaluated for the control of decay in grapes caused by Aspergillus niger and Rhizopus stolonifer. Decay incidence caused by Botrytis

Tirtza Zahavi; Lea Cohen; Batia Weiss; Leonardo Schena; Avinoam Daus; Tania Kaplunov; Johanan Zutkhi; Ruth Ben-Arie; Samir Droby

2000-01-01

380

Identification of fungi of the genus Aspergillus section nigri using polyphasic taxonomy  

PubMed Central

In spite of the taxonomy of the Aspergillus species of the Nigri Section being regarded as troublesome, a number of methods have been proposed to aid in the classification of this Section. This work aimed to distinguish Aspergillus species of the Nigri Section from foods, grains and caves on the basis in Polyphasic Taxonomy by utilizing morphologic and physiologic characters, and sequencing of ß-tubulin and calmodulin genes. The morphologic identification proved useful for some species, such as A. carbonarius and Aspergillus sp UFLA DCA 01, despite not having been totally effective in elucidating species related to A. niger. The isolation of the species of the Nigri Section on Creatine Sucrose Agar (CREA) enabled to distinguish the Aspergillus sp species, which was characterized by the lack of sporulation and by the production of sclerotia. Scanning Electron microscopy (SEM) allowed distinguishing the species into two distinct groups. The production of Ochratoxin A (OTA) was only found in the A. carbonarius and A. niger species. The sequencing of ?-tubulin gene was efficient in differing most of the Aspergillus species from the Nigri Section with the exception of Aspergillus UFLA DCA 01, which could not be distinguished from A. costaricaensis. This species is morphologically similar to A. costaricaencis for its low sporulation capacity and high sclerotia production, but it differs morphologically from A. costaricaensis for its conidial ornamentation and size of vesicles. Equally, based on partial calmodulin gene sequence data Aspergillus UFLA DCA 01 differs from A. costaricaensis. PMID:24031691

Silva, Daiani M.; Batista, Luís R.; Rezende, Elisângela F.; Fungaro, Maria Helena P.; Sartori, Daniele; Alves, Eduardo

2011-01-01

381

Occurrence of toxigenic Aspergillus spp. and aflatoxins in selected food commodities of Asian origin sourced in the West of Scotland.  

PubMed

The occurrence of Aspergillus moulds and aflatoxins in 12 commercially-available dried foods of Asian origin were examined. All food samples, except green beans and three types of dried fruit, contained multiple genera of moulds of which Aspergillus (55%) was the most frequently detected. Penicillium (15%), Rhizopus (11%), Mucor (3%), Monascus (1%), Eurotium (1%) and unidentified (14%) were also observed. The occurrence of aflatoxigenic moulds, however, did not correspond with the occurrence of aflatoxins in foods. Aflatoxigenic Aspergillus spp. (39 isolates) were recovered from long grain rice, fragrant rice, peanuts, black beans and black pepper. The predominant Aspergillus species was A. parasiticus (61%) while Aspergillus oryzae (3%), Aspergillus utus (5%), Aspergillus niger (5%), Aspergillus ochraceus (3%) and unidentified (23%) were also observed. Long grain rice, fragrant rice, peanuts, black beans and black pepper were positive for Aspergillus but contained undetectable aflatoxins. In contrast, Jasmine brown rice and crushed chilli contained 14.7 and 11.4?g/kg of total aflatoxins, respectively, in the absence of Aspergillus so aflatoxigenic Aspergillus was present at some stage of food production. The results from this study emphasise the need for stricter control measures in reducing occurrence of aflatoxins in foods for export and domestic use. PMID:23416649

Ruadrew, Sayan; Craft, John; Aidoo, Kofi

2013-05-01

382

Multilocus variable-number tandem-repeat analysis of clinical isolates of Aspergillus flavus from Iran reveals the first cases of Aspergillus minisclerotigenes associated with human infection  

PubMed Central

Background Aspergillus flavus is intensively studied for its role in infecting crop plants and contaminating produce with aflatoxin, but its role as a human pathogen is less well understood. In parts of the Middle East and India, A. flavus surpasses A. fumigatus as a cause of invasive aspergillosis and is a significant cause of cutaneous, sinus, nasal and nail infections. Methods A collection of 45 clinical and 10 environmental A. flavus isolates from Iran were analysed using Variable-Number Tandem-Repeat (VNTR) markers with MICROSAT and goeBURST to determine their genetic diversity and their relatedness to clinical and environmental A. flavus isolates from Australia. Phylogeny was assessed using partial ?-tubulin and calmodulin gene sequencing, and mating type was determined by PCR. Antifungal susceptibility testing was performed on selected isolates using a reference microbroth dilution method. Results There was considerable diversity in the A. flavus collection, with no segregation on goeBURST networks according to source or geographic location. Three Iranian isolates, two from sinus infections and one from a paranasal infection grouped with Aspergillus minisclerotigenes, and all produced B and G aflatoxin. Phylogenic analysis using partial ?-tubulin and calmodulin sequencing confirmed two of these as A. minisclerotigenes, while the third could not be differentiated from A. flavus and related species within Aspergillus section flavi. Based on epidemiological cut-off values, the A. minisclerotigens and A. flavus isolates tested were susceptible to commonly used antifungal drugs. Conclusions This is the first report of human infection due to A. minisclerotigenes, and it raises the possiblity that other species within Aspergillus section flavi may also cause clinical disease. Clinical isolates of A. flavus from Iran are not distinct from Australian isolates, indicating local environmental, climatic or host features, rather than fungal features, govern the high incidence of A. flavus infection in this region. The results of this study have important implications for biological control strategies that aim to reduce aflatoxin by the introduction of non-toxigenic strains, as potentially any strain of A. flavus, and closely related species like A. minisclerotigenes, might be capable of human infection. PMID:24986045

2014-01-01

383

Genetic Variability of Aspergillus flavus Isolates from a Mississippi Corn Field  

PubMed Central

A nontoxigenic Aspergillus flavus strain, K49, is currently being tested as a biological control agent in corn fields in the Mississippi Delta. However, little is known about the overall genetic diversity of A. flavus from year to year in corn fields and specifically in Mississippi. Our objective was to assess the genetic variability of A. flavus isolates from different seasons, inoculum sources, and years, from a no-till corn field. Of the 175 A. flavus isolates examined, 74 and 97 had the typical norB-cypA type I (1.5?kb) and type II (1.0?kb) deletion patterns, respectively. Variability in the sequence of the omtA gene of the majority of the field isolates (n = 118) was compared to strain K49. High levels of haplotypic diversity (24 omtA haplotypes; Hd = 0.61 ± 0.04) were found. Among the 24 haplotypes, two were predominant, H1 (n = 71), which consists of mostly toxigenic isolates, and H49 (n = 18), which consists of mostly atoxigenic isolates including K49. Toxigenic isolates were prevalent (60%) in this natural population. Nonetheless, about 15% of the population likely shared the same ancestral origin with K49. This study provides valuable information on the diversity of A. flavus. This knowledge can be further used to develop additional biological control strains. PMID:25478591

Solorzano, Cesar D.; Abbas, Hamed K.; Zablotowicz, Robert M.; Chang, Perng-Kuang; Jones, Walker A.

2014-01-01

384

Detection and discrimination of four Aspergillus section Nigri species by PCR.  

PubMed

Species of Aspergillus section Nigri are not easily distinguished by traditional morphological techniques, and typically are identified by DNA sequencing methods. We developed four PCR primers to distinguish between Aspergillus niger, Aspergillus welwitschiae, Aspergillus carbonarius and Aspergillus tubingensis, based on species-conserved differences in the calmodulin gene sequence. PCR amplification from total DNA using these primers was species specific; no amplification occurred from nontarget species DNA for each primer pair. Species-specific PCR could distinguish between species in mixed DNA templates, indicating a utility in determining culture uniformity of isolated Aspergillus strains. In addition, with these primer sets, each species could be detected in soil following mixed-species inoculation with Aspergillus spores. This indicates that PCR with these species-specific primers may be useful in determining the distribution of Aspergillus species in environmental samples without the need for species identification from isolated strains, as well as detecting species that may be infrequently isolated by culture-based methods. PMID:25384730

Palumbo, J D; O'Keeffe, T L

2015-02-01

385

Aspergillus citrinoterreus, a new species of section Terrei isolated from samples of patients with nonhematological predisposing conditions.  

PubMed

The use of molecular identification techniques has revealed an increasing number of new species within Aspergillus section Terrei. We phenotyped a set of 26 clinical isolates that showed genetic differences from Aspergillus terreus sensu stricto by analyzing sequences from PCR-amplified ?-tubulin and calmodulin genes and the internal transcribed spacer region. Since the isolates were phylogenetically and morphologically different from all of the members of Aspergillus section Terrei, they are described here as a new species, Aspergillus citrinoterreus, so named because it produces a diffusible yellowish pigment in agar. A. citrinoterreus isolates were significantly more susceptible to itraconazole, voriconazole, and posaconazole than A. terreus sensu stricto isolates were; in contrast, the amphotericin B MICs for both species were high. A. citrinoterreus was found in clinical samples from patients with proven or probable invasive aspergillosis and colonized patients, none of whom had hematological malignancies as predisposing conditions. However, they did have other underlying conditions such as chronic obstructive pulmonary disease, cirrhosis, and cancer or had received a solid organ transplants and presented not only with invasive pulmonary aspergillosis but also with mediastinitis. A. citrinoterreus isolates were detected for the first time in 2002. In all cases of invasive aspergillosis, A. citrinoterreus was found to be a copathogen, mostly with A. fumigatus. PMID:25502530

Guinea, Jesús; Sandoval-Denis, Marcelo; Escribano, Pilar; Peláez, Teresa; Guarro, Josep; Bouza, Emilio

2015-02-01

386

Penicillium parvulum and Penicillium georgiense, sp. nov., isolated from the conidial heads of Aspergillus species.  

PubMed

Two new Penicillium species were isolated from peanut-field soils in Georgia. The species were noted particularly because they sporulated on the conidial heads of Aspergillus species. Phenotypic descriptions were prepared with standard media. LSU-rDNA sequences were determined for the new species and compared to existing homologous sequences from Penicillium species with parsimony analysis. The monoverticillate species, P. parvulum, was related most closely to E. cinnamopurpureum, while the furcate species, P. georgiense, appeared in the tree near P. thiersii. Because P. parvulum was closely related to E. cinnamopurpureum additional loci were sequenced (beta-tubulin and calmodulin) for these and some other closely related species to establish the status of the species through genealogical concordance. Some proposed synonymies from prior studies were examined and resolved. PMID:19274850

Peterson, Stephen W; Horn, Bruce W

2009-01-01

387

Interpretation of mtDNA RFLP variability among Aspergillus tubingensis isolates.  

PubMed

Aspergillus tubingensis isolates collected from distant geographic areas were earlier classified into six groups on the basis of the mtDNA RFLP variability they exhibited (mtDNA types 2a-2f). In the present work, we investigated the reason for the intraspecific mtDNA variability and we describe here how this fungus, with a relatively small mitochondrial genome, can display intraspecific polymorphism due to intron acquisition and also sporadic point mutations affecting the recognition motifs of the restriction enzymes employed in the RFLP analysis. Three different LAGLI-DADG type group I introns were identified in the cox1 gene amongst the six mtDNA RFLP types. MtDNAs of types 2b and 2d contain all of the three introns, mtDNA of type 2f carries only one, and the other mtDNA types contain two introns each. Comparative analysis showed that the first and second introns of mtDNAs of types 2b and 2d are well distributed among fungi, indicating their active horizontal transfer capacity. The third intron occurs rarely among fungi and is restricted to a limited number of fungal species, namely to A. tubingensis and the yeast Candida stellata. It is interesting that this intron is present in a small mitochondrial genome such as that of A. tubingensis and, considering its rarity, its presence amongst black Aspergillus isolates is recommended to be considered as a tool to establish taxonomical unit(s) or to track down evolutionary divergence of closely related taxonomical units. PMID:17043909

Juhász, Akos; Engi, Helga; Pfeiffer, Ilona; Kucsera, Judit; Vágvölgyi, Csaba; Hamari, Zsuzsanna

2007-04-01

388

Characterization of Expressed Sequence Tag-Derived Simple Sequence Repeat Markers for Aspergillus flavus: Emphasis on Variability of Isolates from the Southern United States  

Technology Transfer Automated Retrieval System (TEKTRAN)

Simple Sequence Repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers wer...

389

Isolation, structure elucidation, and biomimetic total synthesis of versicolamide B and the isolation of antipodal (-)-stephacidin A and (+)-notoamide B from Aspergillus versicolor  

Technology Transfer Automated Retrieval System (TEKTRAN)

A new prenylated indole alkaloid, versicolamide B, was isolated from cultures of Aspergillus versicolor NRRL 35600. The structure was assigned by 2D NMR data, and confirmed by a biomimetic total synthesis. Versicolamide B is the first member of the paraherquamide-stephacidin family of alkaloids fo...

390

Occurrence of Toxigenic Aspergillus versicolor Isolates and Sterigmatocystin in Carpet Dust from Damp Indoor Environments  

PubMed Central

Over the past decade, there has been growing concern regarding the role of toxigenic fungi in damp indoor environments; however, there is still a lack of field investigations on exposure to mycotoxins. The goal of our pilot study was to quantify the proportion of toxigenic Aspergillus versicolor isolates in native carpet dust from damp dwellings with mold problems and to determine whether sterigmatocystin can be detected in this matrix. Carpet dust samples (n = 11) contained from <2.5 × 101 to 3.6 × 105 (median, 3.1 × 104) A. versicolor CFU/g of dust, and the median proportion of A. versicolor from total culturable fungi was 18%. Based on thin-layer chromatography detection of sterigmatocystin, 49 of 50 A. versicolor isolates (98%) were found to be toxigenic in vitro. By using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry, sterigmatocystin could be detected in low concentrations (2 to 4 ng/g of dust) in 2 of 11 native carpet dust samples. From this preliminary study, we conclude that most strains of A. versicolor isolated from carpet dust are able to produce sterigmatocystin in vitro and that sterigmatocystin may occasionally occur in carpet dust from damp indoor environments. Further research and systematic field investigation are needed to confirm our results and to provide an understanding of the health implications of mycotoxins in indoor environments. PMID:12147486

Engelhart, Steffen; Loock, Annette; Skutlarek, Dirk; Sagunski, Helmut; Lommel, Annette; Färber, Harald; Exner, Martin

2002-01-01

391

A single mutation in the hepta-peptide active site of Aspergillus niger PhyA phytase leads to myriad of biochemical changes  

Technology Transfer Automated Retrieval System (TEKTRAN)

The active site motif of proteins belonging to ‘Histidine Acid Phosphatase’ (HAP) contains a hepta-peptide region, RHGXRXP. A close comparison among fungal and yeast HAPs has revealed the fourth residue of the hepta-peptide to be E instead of A, which is the case with A. niger phyA phytase. However,...

392

Aspergillus fumigatus and related species.  

PubMed

The genus Aspergillus contains etiologic agents of aspergillosis. The clinical manifestations of the disease range from allergic reaction to invasive pulmonary infection. Among the pathogenic aspergilli, Aspergillus fumigatus is most ubiquitous in the environment and is the major cause of the disease, followed by Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus nidulans, and several species in the section Fumigati that morphologically resemble A. fumigatus. Patients that are at risk for acquiring aspergillosis are those with an altered immune system. Early diagnosis, species identification, and adequate antifungal therapy are key elements for treatment of the disease, especially in cases of pulmonary invasive aspergillosis that often advance very rapidly. Incorporating knowledge of the basic biology of Aspergillus species to that of the diseases that they cause is fundamental for further progress in the field. PMID:25377144

Sugui, Janyce A; Kwon-Chung, Kyung J; Juvvadi, Praveen R; Latgé, Jean-Paul; Steinbach, William J

2015-02-01

393

Multilocus sequence analysis of Aspergillus Sect. Nigri in dried vine fruits of worldwide origin.  

PubMed

Dried vine fruits may be heavily colonized by Aspergillus species. The molecular biodiversity of an Aspergillus population (234 strains) isolated from dried vine fruit samples of worldwide origin were analyzed by investigating four housekeeping gene loci (calmodulin, ?-tubulin, elongation factor 1-?, RPB2). Aspergillus Sect. Nigri was dominant and the strains were identified as A. tubingensis (138), A. awamori (38), A. carbonarius (27), A. uvarum (16) and A. niger (11). Four Aspergillus flavus strains were also identified from Chilean raisins. Two clusters closely related to the A. tubingensis species with a significant bootstrap (60% and 99%) were identified as distinct populations. Among the four loci, RPB2 showed the highest genetic variability. This is the first complete study on the worldwide distribution of black Aspergilli occurring on dried vine fruits identified by a molecular approach. PMID:23732831

Susca, Antonia; Perrone, Giancarlo; Cozzi, Giuseppe; Stea, Gaetano; Logrieco, Antonio F; Mulè, Giuseppina

2013-07-15

394

Drug resistance of Aspergillus fumigatus strains isolated from flocks of domestic geese in Poland.  

PubMed

The aim of the study was to determine the in vitro susceptibility of 85 Aspergillus fumigatus strains isolated from domestic geese and from their environment to amphotericin B, clotrimazole, voriconazole, itraconazole, enilconazole, miconazole, ketoconazole, and tioconazole. Samples were collected from clinically healthy birds (oral cavity) and from birds with aspergillosis (lungs and air sacs). The study was carried out using the disk diffusion method according to the Clinical Laboratory and Standards Institute (CLSI) M44-A2 procedure in parallel with the microdilution broth method according to CLSI M38-A2. The disk diffusion method showed that the all of the strains, irrespective of source, were resistant to miconazole. Resistance to the remaining azoles and amphotericin B ranged from 90.6 to 70.6%. Complete susceptibility was noted for voriconazole and enilconazole. Determination of the minimum inhibitory concentration (MIC) confirmed the high resistance of the strains tested to clotrimazole (MIC90 = 16 µg•mL(-1)), amphotericin B (MIC90 = 16 µg•mL(-1)), varied susceptibility to itraconazole (MIC 0.5-8 µg•mL(-1)), and 100% susceptibility to enilconazole and voriconazole. A correlation was noted between the susceptibility of the strains and their source. The highest percentage of resistant strains was noted in isolates from the lungs (100% for amphotericin B and clotrimazole and 35.7% for itraconazole). To the best of our knowledge, this is the first monitoring conducted in Poland in this area of research. PMID:24795302

Zio?kowska, Grazyna; Tokarzewski, Stanis?aw; Nowakiewicz, Aneta

2014-05-01

395

Physiological behaviour of gliotoxigenic Aspergillus fumigatus sensu stricto isolated from maize silage under simulated environmental conditions.  

PubMed

Environmental conditions play a key role in fungal development. During the silage production process, humidity, oxygen availability and pH vary among lactic-fermentation phases and among different silage sections. The aim of this work was to study the physiological behaviour of gliotoxicogenic Aspergillus fumigatus strains isolated from maize silage under simulated natural physicochemical conditions - different water activities (a(W)), temperatures (Tº), pH and oxygen pressure - on the growth parameters (growth rate and lag phase) and gliotoxin production. The silage was made with the harvested whole maize plant that was chopped and used for trench-type silo fabrication. Water activity and pH of the silage samples were determined. Total fungal counts were performed on Dichloran Rose Bengal Chloramphenicol agar and Dichloran 18% Glycerol agar. The morphological identification of A. fumigatus was performed with different culture media and at different growth temperature to observe microscopic and macroscopic characteristics. Gliotoxin production by A. fumigatus was determined by HPLC. All strains isolated were morphologically identified as A. fumigatus. Two A. fumigatus strains isolated from the silage samples were selected for the ecophysiological study (A. fumigatus sensu stricto RC031 and RC032). The results of this investigation showed that the fungus grows in the simulated natural physicochemical conditions of corn silage and produces gliotoxin. The study of the physiological behaviour of gliotoxigenic A. fumigatus under simulated environmental conditions allowed its behaviour to be predicted in silage and this will in future enable appropriate control strategies to be developed to prevent the spread of this fungus and toxin production that leads to impairment and reduced quality of silage. PMID:25599419

Alonso, V; Vergara, L Díaz; Aminahuel, C; Pereyra, C; Pena, G; Torres, A; Dalcero, A; Cavaglieri, L

2015-01-01

396

Indole Diterpenoids and Isocoumarin from the Fungus, Aspergillus flavus, Isolated from the Prawn, Penaeus vannamei  

PubMed Central

Two new indole-diterpenoids (1 and 2) and a new isocoumarin (3), along with the known ?-aflatrem (4), paspalinine (5), leporin B (6), ?-cyclopiazonic acid (7), iso-?-cyclopiazonic acid (8), ditryptophenaline (9), aflatoxin B1 (10), 7-O-acetylkojic acid (11) and kojic acid (12), were isolated from the fermentation broth of the marine-derived fungus, Aspergillus flavus OUCMDZ-2205. The structures of Compounds 1–12 were elucidated by spectroscopic analyses, quantum ECD calculations and the chemical method. New Compound 1 exhibited antibacterial activity against Staphylococcus aureus with a MIC value of 20.5 ?M. Both new Compounds 1 and 2 could arrest the A549 cell cycle in the S phase at a concentration of 10 ?M. Compound 1 showed PKC-beta inhibition with an IC50 value of 15.6 ?M. In addition, the absolute configurations of the known compounds, 4–6 and leporin A (6a), were also determined for the first time. PMID:24983640

Sun, Kunlai; Li, Ye; Guo, Lei; Wang, Yi; Liu, Peipei; Zhu, Weiming

2014-01-01

397

Isolation and characterisation of the acetyl-CoA carboxylase gene from Aspergillus nidulans.  

PubMed

The putative gene encoding acetyl-CoA carboxylase, accA, has been isolated from Aspergillus nidulans. This single-copy gene has an open reading frame (ORF) of 6864 bp and contains two small introns near the 5'-end. A short ORF upstream of the ATG start codon has been identified in this gene by RT-PCR. Based on sequence homology to acetyl-CoA carboxylases from other organisms, putative biotin-, ATP-, HCO3-- and acetyl-CoA- binding sites have been assigned. Northern data and ACC enzyme-activity measurements from A. nidulans suggested that expression of accA was higher in media containing nitrate than ammonia as a sole nitrogen source. Deletion of accA in A. nidulans was unsuccessful. The failure of A. nidulans to grow in the presence of the ACC-specific inhibitor, soraphen A, supplemented with C16-18 fatty acids suggested that ACC is an essential enzyme. PMID:9871120

Morrice, J; MacKenzie, D A; Parr, A J; Archer, D B

1998-12-01

398

Enfumafungin derivative MK-3118 shows increased in vitro potency against clinical echinocandin-resistant Candida Species and Aspergillus species isolates.  

PubMed

MK-3118 is as an orally active new antifungal in the early stage of clinical development that inhibits the biosynthesis of ?-(1,3)-glucan. We evaluated the in vitro activity of this compound against wild-type and echinocandin-resistant (ER) isolates containing mutations in the FKS gene(s) of Candida spp. and Aspergillus spp. MK-3118 demonstrated enhanced efficacy for most C. albicans and C. glabrata ER isolates relative to caspofungin, with decreased MICs and half-maximal inhibitory concentrations (IC50s). PMID:24323472

Jiménez-Ortigosa, Cristina; Paderu, Padmaja; Motyl, Mary R; Perlin, David S

2014-01-01

399

Determination of isavuconazole susceptibility of Aspergillus and Candida species by the EUCAST method.  

PubMed

Isavuconazole is a novel expanded-spectrum triazole, which has recently been approved by the FDA as an orphan drug to treat invasive aspergillosis and is currently being studied in phase III clinical trials for invasive candidiasis. The susceptibility of relatively few clinical isolates has been reported. In this study, the isavuconazole susceptibilities of 1,237 Aspergillus and 2,010 Candida geographically diverse clinical isolates were determined by EUCAST methodology at four European mycology laboratories, producing the largest multicenter data set thus far for this compound. In addition, a blinded collection of 30 cyp51A mutant Aspergillus fumigatus clinical isolates and 10 wild-type isolates was tested. From these two data sets, the following preliminary epidemiological cutoff (ECOFF) values were suggested: 2 mg/liter for Aspergillus fumigatus, Aspergillus terreus, and Aspergillus flavus; 4 mg/liter for Aspergillus niger; 0.25 mg/liter for Aspergillus nidulans; and 0.03 mg/liter for Candida albicans, Candida parapsilosis, and Candida tropicalis. Unfortunately, ECOFFs could not be determined for Candida glabrata or Candida krusei due to an unexplained interlaboratory MIC variation. For the blinded collection of A. fumigatus isolates, all MICs were ?2 mg/liter for wild-type isolates. Differential isavuconazole MICs were observed for triazole-resistant A. fumigatus isolates with different cyp51A alterations: TR34/L98H mutants had elevated isavuconazole MICs, whereas isolates with G54 and M220 alterations had MICs in the wild-type range, suggesting that the efficacy of isavuconazole may not be affected by these alterations. This study will be an aid in interpreting isavuconazole MICs for clinical care and an important step in the future process of setting official clinical breakpoints. PMID:23959309

Howard, Susan J; Lass-Flörl, Cornelia; Cuenca-Estrella, Manuel; Gomez-Lopez, Alicia; Arendrup, Maiken C

2013-11-01

400

Determination of Isavuconazole Susceptibility of Aspergillus and Candida Species by the EUCAST Method  

PubMed Central

Isavuconazole is a novel expanded-spectrum triazole, which has recently been approved by the FDA as an orphan drug to treat invasive aspergillosis and is currently being studied in phase III clinical trials for invasive candidiasis. The susceptibility of relatively few clinical isolates has been reported. In this study, the isavuconazole susceptibilities of 1,237 Aspergillus and 2,010 Candida geographically diverse clinical isolates were determined by EUCAST methodology at four European mycology laboratories, producing the largest multicenter data set thus far for this compound. In addition, a blinded collection of 30 cyp51A mutant Aspergillus fumigatus clinical isolates and 10 wild-type isolates was tested. From these two data sets, the following preliminary epidemiological cutoff (ECOFF) values were suggested: 2 mg/liter for Aspergillus fumigatus, Aspergillus terreus, and Aspergillus flavus; 4 mg/liter for Aspergillus niger; 0.25 mg/liter for Aspergillus nidulans; and 0.03 mg/liter for Candida albicans, Candida parapsilosis, and Candida tropicalis. Unfortunately, ECOFFs could not be determined for Candida glabrata or Candida krusei due to an unexplained interlaboratory MIC variation. For the blinded collection of A. fumigatus isolates, all MICs were ?2 mg/liter for wild-type isolates. Differential isavuconazole MICs were observed for triazole-resistant A. fumigatus isolates with different cyp51A alterations: TR34/L98H mutants had elevated isavuconazole MICs, whereas isolates with G54 and M220 alterations had MICs in the wild-type range, suggesting that the efficacy of isavuconazole may not be affected by these alterations. This study will be an aid in interpreting isavuconazole MICs for clinical care and an important step in the future process of setting official clinical breakpoints. PMID:23959309

Howard, Susan J.; Lass-Flörl, Cornelia; Cuenca-Estrella, Manuel; Gomez-Lopez, Alicia

2013-01-01

401

Aspergillus luchuensis, an industrially important black Aspergillus in East Asia.  

PubMed

Aspergilli known as black- and white-koji molds which are used for awamori, shochu, makgeolli and other food and beverage fermentations, are reported in the literature as A. luchuensis, A. awamori, A. kawachii, or A. acidus. In order to elucidate the taxonomic position of these species, available ex-type cultures were compared based on morphology and molecular characters. A. luchuensis, A. kawachii and A. acidus showed the same banding patterns in RAPD, and the three species had the same rDNA-ITS, ?-tubulin and calmodulin sequences and these differed from those of the closely related A. niger and A. tubingensis. Morphologically, the three species are not significantly different from each other or from A. niger and A. tubingensis. It is concluded that A. luchuensis, A. kawachii and A. acidus are the same species, and A. luchuensis is selected as the correct name based on priority. Strains of A. awamori which are stored in National Research Institute of Brewing in Japan, represent A. niger (n?=?14) and A. luchuensis (n?=?6). The neotype of A. awamori (CBS 557.65?=? NRRL 4948) does not originate from awamori fermentation and it is shown to be identical with the unknown taxon Aspergillus welwitschiae. Extrolite analysis of strains of A. luchuensis showed that they do not produce mycotoxins and therefore can be considered safe for food and beverage fermentations. A. luchuensis is also frequently isolated from meju and nuruk in Korea and Puerh tea in China and the species is probably common in the fermentation environment of East Asia. A re-description of A. luchuensis is provided because the incomplete data in the original literature. PMID:23723998

Hong, Seung-Beom; Lee, Mina; Kim, Dae-Ho; Varga, Janos; Frisvad, Jens C; Perrone, Giancarlo; Gomi, Katsuya; Yamada, Osamu; Machida, Masayuki; Houbraken, Jos; Samson, Robert A

2013-01-01

402

Isolation and characterization of the Aspergillus parasiticus pacC gene  

E-print Network

and carcinogenic secondary metabolites produced by several Aspergi llus species. Ambient pH has been determined to affect mycotoxin (i. e. aflatoxin and sterigmatocystin) biosynthesis in Aspergillus parasiticus and Aspergillus nidulans, respectively.... The construction of constitutive mutations of pacC is achieved by engineering a truncated protein, which acts as the activated form. This permits an additional level of investigation into the effects of PacC and pH regulation besides that of a null mutation...

Pinero, David

1999-01-01

403

Four butyrolactones and diverse bioactive secondary metabolites from terrestrial Aspergillus flavipes MM2: isolation and structure determination  

PubMed Central

The chemical constituents and biological activities of the terrestrial Aspergillus flavipes MM2 isolated from Egyptian rice hulls are reported. Seven bioactive compounds were obtained, of which one sterol: ergosterol (1), four butyrolactones: butyrolactone I (2), aspulvinone H (3), butyrolactone-V (6) and 4,4'-diydroxypulvinone (7), along with 6-methylsalicylic acid (4) and the cyclopentenone analogue; terrien (5). Structures of the isolated compounds were deduced by intensive studies of their 1D & 2D NMR, MS data and comparison with related structures. The strain extract and the isolated compounds (1-7) were biologically studied against number of microbial strains, and brine shrimp for cytotoxicity. In this article, the taxonomical characterization of A. flavipes MM2 along with its upscale fermentation, isolation and structural assignment of the obtained bioactive metabolites, and evaluate their antimicrobial and cytotoxic activities were described. PMID:22380482

2012-01-01

404

Three-dimensional Structure of Endo-1,4-?-xylanase I from Aspergillus niger : Molecular Basis for its Low pH Optimum  

Microsoft Academic Search

The crystal structure of endo-1,4-?-xylanase I fromAspergillus nigerhas been solved by molecular replacement and was refined to 2.4 Å resolution. The finalR-factor for all data from 6 to 2.4 Å is 17.9%. TheA. nigerxylanase has a characteristic fold which is unique for family G xylanases (root-mean-square deviation=1.1 Å toTrichoderma reeseixylanase I, which has 53% sequence identity). It consists of a

Ute Krengel; Bauke W. Dijkstra

1996-01-01

405

Study of anti-inflammatory, analgesic and antipyretic activities of seeds of Hyoscyamus niger and isolation of a new coumarinolignan  

Microsoft Academic Search

A chemical and biological validation of the traditional use of Hyoscyamus niger seeds as anti-inflammatory drug has been established. The methanolic extract of seeds of H. niger (MHN) was evaluated for its analgesic, anti-inflammatory and antipyretic activities in experimental animal models at different doses. MHN produced significant increase in hot plate reaction time, while decreasing writhing response in a dose-dependent

Sajeli Begum; Bhagawati Saxena; Madhur Goyal; Rakesh Ranjan; Vijaya B. Joshi; Ch V. Rao; Sairam Krishnamurthy; Mahendra Sahai

2010-01-01

406

Metabolism of citric acid production by Aspergillus niger: model definition, steady-state analysis and constrained optimization of citric acid production rate.  

PubMed

In an attempt to provide a rational basis for the optimization of citric acid production by A. niger, we developed a mathematical model of the metabolism of this filamentous fungus when in conditions of citric acid accumulation. The present model is based in a previous one, but extended with the inclusion of new metabolic processes and updated with currently available kinetic data. Among the different alternatives to represent the system behavior we have chosen the S-system representation within power-law formalism. This type of representation allows us to verify not only the ability of the model to exhibit a stable steady state of the integrated system but also the robustness and quality of the representation. The model analysis is shown to be self-consistent, with a stable steady state, and in good agreement with experimental evidence. Moreover, the model representation is sufficiently robust, as indicated by sensitivity and steady-state and dynamic analyses. From the steady-state results we concluded that the range of accuracy of the S-system representation is wide enough to model realistic deviations from the nominal steady state. The dynamic analysis indicated a reasonable response time, which provided further indication that the model is adequate. The extensive assessment of the reliability and quality of the model put us in a position to address questions of optimization of the system with respect to increased citrate production. We carried out the constrained optimization of A. niger metabolism with the goal of predicting an enzyme activity profile yielding the maximum rate of citrate production, while, at the same time, keeping all enzyme activities within predetermined, physiologically acceptable ranges. The optimization is based on a method described and tested elsewhere that utilizes the fact that the S-system representation of a metabolic system becomes linear at steady state, which allows application of linear programming techniques. Our results show that: (i) while the present profile of enzyme activities in A. niger at idiophase steady state yields high rates of citric acid production, it still leaves room for changes and suggests possible optimization of the activity profile to over five times the basal rate synthesis; (ii) when the total enzyme concentration is allowed to double its basal value, the citric acid production rate can be increased by more than 12-fold, and even larger values can be attained if the total enzyme concentration is allowed to increase even more (up to 50-fold when the total enzyme concentration may rise up to 10-fold the basal value); and (iii) the systematic search of the best combination of subsets of enzymes shows that, under all conditions assayed, a minimum of 13 enzymes need be modified if significant increases in citric acid are to be obtained. This implies that improvements by single enzyme modulation are unlikely, which is in agreement with the findings of some investigators in this and other fields. PMID:10940866

Alvarez-Vasquez, F; González-Alcón, C; Torres, N V

2000-10-01

407

Utilization of b-glucosidase from aspergillus species in the hydrolysis of cellulose  

SciTech Connect

The batch hydrolysis of cellulose by Trichoderma reesei cellulase was considerably enhanced by the addition of very small amounts of B-glucosidase derived from Aspergillus niger. Addition of larger amounts had no further effect. In simultaneous cellulose hydrolysis and alcohol fermentation experiments the addition of B-glucosidase from Aspergillus niger had no significant effect on alcohol production by the fermenting yeast.

Nybergh, P.M.A.; Bailey, M.J.

1980-01-01

408

Genetic variability of Aspergillus flavus isolates from a Mississippi corn field  

Technology Transfer Automated Retrieval System (TEKTRAN)

The fungus Aspergillus flavus represents a major threat to food safety and food security on a worldwide scale. Corn, peanuts, cotton, rice and edible nuts, can be colonized by A. flavus strains that produce carcinogenic aflatoxins. A biological strategy for control of toxigenic A. flavus starins inv...

409

Identification of atoxigenic Aspergillus flavus isolates to reduce aflatoxin contamination of maize in Kenya  

Technology Transfer Automated Retrieval System (TEKTRAN)

Acute aflatoxin poisonings (aflatoxicosis) in Kenya have led to the deaths of several hundred people between 2004 and 2006. Etiology of contamination in the outbreak districts (Eastern Province) identified an unusual fungal community structure dominated by the highly toxigenic Aspergillus flavus S s...

410

Genetic Isolation among Sympatric Vegetative Compatibility Groups of the aflatoxin-producing fungus Aspergillus flavus  

Technology Transfer Automated Retrieval System (TEKTRAN)

Aspergillus flavus, fungal pathogen of animals and both wild and economically important plants, is most recognized for producing aflatoxin, a cancer-causing secondary metabolite, that contaminates food and animal feed globally. A. flavus is asexual and has a vegetative incompatibility system that li...

411

Isolation of Aspergillus fumigatus from sputum is associated with elevated airborne levels in homes of patients with asthma.  

PubMed

Indoor bioaerosols, such as mold spores, have been associated with respiratory symptoms in patients with asthma; however, dose-response relationships and guidelines on acceptable levels are lacking. Furthermore, a causal link between mold exposure and respiratory infections or asthma remains to be established. The aim of this study was to determine indoor concentrations of Aspergillus fumigatus and a subset of clinically relevant fungi in homes of people with asthma, in relation to markers of airways colonization and sensitization. Air and dust samples were collected from the living room of 58 properties. Fungal concentrations were quantified using mold-specific quantitative PCR and compared with traditional microscopic analysis of air samples. Isolation of A. fumigatus from sputum was associated with higher airborne concentrations of the fungus in patient homes (P = 0.04), and a similar trend was shown with Aspergillus/Penicillium-type concentrations analyzed by microscopy (P = 0.058). No association was found between airborne levels of A. fumigatus and sensitization to this fungus, or dustborne levels of A. fumigatus and either isolation from sputum or sensitization. The results of this study suggest that the home environment should be considered as a potential source of fungal exposure, and elevated home levels may predispose people with asthma to airways colonization. PMID:23198683

Fairs, A; Agbetile, J; Bourne, M; Hargadon, B; Monteiro, W R; Morley, J P; Edwards, R E; Wardlaw, A J; Pashley, C H

2013-08-01

412

Biosynthesis and hyper production of pullulan by a newly isolated strain of Aspergillus japonicus-VIT-SB1.  

PubMed

The main focus of this study was to screen and characterize novel microbial strains isolated from culinary leaf samples, capable of producing high concentrations of pullulan. Hundred isolates were screened from the phylloplane of different plants. The results revealed that eight strains had the capability to produce exopolysaccharide (EPS) and only one potential strain (designated as VIT-SB1) could produce the significant amount of EPS (3.9 ± 0.02%) on the 6th day of the fermentation without optimisation. The EPS synthesized by VIT-SB1 strain was confirmed to be pullulan on the basis of the results of FT-IR, HPLC and the enzymatic (Pullulanase) analysis. More than 91% hydrolysis of pullulan by pullulanase enzyme also indicated the presence of ? (1 ? 6) glycosidic linkages of ? (1 ? 4) linked maltotriose units. This VIT-SB1 strain was identified as Aspergillus japonicus based on the nucleotide sequence of the D1/D2 domain of Large-Subunit rRNA gene. The sequence was submitted to the GenBank Nucleotide sequence database with Accession No: KC128815. This study has confirmed that pullulan production capacity of this novel strain and Aureobasidium pullulans are comparable. Hence Aspergillus japonicus-VIT-SB1 strain can be commercially exploited as a potential pullulan producing strain. PMID:24609496

Mishra, Bishwambhar; Suneetha, V

2014-07-01

413

Genome sequence comparison of Aspergillus fumigatus strains isolated from patients with pulmonary aspergilloma and chronic necrotizing pulmonary aspergillosis.  

PubMed

Aspergillus fumigatus is the Aspergillus species most commonly associated with aspergillosis. Of the various presentations of aspergillosis, one of the most frequently observed in cases involving A. fumigatus pulmonary infections is aspergilloma (PA). In such infections one finds a fungus ball composed of fungal hyphae, inflammatory cells, fibrin, mucus, and tissue debris. Chronic necrotizing pulmonary aspergillosis (CNPA), also known as semi-invasive or invasive aspergillosis, is locally invasive and predominantly seen in patients with mild immunodeficiency or with a chronic lung disease. In the present study, with the aid of a next-generation sequencer, we conducted whole genome sequence (WGS) analyses of 17 strains isolated from patients in Japan with PA and CNPA. A total of 99,088 SNPs were identified by mapping the reads to A. fumigatus genome reference strain Af293, and according to genome-wide phylogenetic analysis, there were no correlations between the whole genome sequence typing results and pathologic conditions of patients. Here, we conducted the first multi-genome WGS study to focus on the A. fumigatus strains isolated from patients with PA and CNPA, and comprehensively characterized genetic variations of strains. WGS approach will help in better understanding of molecular mechanisms of aspergillosis cases caused by A. fumigatus. PMID:25851262

Takahashi-Nakaguchi, Azusa; Muraosa, Yasunori; Hagiwara, Daisuke; Sakai, Kanae; Toyotome, Takahito; Watanabe, Akira; Kawamoto, Susumu; Kamei, Katsuhiko; Gonoi, Tohru; Takahashi, Hiroki

2015-05-01

414

Rapid Differentiation of Aspergillus Species from Other Medically Important Opportunistic Molds and Yeasts by PCR-Enzyme Immunoassay  

Microsoft Academic Search

We developed a PCR-based assay to differentiate medically important species of Aspergillus from one another and from other opportunistic molds and yeasts by employing universal, fungus-specific primers and DNA probes in an enzyme immunoassay format (PCR-EIA). Oligonucleotide probes, directed to the internal transcribed spacer 2 region of ribosomal DNA from Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus,

Liliana de Aguirre; Steven F. Hurst; Jong Soo Choi; Jong Hee Shin; Hans Peter Hinrikson; Christine J. Morrison

2004-01-01

415

Detection of Aspergillus-specific antibodies by agar gel double immunodiffusion and IgG ELISA in feline upper respiratory tract aspergillosis.  

PubMed

Feline upper respiratory tract aspergillosis (URTA) is an emerging infectious disease. The aims of this study were: (1) to assess the diagnostic value of detection of Aspergillus-specific antibodies using an agar gel double immunodiffusion (AGID) assay and an indirect immunoglobulin G (IgG) ELISA; and (2) to determine if an aspergillin derived from mycelia of Aspergillus fumigatus, Aspergillus niger and Aspergillus flavus can be used to detect serum antibodies against cryptic Aspergillus spp. in Aspergillus section Fumigati. Sera from cats with URTA (group 1: n?=?21) and two control groups (group 2: cats with other upper respiratory tract diseases, n?=?25; group 3: healthy cats and cats with non-respiratory, non-fungal illness, n?=?84) were tested. Isolates from cats with URTA comprised A. fumigatus (n?=?5), A. flavus (n?=?1) and four cryptic species: Aspergillus felis (n?=?12), Aspergillus thermomutatus (Neosartorya pseudofischeri, n?=?1), Aspergillus lentulus (n?=?1) and Aspergillus udagawae (n?=?1). Brachycephalic purebred cats were significantly more likely to develop URTA than other breeds (P?=?0.013). The sensitivity (Se) of the AGID was 43% and the specificity (Sp) was 100%. At a cut-off value of 6 ELISA units/mL, the Se of the IgG ELISA was 95.2% and the Sp was 92% and 92.9% for groups 2 and 3 cats, respectively. Aspergillus-specific antibodies against all four cryptic species were detected in one or both assays. Assay Se was not associated with species identity. Detection of Aspergillus-specific antibodies by IgG ELISA has high Se and Sp for diagnosis of feline URTA. PMID:25634077

Barrs, V R; Ujvari, B; Dhand, N K; Peters, I R; Talbot, J; Johnson, L R; Billen, F; Martin, P; Beatty, J A; Belov, K

2015-03-01

416

Multicenter study of isavuconazole MIC distributions and epidemiological cutoff values for Aspergillus spp. for the CLSI M38-A2 broth microdilution method.  

PubMed

Epidemiological cutoff values (ECVs) were established for the new triazole isavuconazole and Aspergillus species wild-type (WT) MIC distributions (organisms in a species-drug combination with no detectable acquired resistance mechanisms) that were defined with 855 Aspergillus fumigatus, 444 A. flavus, 106 A. nidulans, 207 A. niger, 384 A. terreus, and 75 A. versicolor species complex isolates; 22 Aspergillus section Usti isolates were also included. CLSI broth microdilution MIC data gathered in Europe, India, Mexico, and the United States were aggregated to statistically define ECVs. ECVs were 1 ?g/ml for the A. fumigatus species complex, 1 ?g/ml for the A. flavus species complex, 0.25 ?g/ml for the A. nidulans species complex, 4 ?g/ml for the A. niger species complex, 1 ?g/ml for the A. terreus species complex, and 1 ?g/ml for the A. versicolor species complex; due to the small number of isolates, an ECV was not proposed for Aspergillus section Usti. These ECVs may aid in detecting non-WT isolates with reduced susceptibility to isavuconazole due to cyp51A (an A. fumigatus species complex resistance mechanism among the triazoles) or other mutations. PMID:23716059

Espinel-Ingroff, A; Chowdhary, A; Gonzalez, G M; Lass-Flörl, C; Martin-Mazuelos, E; Meis, J; Peláez, T; Pfaller, M A; Turnidge, J

2013-08-01

417

Multicenter Study of Isavuconazole MIC Distributions and Epidemiological Cutoff Values for Aspergillus spp. for the CLSI M38-A2 Broth Microdilution Method  

PubMed Central

Epidemiological cutoff values (ECVs) were established for the new triazole isavuconazole and Aspergillus species wild-type (WT) MIC distributions (organisms in a species-drug combination with no detectable acquired resistance mechanisms) that were defined with 855 Aspergillus fumigatus, 444 A. flavus, 106 A. nidulans, 207 A. niger, 384 A. terreus, and 75 A. versicolor species complex isolates; 22 Aspergillus section Usti isolates were also included. CLSI broth microdilution MIC data gathered in Europe, India, Mexico, and the United States were aggregated to statistically define ECVs. ECVs were 1 ?g/ml for the A. fumigatus species complex, 1 ?g/ml for the A. flavus species complex, 0.25 ?g/ml for the A. nidulans species complex, 4 ?g/ml for the A. niger species complex, 1 ?g/ml for the A. terreus species complex, and 1 ?g/ml for the A. versicolor species complex; due to the small number of isolates, an ECV was not proposed for Aspergillus section Usti. These ECVs may aid in detecting non-WT isolates with reduced susceptibility to isavuconazole due to cyp51A (an A. fumigatus species complex resistance mechanism among the triazoles) or other mutations. PMID:23716059

Chowdhary, A.; Gonzalez, G. M.; Lass-Flörl, C.; Martin-Mazuelos, E.; Meis, J.; Peláez, T.; Pfaller, M. A.; Turnidge, J.

2013-01-01

418

Volatile profiles and aflatoxin production by toxigenic and non-toxigenic isolates of Aspergillus flavus grown on sterile and non-sterile cracked corn  

Technology Transfer Automated Retrieval System (TEKTRAN)

Aspergillus flavus is a saprophytic fungus which can grow on corn and produce aflatoxins which render it unsafe for food and feed consumption. In this study, aflatoxin and non-aflatoxin producing isolates of A. flavus were grown separately on wet (20% water added), sterile or non-sterile cracked co...

419

Identification of Aspergillus species using internal transcribed spacer regions 1 and 2.  

PubMed

Aspergillus species are the most frequent cause of invasive mold infections in immunocompromised patients. Although over 180 species are found within the genus, 3 species, Aspergillus flavus, A. fumigatus, and A. terreus, account for most cases of invasive aspergillosis (IA), with A. nidulans, A. niger, and A. ustus being rare causes of IA. The ability to distinguish between the various clinically relevant Aspergillus species may have diagnostic value, as certain species are associated with higher mortality and increased virulence and vary in their resistance to antifungal therapy. A method to identify Aspergillus at the species level and differentiate it from other true pathogenic and opportunistic molds was developed using the 18S and 28S rRNA genes for primer binding sites. The contiguous internal transcribed spacer (ITS) region, ITS 1-5.8S-ITS 2, from referenced strains and clinical isolates of aspergilli and other fungi were amplified, sequenced, and compared with non-reference strain sequences in GenBank. ITS amplicons from Aspergillus species ranged in size from 565 to 613 bp. Comparison of reference strains and GenBank sequences demonstrated that both ITS 1 and ITS 2 regions were needed for accurate identification of Aspergillus at the species level. Intraspecies variation among clinical isolates and reference strains was minimal. Sixteen other pathogenic molds demonstrated less than 89% similarity with Aspergillus ITS 1 and 2 sequences. A blind study of 11 clinical isolates was performed, and each was correctly identified. Clinical application of this approach may allow for earlier diagnosis and selection of effective antifungal agents for patients with IA. PMID:10747135

Henry, T; Iwen, P C; Hinrichs, S H

2000-04-01

420

Characterization of expressed sequence tag-derived simple sequence repeat markers for Aspergillus flavus: emphasis on variability of isolates from the southern United States.  

PubMed

Simple sequence repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers were designed from 362 tri-nucleotide EST-SSR sequences. Eighteen polymorphic loci were used to genotype 96 Aspergillus species isolates. The number of alleles detected per locus ranged from 2 to 24 with a mean of 8.2 alleles. Haploid diversity ranged from 0.28 to 0.91. Genetic distance matrix was used to perform principal coordinates analysis (PCA) and to generate dendrograms using unweighted pair group method with arithmetic mean (UPGMA). Two principal coordinates explained more than 75 % of the total variation among the isolates. One clade was identified for A. flavus isolates (n = 87) with the other Aspergillus species (n = 7) using PCA, but five distinct clusters were present when the others taxa were excluded from the analysis. Six groups were noted when the EST-SSR data were compared using UPGMA. However, the latter PCA or UPGMA comparison resulted in no direct associations with host species, geographical region or aflatoxin production. Furthermore, there was no direct correlation to visible morphological features such as sclerotial types. The isolates from Mississippi Delta region, which contained the largest percentage of isolates, did not show any unusual clustering except for isolates K32, K55, and 199. Further studies of these three isolates are warranted to evaluate their pathogenicity, aflatoxin production potential, additional gene sequences (e.g., RPB2), and morphological comparisons. PMID:22911544

Wang, Xinwang; Wadl, Phillip A; Wood-Jones, Alicia; Windham, Gary; Trigiano, Robert N; Scruggs, Mary; Pilgrim, Candace; Baird, Richard

2012-12-01

421

Effect of acid lactic bacteria isolated from faeces of healthy dogs on growth parameters and aflatoxin B 1 production by Aspergillus species in vitro  

Microsoft Academic Search

The aim of the present study was to evaluate the inhibitory effect of Enterococcus faecium and Lactococcus lactis subsp. lactis isolated from faeces of healthy dogs on (i) lag phase, (ii) growth rate, and (iii) aflatoxin B1 production by Aspergillus section Flavi on in vitro assays. Thirteen lactic acid bacteria (LAB) isolates were used as antagonist microorganisms. Antagonistic activity\\u000a was

María Guillermina Fernández-Juri; Julián A. Muzzolón; Ana María Dalcero; Carina E. Magnoli

422

[Isolation and characterization of the ALP1 protease from Aspergillus fumigatus and its protein inhibitor from Physarium polycephalum].  

PubMed

It is known that Aspergillus fumigatus secretes a serine protease ALP1 of the subtilisin family in the presence of extracellular protein substrates. We found conditions of A. fumigatus culturing that provide a high ALP1 activity inside cells without induction by extracellular proteins. The identity of the properties of the secreted and intracellular enzymes was shown. A thermostable protein inhibitor of the ALP1 protease was isolated from the plasmodium of the myxomycete Physarum polycephalum. Its molecular mass is 32-33 kDa. The inhibitor inhibits the ALP1 protease activity with IC50 of 0.14 microM. This protein was also shown to be a less efficient inhibitor of the activity of HIV-1 protease (IC50 2.5 microM). The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru. PMID:16004384

Davies, D A; Kalinina, N A; Samokhvalova, L V; Malakhova, G V; Scott, G; Volynskaia, A M; Nesmeianov, V A

2005-01-01

423

Mycotoxicosis produced in swine by cultural products of an isolate of Aspergillus ochraceus. II. Clinicopathologic changes.  

PubMed

Rice culture of a toxigenic strain of Aspergillus ochraceus was fed at a concentration of 25% to weanling pigs for 10 days. The clinicopathological abnormalities reflected renal damage. Activities of lactic dehydrogenase, glutamic-oxaloacetic transaminase and isocitrate dehydrogenase were increased in the urine but not in the serum. Serum concentrations of urea nitrogen and creatinine were high. Cellular and granular casts, blood, protein, and glucose were in the urine of pigs fed toxic diet. Serum concentrations of K+, Na+ and Cl- were unchanged, but concentrations of these electrolytes were reduced in the urine. PMID:505895

Zimmermann, J L; Carlton, W W; Tuite, J

1979-11-01

424

Comparative Evaluation of Disc Diffusion and E-test with Broth Micro-dilution in Susceptibility testing of Amphotericin B, Voriconazole and Caspofungin against Clinical Aspergillus isolates  

PubMed Central

Background: Clinical importance of Aspergillus has increased over the past few decades because of rise in immunosuppressive drugs and immune-modulating diseases. Antifungal susceptibility of Aspergillus is rarely performed by clinical laboratories because of lack of easier method. This study has investigated and compared susceptibility pattern of Aspergillus isolates by disc diffusion, E-test and broth micro-dilution for amphotericin B, voriconazole and caspofungin. Materials and Methods: Disk diffusion (DD) method of antifungal susceptibility (AFS) was evaluated for three different classes of antifungals: amphotericin B (AMB), voriconazole (VCZ) and caspofungin (CAS). Forty four clinical isolates of Aspergillus were selected; these included 34 A.fumigatus, 8 A.flavus and 2 A. terreus. AFS by DD and E-test was done on non-supplemented Mueller Hinton Agar (MHA) and was compared to Clinical Laboratory Standard Institute(CLSI) broth micro-dilution (BMD) method of AFS. Results: Disk diffusion method for amphotericin B showed 87.5% agreement while E-test showed 93.8% agreement with broth micro-dilution. The agreement with broth micro-dilution was similar for both disk diffusion and E-test in case of voriconazole (93.8%) and caspofungin (100%). 31.8% and 9.1% Aspergillus isolates were found to have amphotericin B and voriconazole MIC values above epidemiological cut off value (ECV) respectively. All isolates were within ECV for caspofungin. Conclusion: CLSI method of DD promises to be easier, reproducible and cost effective method of susceptibility testing, but this method must be interpreted with caution in case of amphotericin B susceptibility testing. E-test correlates better than DD with BMD. PMID:25737984

Khare, Vineeta; Kumar, Deepak; Ahmad, Abrar; Banerjee, Gopa; Singh, Mastan

2015-01-01

425

Genetic Similarity among One Aspergillus flavus Strain Isolated from a Patient Who Underwent Heart Surgery and Two Environmental Strains Obtained from the Operating Room  

PubMed Central

We report the simultaneous isolation of one Aspergillus flavus strain from the aortic prosthesis of a heart surgery patient and another two isolates recovered from a dual-reservoir cooler-heater used in the operating room where this patient was operated on. Genetic typing of these three isolates by randomly amplified polymorphic DNA (RAPD) revealed identical genotypes. Eight unrelated control strains of A. flavus had eight different genotypes. These results clearly indicated the nosocomial origin of the A. flavus strain isolated from the patient. We suggest that the RAPD technique is a rapid and reliable tool to ascertain the epidemiology of infections caused by A. flavus. PMID:10835021

Diaz-Guerra, Teresa M.; Mellado, Emilia; Cuenca-Estrella, Manuel; Gaztelurrutia, Lourdes; Navarro, Jose Ignacio Villate; Tudela, Juan L. Rodríguez

2000-01-01

426