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Sample records for der hefe yarrowia

  1. Yarrowia van der Walt & von Arx (1980)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This book chapter describes the ascomycete yeast genus Yarrowia and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus has just one described species, Y. lipolytica, and is commonly known by its asexual name Candida lipolytica. The species is widely distributed in name and...

  2. Fungemia caused by Yarrowia lipolytica.

    PubMed

    Trabelsi, H; Chtara, K; Khemakhem, N; Néji, S; Cheikhrouhou, F; Sellami, H; Guidara, R; Makni, F; Bouaziz, M; Ayadi, A

    2015-06-01

    Yarrowia lipolytica is weakly pathogenic yeast, which is rarely isolated from the blood. We report unusual cases of Y. lipolytica fungemia occurred between October 2012 and June 2014 in the intensive care unit (ICU) of the UH Habib Bourguiba Sfax. During this period, 55 cases of Y. lipolytica septicemia were diagnosed. There were 44 men and 11 women (sex ratio = 4).The median age was 43 years. The broad-spectrum antibiotics (100 %), the catheterization (96 %), and the prolonged hospitalization in ICU (91 %) were the main risk factors. Patients were hospitalized in ICU, mostly, for polytraumatism (45.4 %), pneumopathy (9 %), and post-operative complications (7 %). Fever unresponsive to broad-spectrum antibacterial therapy was the predominant sign of infection (83.6 %). Y. lipolytica was isolated in one or several blood cultures (14.5 %) and in the catheter tip culture of nine patients (16.3 %).Treatment was based on intravenous amphotericin B (58.2 %), fluconazole (45.4 %) and/or removal catheter (69 %). Apyrexia or blood cultures sterilization was obtained for 34 patients (61.8 %). Y. lipolytica candidemia is an opportunistic and emerging human yeast pathogen. It can reach to the bloodstream of immunocompromised or critically ill patients during hospitalization through intravascular catheterization. Further clinical data need to be evaluated for formulating management strategies of seriously ill patients infected with uncommon fungal agents. PMID:25614084

  3. [Antistress systems of the yeast Yarrowia lipolitica (review)].

    PubMed

    Arinbasarova, A Yu; Biryukova, E N; Medentsev, A G

    2015-01-01

    The authors' and literature data on the adaptation response of the micromycetes Yarrowia lipolytica to various stress impacts are considered in the review. The uniformity of cellular response to all stress factors is discussed. PMID:26027347

  4. Engineering Yarrowia lipolytica for Campesterol Overproduction.

    PubMed

    Du, Hao-Xing; Xiao, Wen-Hai; Wang, Ying; Zhou, Xiao; Zhang, Yu; Liu, Duo; Yuan, Ying-Jin

    2016-01-01

    Campesterol is an important precursor for many sterol drugs, e.g. progesterone and hydrocortisone. In order to produce campesterol in Yarrowia lipolytica, C-22 desaturase encoding gene ERG5 was disrupted and the heterologous 7-dehydrocholesterol reductase (DHCR7) encoding gene was constitutively expressed. The codon-optimized DHCR7 from Rallus norvegicus, Oryza saliva and Xenapus laevis were explored and the strain with the gene DHCR7 from X. laevis achieved the highest titer of campesterol due to D409 in substrate binding sites. In presence of glucose as the carbon source, higher biomass conversion yield and product yield were achieved in shake flask compared to that using glycerol and sunflower seed oil. Nevertheless, better cell growth rate was observed in medium with sunflower seed oil as the sole carbon source. Through high cell density fed-batch fermentation under carbon source restriction strategy, a titer of 453±24.7 mg/L campesterol was achieved with sunflower seed oil as the carbon source, which is the highest reported microbial titer known. Our study has greatly enhanced campesterol accumulation in Y. lipolytica, providing new insight into producing complex and desired molecules in microbes. PMID:26751680

  5. Engineering Yarrowia lipolytica for Campesterol Overproduction

    PubMed Central

    Zhang, Yu; Liu, Duo; Yuan, Ying-Jin

    2016-01-01

    Campesterol is an important precursor for many sterol drugs, e.g. progesterone and hydrocortisone. In order to produce campesterol in Yarrowia lipolytica, C-22 desaturase encoding gene ERG5 was disrupted and the heterologous 7-dehydrocholesterol reductase (DHCR7) encoding gene was constitutively expressed. The codon-optimized DHCR7 from Rallus norvegicus, Oryza saliva and Xenapus laevis were explored and the strain with the gene DHCR7 from X. laevis achieved the highest titer of campesterol due to D409 in substrate binding sites. In presence of glucose as the carbon source, higher biomass conversion yield and product yield were achieved in shake flask compared to that using glycerol and sunflower seed oil. Nevertheless, better cell growth rate was observed in medium with sunflower seed oil as the sole carbon source. Through high cell density fed-batch fermentation under carbon source restriction strategy, a titer of 453±24.7 mg/L campesterol was achieved with sunflower seed oil as the carbon source, which is the highest reported microbial titer known. Our study has greatly enhanced campesterol accumulation in Y. lipolytica, providing new insight into producing complex and desired molecules in microbes. PMID:26751680

  6. Tunable nano-oleosomes derived from engineered Yarrowia lipolytica.

    PubMed

    Han, Zhenlin; Madzak, Catherine; Su, Wei Wen

    2013-03-01

    Oleosomes are discrete organelles filled with neutral lipids surrounded by a protein-embedded phospholipid monolayer. Their simple yet robust structure, as well as their amenability to biological, chemical, and physical processing, can be exploited for various biotechnology applications. In this study, we report facile biosynthesis of functionalized oleosomes within oleaginous yeast Yarrowia lipolytica, through expression of oleosin fusion proteins. By fusing a cDNA clone of a sesame oleosin with either the coding sequence of a red fluorescent protein mCherry or a cellulosomal scaffolding protein cohesin from Clostridium cellulolyticum, these oleosin-fusion proteins were efficiently expressed and specifically targeted to and anchored on the surface of the oleosomes within the Y. lipolytica cells. The engineered oleosomes can be easily separated from the Y. lipolytica cell extract via floating centrifugation and both mCherry and cohesin domains are shown to be functional. Upon sonication, the engineered Yarrowia oleosomes exhibit a mean diameter of 200-300 nm and are found to be highly stable. The feasibility of co-displaying multiple proteins on the Yarrowia oleosomes was demonstrated by incubating cohesin-displaying oleosomes with different dockerin-fusion proteins. Based on this strategy, engineered oleosomes with both cell-targeting and reporting activities were created and shown to be functional. Taken together, the Yarrowia oleosome surface display system in which oleosin serves as an efficient membrane anchor motif shows great promise as a simple platform for creating tunable nanoparticles. PMID:23096765

  7. Yarrowia lipolytica and pollutants: Interactions and applications.

    PubMed

    Zinjarde, Smita; Apte, Mugdha; Mohite, Pallavi; Kumar, Ameeta Ravi

    2014-01-01

    Yarrowia lipolytica is a dimorphic, non-pathogenic, ascomycetous yeast species with distinctive physiological features and biochemical characteristics that are significant in environment-related matters. Strains naturally present in soils, sea water, sediments and waste waters have inherent abilities to degrade hydrocarbons such as alkanes (short and medium chain) and aromatic compounds (biphenyl and dibenzofuran). With the application of slow release fertilizers, design of immobilization techniques and development of microbial consortia, scale-up studies and in situ applications have been possible. In general, hydrocarbon uptake in this yeast is mediated by attachment to large droplets (via hydrophobic cell surfaces) or is aided by surfactants and emulsifiers. Subsequently, the internalized hydrocarbons are degraded by relevant enzymes innately present in the yeast. Some wild-type or recombinant strains also detoxify nitroaromatic (2,4,6-trinitrotoluene), halogenated (chlorinated and brominated hydrocarbons) and organophosphate (methyl parathion) compounds. The yeast can tolerate some metals and detoxify them via different biomolecules. The biomass (unmodified, in combination with sludge, magnetically-modified and in the biofilm form) has been employed in the biosorption of hexavalent chromium ions from aqueous solutions. Yeast cells have also been applied in protocols related to nanoparticle synthesis. The treatment of oily and solid wastes with this yeast reduces chemical oxygen demand or value-added products (single cell oil, single cell protein, surfactants, organic acids and polyalcohols) are obtained. On account of all these features, the microorganism has established a place for itself and is of considerable value in environment-related applications. PMID:24780156

  8. Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica.

    PubMed

    Wang, Wei; Wei, Hui; Alahuhta, Markus; Chen, Xiaowen; Hyman, Deborah; Johnson, David K; Zhang, Min; Himmel, Michael E

    2014-01-01

    To develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. The successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism. PMID:25462572

  9. Heterologous Expression of Xylanase Enzymes in Lipogenic Yeast Yarrowia lipolytica

    PubMed Central

    Alahuhta, Markus; Chen, Xiaowen; Hyman, Deborah; Johnson, David K.; Zhang, Min; Himmel, Michael E.

    2014-01-01

    To develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. The successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism. PMID:25462572

  10. Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica

    DOE PAGESBeta

    Wang, Wei; Wei, Hui; Alahuhta, Markus; Chen, Xiaowen; Hyman, Deborah; Johnson, David K.; Zhang, Min; Himmel, Michael E.

    2014-12-02

    In order to develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an abilitymore » to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. Finally, the successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism.« less

  11. Enantiocomplementary Yarrowia lipolytica Oxidoreductases: Alcohol Dehydrogenase 2 and Short Chain Dehydrogenase/Reductase

    PubMed Central

    Napora-Wijata, Kamila; Strohmeier, Gernot A.; Sonavane, Manoj N.; Avi, Manuela; Robins, Karen; Winkler, Margit

    2013-01-01

    Enzymes of the non-conventional yeast Yarrowia lipolytica seem to be tailor-made for the conversion of lipophilic substrates. Herein, we cloned and overexpressed the Zn-dependent alcohol dehydrogenase ADH2 from Yarrowia lipolytica in Escherichia coli. The purified enzyme was characterized in vitro. The substrate scope for YlADH2 mediated oxidation and reduction was investigated spectrophotometrically and the enzyme showed a broader substrate range than its homolog from Saccharomyces cerevisiae. A preference for secondary compared to primary alcohols in oxidation direction was observed for YlADH2. 2-Octanone was investigated in reduction mode in detail. Remarkably, YlADH2 displays perfect (S)-selectivity and together with a highly (R)-selective short chain dehydrogenase/ reductase from Yarrowia lipolytica it is possible to access both enantiomers of 2-octanol in >99% ee with Yarrowia lipolytica oxidoreductases. PMID:24970175

  12. Metabolic engineering of Yarrowia lipolytica for industrial applications.

    PubMed

    Zhu, Quinn; Jackson, Ethel N

    2015-12-01

    Yarrowia lipolytica is a safe and robust yeast that has a history of industrial applications. Its physiological, metabolic and genomic characteristics have made it a superior host for metabolic engineering. The results of optimizing internal pathways and introducing new pathways have demonstrated that Y. lipolytica can be a platform cell factory for cost-effective production of chemicals and fuels derived from fatty acids, lipids and acetyl-CoA. Two products have been commercialized from metabolically engineered Y. lipolytica strains producing high amounts of omega-3 eicosapentaenoic acid, and more products are on the way to be produced at industrial scale. Here we review recent progress in metabolic engineering of Y. lipolytica for production of biodiesel fuel, functional fatty acids and carotenoids. PMID:26319895

  13. Synthesis of biodegradable polymers using biocatalysis with Yarrowia lipolytica lipase.

    PubMed

    Barrera-Rivera, Karla A; Flores-Carreón, Arturo; Martínez-Richa, Antonio

    2012-01-01

    Yarrowia lipolytica lipase (YLL) was used as catalyst in the enzymatic ring-opening polymerization (ROP) of ε-caprolactone. This low-cost solid-state lipase produces low-molecular-weight polyesters with unique multiphase morphology as determined by carbon-13 NMR. YLL attaches sugar head groups to polycaprolactone in a one-pot biocatalytic pathway. Synthesis of α-ω-telechelic (polymer with two reactive hydroxyl end groups) PCL diols is achieved by enzymatic ROP with YLL immobilized on the macroporous resin Lewatit VPOC 1026, and in the presence of diethylene glycol or poly(ethylene glycol). Biodegradable linear polyester urethanes are prepared by polycondensation between synthesized PCL diols and hexamethylene-diisocyanate. PMID:22426736

  14. Biotechnological applications of Yarrowia lipolytica: Past, present and future.

    PubMed

    Liu, Hu-Hu; Ji, Xiao-Jun; Huang, He

    2015-12-01

    Non-conventional yeasts have attracted increasing interest due to their biochemical characteristics and potential applications. Yarrowia lipolytica is a non-conventional yeast with specific characteristics and physiology. The potential physiological and metabolic capabilities of Y. lipolytica, which can assimilate many different carbon sources, including typical hydrophilic and hydrophobic materials, are reviewed in this paper. Concerning the uptake and metabolism substrates, this review focuses particularly on low-cost raw materials, such as glycerol. Moreover, this review presents the results of safety studies of Y. lipolytica. Finally, the wide applications of Y. lipolytica, such as functional enzyme production, metabolite synthesis and environmental bioremediation, are reviewed in this paper. Recently, with the development of system biology and synthetic biology, it was concluded that these technologies will provide new opportunities for potential applications of Y. lipolytica in the future. PMID:26248319

  15. Identification and application of keto acids transporters in Yarrowia lipolytica

    PubMed Central

    Guo, Hongwei; Liu, Peiran; Madzak, Catherine; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2015-01-01

    Production of organic acids by microorganisms is of great importance for obtaining building-block chemicals from sustainable biomass. Extracellular accumulation of organic acids involved a series of transporters, which play important roles in the accumulation of specific organic acid while lack of systematic demonstration in eukaryotic microorganisms. To circumvent accumulation of by-product, efforts have being orchestrated to carboxylate transport mechanism for potential clue in Yarrowia lipolytica WSH-Z06. Six endogenous putative transporter genes, YALI0B19470g, YALI0C15488g, YALI0C21406g, YALI0D24607g, YALI0D20108g and YALI0E32901g, were identified. Transport characteristics and substrate specificities were further investigated using a carboxylate-transport-deficient Saccharomyces cerevisiae strain. These transporters were expressed in Y. lipolytica WSH-Z06 to assess their roles in regulating extracellular keto acids accumulation. In a Y. lipolytica T1 line over expressing YALI0B19470g, α-ketoglutarate accumulated to 46.7 g·L−1, whereas the concentration of pyruvate decreased to 12.3 g·L−1. Systematic identification of these keto acids transporters would provide clues to further improve the accumulation of specific organic acids with higher efficiency in eukaryotic microorganisms. PMID:25633653

  16. Metabolic engineering of Yarrowia lipolytica for itaconic acid production.

    PubMed

    Blazeck, John; Hill, Andrew; Jamoussi, Mariam; Pan, Anny; Miller, Jarrett; Alper, Hal S

    2015-11-01

    Itaconic acid is a naturally produced organic acid with diverse applications as a replacement for petroleum derived products. However, its industrial viability as a bio-replacement has been restricted due to limitations with native producers. In this light, Yarrowia lipolytica is an excellent potential candidate for itaconic acid production due to its innate capacity to accumulate citric acid cycle intermediates and tolerance to lower pH. Here, we demonstrate the capacity to produce itaconic acid in Y. lipolytica through heterologous expression of the itaconic acid synthesis enzyme, resulting in an initial titer of 33 mg/L. Further optimizations of this strain via metabolic pathway engineering, enzyme localization, and media optimization strategies enabled 4.6g/L of itaconic acid to be produced in bioreactors, representing a 140-fold improvement over initial titer. Moreover, these fermentation conditions did not require additional nutrient supplementation and utilized a low pH condition that enabled the acid form of itaconic acid to be produced. Overall yields (0.058 g/g yield from glucose) and maximum productivity of 0.045 g/L/h still provide areas for future strain improvement. Nevertheless, this work demonstrates that Y. lipolytica has the potential to serve as an industrially relevant platform for itaconic acid production. PMID:26384571

  17. Engineering lipid overproduction in the oleaginous yeast Yarrowia lipolytica.

    PubMed

    Qiao, Kangjian; Imam Abidi, Syed Hussain; Liu, Hongjuan; Zhang, Haoran; Chakraborty, Sagar; Watson, Nicki; Kumaran Ajikumar, Parayil; Stephanopoulos, Gregory

    2015-05-01

    Conversion of carbohydrates to lipids at high yield and productivity is essential for cost-effective production of renewable biodiesel. Although some microorganisms can convert sugars to oils, conversion yields and rates are typically low due primarily to allosteric inhibition of the lipid biosynthetic pathway by saturated fatty acids. By reverse engineering the mammalian cellular obese phenotypes, we identified the delta-9 stearoyl-CoA desaturase (SCD) as a rate limiting step and target for the metabolic engineering of the lipid synthesis pathway in Yarrowia lipolytica. Simultaneous overexpression of SCD, Acetyl-CoA carboxylase (ACC1), and Diacylglyceride acyl-transferase (DGA1) in Y. lipolytica yielded an engineered strain exhibiting highly desirable phenotypes of fast cell growth and lipid overproduction including high carbon to lipid conversion yield (84.7% of theoretical maximal yield), high lipid titers (~55g/L), enhanced tolerance to glucose and cellulose-derived sugars. Moreover, the engineered strain featured a three-fold growth advantage over the wild type strain. As a result, a maximal lipid productivity of ~1g/L/h is obtained during the stationary phase. Furthermore, we showed that the engineered yeast required cytoskeleton remodeling in eliciting the obesity phenotype. Altogether, our work describes the development of a microbial catalyst with the highest reported lipid yield, titer and productivity to date. This is an important step towards the development of an efficient and cost-effective process for biodiesel production from renewable resources. PMID:25732624

  18. Identification and application of keto acids transporters in Yarrowia lipolytica.

    PubMed

    Guo, Hongwei; Liu, Peiran; Madzak, Catherine; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2015-01-01

    Production of organic acids by microorganisms is of great importance for obtaining building-block chemicals from sustainable biomass. Extracellular accumulation of organic acids involved a series of transporters, which play important roles in the accumulation of specific organic acid while lack of systematic demonstration in eukaryotic microorganisms. To circumvent accumulation of by-product, efforts have being orchestrated to carboxylate transport mechanism for potential clue in Yarrowia lipolytica WSH-Z06. Six endogenous putative transporter genes, YALI0B19470g, YALI0C15488g, YALI0C21406g, YALI0D24607g, YALI0D20108g and YALI0E32901g, were identified. Transport characteristics and substrate specificities were further investigated using a carboxylate-transport-deficient Saccharomyces cerevisiae strain. These transporters were expressed in Y. lipolytica WSH-Z06 to assess their roles in regulating extracellular keto acids accumulation. In a Y. lipolytica T1 line over expressing YALI0B19470g, α-ketoglutarate accumulated to 46.7 g·L(-1), whereas the concentration of pyruvate decreased to 12.3 g·L(-1). Systematic identification of these keto acids transporters would provide clues to further improve the accumulation of specific organic acids with higher efficiency in eukaryotic microorganisms. PMID:25633653

  19. Yarrowia lipolytica and its multiple applications in the biotechnological industry.

    PubMed

    Gonçalves, F A G; Colen, G; Takahashi, J A

    2014-01-01

    Yarrowia lipolytica is a nonpathogenic dimorphic aerobic yeast that stands out due to its ability to grow in hydrophobic environments. This property allowed this yeast to develop an ability to metabolize triglycerides and fatty acids as carbon sources. This feature enables using this species in the bioremediation of environments contaminated with oil spill. In addition, Y. lipolytica has been calling the interest of researchers due to its huge biotechnological potential, associated with the production of several types of metabolites, such as bio-surfactants, γ-decalactone, citric acid, and intracellular lipids and lipase. The production of a metabolite rather than another is influenced by the growing conditions to which Y. lipolytica is subjected. The choice of carbon and nitrogen sources to be used, as well as their concentrations in the growth medium, and the careful determination of fermentation parameters, pH, temperature, and agitation (oxygenation), are essential for efficient metabolites production. This review discusses the biotechnological potential of Y. lipolytica and the best growing conditions for production of some metabolites of biotechnological interest. PMID:24715814

  20. Yarrowia lipolytica and Its Multiple Applications in the Biotechnological Industry

    PubMed Central

    Gonçalves, F. A. G.; Colen, G.; Takahashi, J. A.

    2014-01-01

    Yarrowia lipolytica is a nonpathogenic dimorphic aerobic yeast that stands out due to its ability to grow in hydrophobic environments. This property allowed this yeast to develop an ability to metabolize triglycerides and fatty acids as carbon sources. This feature enables using this species in the bioremediation of environments contaminated with oil spill. In addition, Y. lipolytica has been calling the interest of researchers due to its huge biotechnological potential, associated with the production of several types of metabolites, such as bio-surfactants, γ-decalactone, citric acid, and intracellular lipids and lipase. The production of a metabolite rather than another is influenced by the growing conditions to which Y. lipolytica is subjected. The choice of carbon and nitrogen sources to be used, as well as their concentrations in the growth medium, and the careful determination of fermentation parameters, pH, temperature, and agitation (oxygenation), are essential for efficient metabolites production. This review discusses the biotechnological potential of Y. lipolytica and the best growing conditions for production of some metabolites of biotechnological interest. PMID:24715814

  1. Applications of the Non-Conventional Yeast Yarrowia lipolytica

    NASA Astrophysics Data System (ADS)

    Thevenieau, France; Nicaud, Jean-Marc; Gaillardin, Claude

    The yeast Yarrowia lipolytica is often found associated to proteinaceous or hydrophobic substrates such as alkanes or lipids. To assimilate these hydropho-bic substrates, Y. lipolytica has developed an adaptative strategy resulting in elaborated morphological and physiological changes leading to terminal and β-oxidation of substrates as well as to lipid storage. The completion of the Y. lipolytica genome greatly improved our understanding of these mechanisms. Three main applications of this metabolism will be discussed. The first class corresponds to bioconver-sion processes for the production of secondary metabolites (citric acid), of aroma ( γ - lactone, green note, epoxy geraniol) and of chemicals (dicarboxylic acids). The second class leads to fine chemical production by enantio separation of pharmaceutical compounds using Y. lipolytica enzymes such as epoxyde hydrolase or lipase. The third one refers to production of Single Cell Oils (SCO) from agriculture feedstock. In addition to its ability to handle hydrophobic substrates, Y. lipolytica has also been recognised as a strong secretor of various proteins such as proteases, lipases, RNases and others. A comprehensive review of recent developments of the Y. lipolytica expression/secretion system will finally be presented.

  2. Factorial Design to Optimize Biosurfactant Production by Yarrowia lipolytica

    PubMed Central

    Fontes, Gizele Cardoso; Fonseca Amaral, Priscilla Filomena; Nele, Marcio; Zarur Coelho, Maria Alice

    2010-01-01

    In order to improve biosurfactant production by Yarrowia lipolytica IMUFRJ 50682, a factorial design was carried out. A 24 full factorial design was used to investigate the effects of nitrogen sources (urea, ammonium sulfate, yeast extract, and peptone) on maximum variation of surface tension (ΔST) and emulsification index (EI). The best results (67.7% of EI and 20.9 mN m−1 of ΔST) were obtained in a medium composed of 10 g 1−1 of ammonium sulfate and 0.5 g 1−1 of yeast extract. Then, the effects of carbon sources (glycerol, hexadecane, olive oil, and glucose) were evaluated. The most favorable medium for biosurfactant production was composed of both glucose (4% w/v) and glycerol (2% w/v), which provided an EI of 81.3% and a ΔST of 19.5 mN m−1. The experimental design optimization enhanced ΔEI by 110.7% and ΔST by 108.1% in relation to the standard process. PMID:20368788

  3. Engineering Promoter Architecture in Oleaginous Yeast Yarrowia lipolytica.

    PubMed

    Shabbir Hussain, Murtaza; Gambill, Lauren; Smith, Spencer; Blenner, Mark A

    2016-03-18

    Eukaryotic promoters have a complex architecture to control both the strength and timing of gene transcription spanning up to thousands of bases from the initiation site. This complexity makes rational fine-tuning of promoters in fungi difficult to predict; however, this very same complexity enables multiple possible strategies for engineering promoter strength. Here, we studied promoter architecture in the oleaginous yeast, Yarrowia lipolytica. While recent studies have focused on upstream activating sequences, we systematically examined various components common in fungal promoters. Here, we examine several promoter components including upstream activating sequences, proximal promoter sequences, core promoters, and the TATA box in autonomously replicating expression plasmids and integrated into the genome. Our findings show that promoter strength can be fine-tuned through the engineering of the TATA box sequence, core promoter, and upstream activating sequences. Additionally, we identified a previously unreported oleic acid responsive transcription enhancement in the XPR2 upstream activating sequences, which illustrates the complexity of fungal promoters. The promoters engineered here provide new genetic tools for metabolic engineering in Y. lipolytica and provide promoter engineering strategies that may be useful in engineering other non-model fungal systems. PMID:26635071

  4. Disruption of Yarrowia lipolytica biofilms by rhamnolipid biosurfactant

    PubMed Central

    2012-01-01

    Background Yarrowia lipolytica is an ascomycetous dimorphic fungus that exhibits biofilm mode of growth. Earlier work has shown that biosurfactants such as rhamnolipids are efficient dispersants of bacterial biofilms. However, their effectiveness against fungal biofilms (particularly Y. lipolytica) has not been investigated. The aim of this study was to determine the effect of rhamnolipid on a biofilm forming strain of Y. lipolytica. Two chemical surfactants, cetyl-trimethyl ammonium bromide (CTAB) and sodium dodecyl sulphate (SDS) were used as controls for comparison. Results The methylene blue dye exclusion assay indicated an increase in fungal cell permeability after rhamnolipid treatment. Microtiter plate assay showed that the surfactant coating decreased Y. lipolytica biofilm formation by 50%. Rhamnolipid treatment disrupted pre-formed biofilms in a more effective manner than the other two surfactants. Confocal laser scanning microscopic studies showed that biofilm formation onto glass surfaces was decreased by 67% after sub-minimum inhibitory concentration (sub-MIC) treatment with rhamnolipids. The disruption of biofilms after rhamnolipid treatment was significant (P<0.05) when compared to SDS and CTAB. Conclusion The results indicate a potential application of the biological surfactant to disrupt Y. lipolytica biofilms. PMID:22839701

  5. Autocloning and Amplification of LIP2 in Yarrowia lipolytica

    PubMed Central

    Pignède, Georges; Wang, Hui-Jie; Fudalej, Franck; Seman, Michel; Gaillardin, Claude; Nicaud, Jean-Marc

    2000-01-01

    We synthesized a Yarrowia lipolytica strain overproducing lipase for industrial applications by using long terminal repeat (ζ) of the Y. lipolytica retrotransposon Ylt1 and an allele of URA3 with a promoter deletion to construct JMP3. JMP3 is a derivative of plasmid pHSS6 carrying a NotI-NotI cassette which contains a defective URA3 allele, a polylinker sequence, and the ζ region for targeting to multiple sites in the genome of the recipient. We inserted the LIP2 gene (encoding extracellular lipase) under the control of the strong POX2 promoter into JMP3 to generate JMP6. The pHSS6 region was removed by NotI digestion prior to transformation. Two Y. lipolytica strains transformed with the JMP6 LIP2 cassette had a mean of 10 integrated copies devoid of the Escherichia coli region, corresponding to an autocloning event. The copy number in the transformants was stable even after 120 generations in nonselective and lipase-inducing conditions. The resulting strains could produce 0.5 g of active lipase per liter in the supernatant, 40 times more than the single-copy strain with the LIP2 promoter. This work provides a new expression system in Y. lipolytica that results in strains devoid of bacterial DNA and in strains producing a high level of lipase for industrial uses, waste treatment, and pancreatic insufficiency therapy. PMID:10919782

  6. Draft Genome Sequence of the Dimorphic Yeast Yarrowia lipolytica Strain W29

    PubMed Central

    Pomraning, Kyle R.

    2015-01-01

    Here, we present the draft genome sequence of the dimorphic ascomycete yeast Yarrowia lipolytica strain W29 (ATCC 20460). Y. lipolytica is a commonly employed model for the industrial production of lipases, small molecules, and more recently for its ability to accumulate lipids. PMID:26607882

  7. Draft Genome Sequence of the Dimorphic Yeast Yarrowia lipolytica Strain W29.

    PubMed

    Pomraning, Kyle R; Baker, Scott E

    2015-01-01

    Here, we present the draft genome sequence of the dimorphic ascomycete yeast Yarrowia lipolytica strain W29 (ATCC 20460). Y. lipolytica is a commonly employed model for the industrial production of lipases, small molecules, and more recently for its ability to accumulate lipids. PMID:26607882

  8. Comparative physiology of forty-five Yarrowia lipolytica strains grown on pretreated switchgrass hydrolysate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yarrowia lipolytica is a well-characterized yeast of the phylum Ascomycota with established use in the biotechnology industry for production of organic acids and enzymes. In addition, the yeast is a model oleaginous organism that accumulates lipids during growth on a variety of carbon sources. The a...

  9. Protein expression and secretion in the yeast Yarrowia lipolytica.

    PubMed

    Nicaud, Jean-Marc; Madzak, Catherine; van den Broek, Peter; Gysler, Christof; Duboc, Philippe; Niederberger, Peter; Gaillardin, Claude

    2002-08-01

    Strains and vectors for protein expression and secretion have been developed in the yeast Yarrowia lipolytica. Host strains were constructed with non-reverting auxotrophic markers, deletions of protease-encoding genes, and carrying a docking platform. To drive transcription, either the synthetic hp4d or the inducible POX2 promoter were used. Protein secretion is either directed by the targeting sequence of the alkaline extracellular protease or the extracellular lipase (LIP2p) signal sequence. We describe a set of vectors based on these promoters, targeting sequences and two URA3 alleles as selection markers. The wild-type URA3 allele, ura3d1, was used for single-copy integration and a mutant URA3 allele, ura3d4, was used to select for multi-copy integration into the genome. These vectors were used to express the Y. lipolytica extracellular lipase LIP2p and the Aspergillus oryzae leucine amino peptidase II. Lipase production under the control of the hp4d promoter by a strain containing a single copy reached 1000 U ml(-1) in shake flasks, while a strain containing multiple integrations reached 2000 U ml(-1) in shake flasks, 11500 U ml(-1) in batch and 90500 U ml(-1) in fed batch. Leucine amino peptidase production under the control of the hp4d promoter reached 320 mU ml(-1) in batch with a mono-copy lapA integrant and 28000 mU ml(-1) in fed batch with a multi-copy transformant. PMID:12702287

  10. Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica.

    PubMed

    Ryu, Seunghyun; Hipp, Julie; Trinh, Cong T

    2016-02-01

    The oleaginous yeast Yarrowia lipolytica is an industrially important host for production of organic acids, oleochemicals, lipids, and proteins with broad biotechnological applications. Albeit known for decades, the unique native metabolism of Y. lipolytica for using complex fermentable sugars, which are abundant in lignocellulosic biomass, is poorly understood. In this study, we activated and elucidated the native sugar metabolism in Y. lipolytica for cell growth on xylose and cellobiose as well as their mixtures with glucose through comprehensive metabolic and transcriptomic analyses. We identified 7 putative glucose-specific transporters, 16 putative xylose-specific transporters, and 4 putative cellobiose-specific transporters that are transcriptionally upregulated for growth on respective single sugars. Y. lipolytica is capable of using xylose as a carbon source, but xylose dehydrogenase is the key bottleneck of xylose assimilation and is transcriptionally repressed by glucose. Y. lipolytica has a set of 5 extracellular and 6 intracellular β-glucosidases and is capable of assimilating cellobiose via extra- and intracellular mechanisms, the latter being dominant for growth on cellobiose as a sole carbon source. Strikingly, Y. lipolytica exhibited enhanced sugar utilization for growth in mixed sugars, with strong carbon catabolite activation for growth on the mixture of xylose and cellobiose and with mild carbon catabolite repression of glucose on xylose and cellobiose. The results of this study shed light on fundamental understanding of the complex native sugar metabolism of Y. lipolytica and will help guide inverse metabolic engineering of Y. lipolytica for enhanced conversion of biomass-derived fermentable sugars to chemicals and fuels. PMID:26682853

  11. Unraveling fatty acid transport and activation mechanisms in Yarrowia lipolytica.

    PubMed

    Dulermo, Rémi; Gamboa-Meléndez, Heber; Ledesma-Amaro, Rodrigo; Thévenieau, France; Nicaud, Jean-Marc

    2015-09-01

    Fatty acid (FA) transport and activation have been extensively studied in the model yeast species Saccharomyces cerevisiae but have rarely been examined in oleaginous yeasts, such as Yarrowia lipolytica. Because the latter begins to be used in biodiesel production, understanding its FA transport and activation mechanisms is essential. We found that Y. lipolytica has FA transport and activation proteins similar to those of S. cerevisiae (Faa1p, Pxa1p, Pxa2p, Ant1p) but mechanism of FA peroxisomal transport and activation differs greatly with that of S. cerevisiae. While the ScPxa1p/ScPxa2p heterodimer is essential for growth on long-chain FAs, ΔYlpxa1 ΔYlpxa2 is not impaired for growth on FAs. Meanwhile, ScAnt1p and YlAnt1p are both essential for yeast growth on medium-chain FAs, suggesting they function similarly. Interestingly, we found that the ΔYlpxa1 ΔYlpxa2 ΔYlant1 mutant was unable to grow on short-, medium-, or long-chain FAs, suggesting that YlPxa1p, YlPxa2p, and YlAnt1p belong to two different FA degradation pathways. We also found that YlFaa1p is involved in FA storage in lipid bodies and that FA remobilization largely depended on YlFat1p, YlPxa1p and YlPxa2p. This study is the first to comprehensively examine FA intracellular transport and activation in oleaginous yeast. PMID:25887939

  12. Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica

    PubMed Central

    Ryu, Seunghyun; Hipp, Julie

    2015-01-01

    The oleaginous yeast Yarrowia lipolytica is an industrially important host for production of organic acids, oleochemicals, lipids, and proteins with broad biotechnological applications. Albeit known for decades, the unique native metabolism of Y. lipolytica for using complex fermentable sugars, which are abundant in lignocellulosic biomass, is poorly understood. In this study, we activated and elucidated the native sugar metabolism in Y. lipolytica for cell growth on xylose and cellobiose as well as their mixtures with glucose through comprehensive metabolic and transcriptomic analyses. We identified 7 putative glucose-specific transporters, 16 putative xylose-specific transporters, and 4 putative cellobiose-specific transporters that are transcriptionally upregulated for growth on respective single sugars. Y. lipolytica is capable of using xylose as a carbon source, but xylose dehydrogenase is the key bottleneck of xylose assimilation and is transcriptionally repressed by glucose. Y. lipolytica has a set of 5 extracellular and 6 intracellular β-glucosidases and is capable of assimilating cellobiose via extra- and intracellular mechanisms, the latter being dominant for growth on cellobiose as a sole carbon source. Strikingly, Y. lipolytica exhibited enhanced sugar utilization for growth in mixed sugars, with strong carbon catabolite activation for growth on the mixture of xylose and cellobiose and with mild carbon catabolite repression of glucose on xylose and cellobiose. The results of this study shed light on fundamental understanding of the complex native sugar metabolism of Y. lipolytica and will help guide inverse metabolic engineering of Y. lipolytica for enhanced conversion of biomass-derived fermentable sugars to chemicals and fuels. PMID:26682853

  13. Yarrowia lipolytica vesicle-mediated protein transport pathways

    PubMed Central

    Swennen, Dominique; Beckerich, Jean-Marie

    2007-01-01

    Background Protein secretion is a universal cellular process involving vesicles which bud and fuse between organelles to bring proteins to their final destination. Vesicle budding is mediated by protein coats; vesicle targeting and fusion depend on Rab GTPase, tethering factors and SNARE complexes. The Génolevures II sequencing project made available entire genome sequences of four hemiascomycetous yeasts, Yarrowia lipolytica, Debaryomyces hansenii, Kluyveromyces lactis and Candida glabrata. Y. lipolytica is a dimorphic yeast and has good capacities to secrete proteins. The translocation of nascent protein through the endoplasmic reticulum membrane was well studied in Y. lipolytica and is largely co-translational as in the mammalian protein secretion pathway. Results We identified S. cerevisiae proteins involved in vesicular secretion and these protein sequences were used for the BLAST searches against Génolevures protein database (Y. lipolytica, C. glabrata, K. lactis and D. hansenii). These proteins are well conserved between these yeasts and Saccharomyces cerevisiae. We note several specificities of Y. lipolytica which may be related to its good protein secretion capacities and to its dimorphic aspect. An expansion of the Y. lipolytica Rab protein family was observed with autoBLAST and the Rab2- and Rab4-related members were identified with BLAST against NCBI protein database. An expansion of this family is also found in filamentous fungi and may reflect the greater complexity of the Y. lipolytica secretion pathway. The Rab4p-related protein may play a role in membrane recycling as rab4 deleted strain shows a modification of colony morphology, dimorphic transition and permeability. Similarly, we find three copies of the gene (SSO) encoding the plasma membrane SNARE protein. Quantification of the percentages of proteins with the greatest homology between S. cerevisiae, Y. lipolytica and animal homologues involved in vesicular transport shows that 40% of Y

  14. Draft genome sequence of the dimorphic yeast Yarrowia lipolytica, strain W29

    SciTech Connect

    Pomraning, Kyle R.; Baker, Scott E.

    2015-11-25

    Here we present the draft genome sequence of the dimorphic ascomycete yeast Yarrowia lipolytica strain W29 (ATCC20460TM). Y. lipolytica is a commonly employed model for industrial production of lipases, small molecules, and more recently for its ability to accumulate lipids. It has also been used to study genome evolution in yeast and filamentous fungi due to its position as an early diverging branch of the subphylum Sacchromycotina.

  15. Isolation of bioactive peptides from tryptone that modulate lipase production in Yarrowia lipolytica.

    PubMed

    Turki, Saoussen; Kraeim, Ines Ben; Weeckers, Frederic; Thonart, Philippe; Kallel, Héla

    2009-05-01

    In this work the effect of several organic nitrogen sources on lipase production in Yarrowia lipolytica LgX64.81 overproducing mutant was studied. Among them, tryptone and peptone showed the most prominent stimulatory effect. Interestingly, only tryptic and peptic casein digest were found to highly induce lipase biosynthesis while lipase production was very limited in the presence of casein digest from papain and pronase-catalysed hydrolysis and absent in case of chymotryptic digest. It was also demonstrated that the stimulatory peptides should be present in the culture medium at specific proportions and molecular size to match the physiological requirement of Yarrowia lipolytica strain for lipase biosynthesis. Herein, the lipase-production stimulatory peptides were isolated by ion exchange chromatography for the first time. These results had contributed to gain an insight on tryptone role in lipase production by Yarrowia lipolytica. Moreover the use of a chemically defined medium supplemented with the isolated peptides, will improve the efficiency of the process for lipase production in this yeast. PMID:19188062

  16. Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica

    SciTech Connect

    Wang, Wei; Wei, Hui; Alahuhta, Markus; Chen, Xiaowen; Hyman, Deborah; Johnson, David K.; Zhang, Min; Himmel, Michael E.

    2014-12-02

    In order to develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. Finally, the successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism.

  17. Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering.

    PubMed

    Madzak, Catherine

    2015-06-01

    The oleaginous yeast Yarrowia lipolytica has become a recognized system for expression/secretion of heterologous proteins. This non-conventional yeast is currently being developed as a workhorse for biotechnology by several research groups throughout the world, especially for single-cell oil production, whole cell bioconversion and upgrading of industrial wastes. This mini-review presents established tools for protein expression in Y. lipolytica and highlights novel developments in the areas of promoter design, surface display, and host strain or metabolic pathway engineering. An overview of the industrial and commercial biotechnological applications of Y. lipolytica is also presented. PMID:25947247

  18. Multi-omics analysis reveals regulators of the response to nitrogen limitation in Yarrowia lipolytica

    DOE PAGESBeta

    Pomraning, Kyle R.; Kim, Young -Mo; Nicora, Carrie D.; Chu, Rosalie K.; Bredeweg, Erin L.; Purvine, Samuel O.; Hu, Dehong; Metz, Thomas O.; Baker, Scott E.

    2016-02-25

    Yarrowia lipolytica is an oleaginous ascomycete yeast that stores lipids in response to limitation of nitrogen. Furthermore, while the enzymatic pathways responsible for neutral lipid accumulation in Y. lipolytica are well characterized, regulation of these pathways has received little attention. We therefore sought to characterize the response to nitrogen limitation at system-wide levels, including the proteome, phosphoproteome and metabolome, to better understand how this organism regulates and controls lipid metabolism and to identify targets that may be manipulated to improve lipid yield.

  19. Engineering towards a complete heterologous cellulase secretome in Yarrowia lipolytica reveals its potential for consolidated bioprocessing

    SciTech Connect

    Wei, Hui; Wang, Wei; Alahuhta, Markus; Vander Wall, Todd; Baker, John O.; Taylor, Larry E.; Decker, Stephen R.; Himmel, Michael E.; Zhang, Min

    2014-10-16

    Background: Yarrowia lipolytica is an oleaginous yeast capable of metabolizing glucose to lipids, which then accumulate intracellularly. However, it lacks the suite of cellulolytic enzymes required to break down biomass cellulose and cannot therefore utilize biomass directly as a carbon source. Toward the development of a direct microbial conversion platform for the production of hydrocarbon fuels from cellulosic biomass, the potential for Y. lipolytica to function as a consolidated bioprocessing strain was investigated by first conducting a genomic search and functional testing of its endogenous glycoside hydrolases. Once the range of endogenous enzymes was determined, the critical cellulases from Trichoderma reesei were cloned into Yarrowia. Results: Initially, work to express T. reesei endoglucanase II (EGII) and cellobiohydrolase (CBH) II in Y. lipolytica resulted in the successful secretion of active enzymes. However, a critical cellulase, T. reesei CBHI, while successfully expressed in and secreted from Yarrowia, showed less than expected enzymatic activity, suggesting an incompatibility (probably at the post-translational level) for its expression in Yarrowia. This result prompted us to evaluate alternative or modified CBHI enzymes. Our subsequent expression of a T. reesei-Talaromyces emersonii (Tr-Te) chimeric CBHI, Chaetomium thermophilum CBHI, and Humicola grisea CBHI demonstrated remarkably improved enzymatic activities. Specifically, the purified chimeric Tr-Te CBHI showed a specific activity on Avicel that is comparable to that of the native T. reesei CBHI. Furthermore, the chimeric Tr-Te CBHI also showed significant synergism with EGII and CBHII in degrading cellulosic substrates, using either mixed supernatants or co-cultures of the corresponding Y. lipolytica transformants. The consortia system approach also allows rational volume mixing of the transformant cultures in accordance with the optimal ratio of cellulases required for efficient

  20. Engineering towards a complete heterologous cellulase secretome in Yarrowia lipolytica reveals its potential for consolidated bioprocessing

    DOE PAGESBeta

    Wei, Hui; Wang, Wei; Alahuhta, Markus; Vander Wall, Todd; Baker, John O.; Taylor, Larry E.; Decker, Stephen R.; Himmel, Michael E.; Zhang, Min

    2014-10-16

    Background: Yarrowia lipolytica is an oleaginous yeast capable of metabolizing glucose to lipids, which then accumulate intracellularly. However, it lacks the suite of cellulolytic enzymes required to break down biomass cellulose and cannot therefore utilize biomass directly as a carbon source. Toward the development of a direct microbial conversion platform for the production of hydrocarbon fuels from cellulosic biomass, the potential for Y. lipolytica to function as a consolidated bioprocessing strain was investigated by first conducting a genomic search and functional testing of its endogenous glycoside hydrolases. Once the range of endogenous enzymes was determined, the critical cellulases from Trichodermamore » reesei were cloned into Yarrowia. Results: Initially, work to express T. reesei endoglucanase II (EGII) and cellobiohydrolase (CBH) II in Y. lipolytica resulted in the successful secretion of active enzymes. However, a critical cellulase, T. reesei CBHI, while successfully expressed in and secreted from Yarrowia, showed less than expected enzymatic activity, suggesting an incompatibility (probably at the post-translational level) for its expression in Yarrowia. This result prompted us to evaluate alternative or modified CBHI enzymes. Our subsequent expression of a T. reesei-Talaromyces emersonii (Tr-Te) chimeric CBHI, Chaetomium thermophilum CBHI, and Humicola grisea CBHI demonstrated remarkably improved enzymatic activities. Specifically, the purified chimeric Tr-Te CBHI showed a specific activity on Avicel that is comparable to that of the native T. reesei CBHI. Furthermore, the chimeric Tr-Te CBHI also showed significant synergism with EGII and CBHII in degrading cellulosic substrates, using either mixed supernatants or co-cultures of the corresponding Y. lipolytica transformants. The consortia system approach also allows rational volume mixing of the transformant cultures in accordance with the optimal ratio of cellulases required for efficient

  1. Umsetzung der Unternehmensstrategie mit der Balanced Scorecard

    NASA Astrophysics Data System (ADS)

    Crespo, Isabel; Bergmann, Lars; Portmann, Stefan; Lacker, Thomas; Lacker, Michael; Fleischmann, Jürgen; Kozó, Hans

    Die Balanced Scorecard (BSC) ist ein Ansatz zum strategischen Management, der neben der Ausrichtung des Unternehmens auf finanzielle Zielwerte ebenso großes Gewicht auf so genannte weiche Faktoren legt, die den wirtschaftlichen Erfolg eines Unternehmens erst ermöglichen. Das entscheidende Merkmal der Balanced Scorecard ist dabei, dass sie ein ausgewogenes System strategischer Ziele herstellt, welches das Unternehmen hinsichtlich der vier Perspektiven Finanzen, Kunden, interne Prozesse und Mitarbeiter und Potenziale strategisch ausrichtet (Kaplan u. Norton 1997).

  2. Production of Yarrowia lipolytica Nha2 Na+/H+ antiporter improves the salt tolerance of Saccharomyces cerevisiae.

    PubMed

    Papousková, K; Sychrová, H

    2007-01-01

    Yarrowia lipolytica plasma-membrane Na+/H+ antiporter, encoded by the YlNHA2 gene, is a very efficient exporter of surplus sodium from the cytosol. Its heterologous expression in Saccharomyces cerevisiae wild-type laboratory strains increased their sodium tolerance more efficiently than the expression of ZrSod2-22 antiporter from the osmotolerant yeast Zygosaccharomvces rouxii. PMID:18450222

  3. Expression of a lipid-inducible, self-regulating form of Yarrowia lipolytica lipase LIP2 in Saccharomyces cerevisiae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Yarrowia lipolytica lipase 2 gene (YlLIP2) was cloned into galactose- and fatty acid-inducible Saccharomyces cerevisiae expression vectors and used to generate yeast strains that secrete active LIP2 enzyme activity, as evidenced by results from gene expression analysis and tributyrin turbidity c...

  4. Comprehensive Analysis of a Yeast Lipase Family in the Yarrowia Clade

    PubMed Central

    Devillers, Hugo; Nicaud, Jean-Marc; Marty, Alain; Neuvéglise, Cécile

    2015-01-01

    Lipases are currently the subject of intensive studies due to their large range of industrial applications. The Lip2p lipase from the yeast Yarrowia lipolytica (YlLIP2) was recently shown to be a good candidate for different biotechnological applications. Using a combination of comparative genomics approaches based on sequence similarity, synteny conservation, and phylogeny, we constructed the evolutionary scenario of the lipase family for six species of the Yarrowia clade. RNA-seq based transcriptome analysis revealed the primary role of LIP2 homologues in the assimilation of different substrates. Once identified, these YlLIP2 homologues were expressed in Y. lipolytica. The lipase Lip2a from Candida phangngensis was shown to naturally present better activity and enantioselectivity than YlLip2. Enantioselectivity was further improved by site-directed mutagenesis targeted to the substrate binding site. The mono-substituted variant V232S displayed enantioselectivity greater than 200 and a 2.5 fold increase in velocity. A double-substituted variant 97A-V232F presented reversed enantioselectivity, with a total preference for the R-enantiomer. PMID:26580812

  5. Effect of plant oils upon lipase and citric acid production in Yarrowia lipolytica yeast.

    PubMed

    Darvishi, Farshad; Nahvi, Iraj; Zarkesh-Esfahani, Hamid; Momenbeik, Fariborz

    2009-01-01

    The nonconventional yeast Yarrowia lipolytica degrades very efficiently hydrophobic substrates to produce organic acids, single-cell oil, lipases, and so forth. The aim of this study was to investigate the biochemical behavior and simultaneous production of valuable metabolites such as lipase, citric acid (CA), and single-cell protein (SCP) by Yarrowia lipolytica DSM 3286 grown on various plant oils as sole carbon source. Among tested plant oils, olive oil proved to be the best medium for lipase and CA production. The Y. lipolytica DSM 3286 produced 34.6 +/- 0.1 U/mL of lipase and also CA and SCP as by-product on olive oil medium supplemented with yeast extract. Urea, as organic nitrogen, was the best nitrogen source for CA production. The results of this study suggest that the two biotechnologically valuable products, lipase and CA, could be produced simultaneously by this strain using renewable low-cost substrates such as plant oils in one procedure. PMID:19826636

  6. Effect of Plant Oils upon Lipase and Citric Acid Production in Yarrowia lipolytica Yeast

    PubMed Central

    Darvishi, Farshad; Nahvi, Iraj; Zarkesh-Esfahani, Hamid; Momenbeik, Fariborz

    2009-01-01

    The nonconventional yeast Yarrowia lipolytica degrades very efficiently hydrophobic substrates to produce organic acids, single-cell oil, lipases, and so forth. The aim of this study was to investigate the biochemical behavior and simultaneous production of valuable metabolites such as lipase, citric acid (CA), and single-cell protein (SCP) by Yarrowia lipolytica DSM 3286 grown on various plant oils as sole carbon source. Among tested plant oils, olive oil proved to be the best medium for lipase and CA production. The Y. lipolytica DSM 3286 produced 34.6 ± 0.1 U/mL of lipase and also CA and SCP as by-product on olive oil medium supplemented with yeast extract. Urea, as organic nitrogen, was the best nitrogen source for CA production. The results of this study suggest that the two biotechnologically valuable products, lipase and CA, could be produced simultaneously by this strain using renewable low-cost substrates such as plant oils in one procedure. PMID:19826636

  7. Differential induction, purification and characterization of cold active lipase from Yarrowia lipolytica NCIM 3639.

    PubMed

    Yadav, K N Sathish; Adsul, M G; Bastawde, K B; Jadhav, D D; Thulasiram, H V; Gokhale, D V

    2011-11-01

    The production, purification and characterization of cold active lipases by Yarrowia lipolytica NCIM 3639 is described. The study presents a new finding of production of cell bound and extracellular lipase activities depending upon the substrate used for growth. The strain produced cell bound and extracellular lipase activity when grown on olive oil and Tween 80, respectively. The organism grew profusely at 20 °C and at initial pH of 5.5, producing maximum extracellular lipase. The purified lipase has a molecular mass of 400 kDa having 20 subunits forming a multimeric native protein. Further the enzyme displayed an optimum pH of 5.0 and optimum temperature of 25 °C. Peptide mass finger printing reveled that some peptides showed homologues sequence (42%) to Yarrowia lipolytica LIP8p. The studies on hydrolysis of racemic lavandulyl acetate revealed that extracellular and cell bound lipases show preference over the opposite antipodes of irregular monoterpene, lavandulyl acetate. PMID:21967715

  8. A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

    PubMed Central

    2013-01-01

    Background High supply of raw, residual glycerol from biodiesel production plants promote the search for novel biotechnological methods of its utilization. In this study we attempted modification of glycerol catabolism in a nonconventional yeast species Yarrowia lipolytica through genetic engineering approach. Results To address this, we developed a novel genetic construct which allows transferring three heterologous genes, encoding glycerol dehydratase, its reactivator and a wide-spectrum alcohol oxidoreductase under the control of glycerol-induced promoter. The three genes, tandemly arrayed in an expression cassette with a marker gene ura3, regulatory and targeting sequences (G3P dh promoter and XPR-like terminator, 28S rDNA as a target locus), were transferred into Yarrowia lipolytica cells. The obtained recombinant strain NCYC3825 was characterized at the molecular level and with respect to its biotechnological potential. Our experiments indicated that the novel recombinant strain stably borne one copy of the expression cassette and efficiently expressed heterologous alcohol oxidoreductase, while glycerol dehydratase and its reactivator were expressed at lower level. Comparative shake flask cultivations in glucose- and glycerol-based media demonstrated higher biomass production by the recombinant strain when glycerol was the main carbon source. During bioreactor (5 L) fed-batch cultivation in glycerol-based medium, the recombinant strain was characterized by relatively high biomass and lipids accumulation (up to 42 gDCW L-1, and a peak value of 38%LIPIDS of DCW, respectively), and production of high titers of citric acid (59 g L-1) and 2-phenylethanol (up to 1 g L-1 in shake flask cultivation), which are industrially attractive bioproducts. Conclusions Due to heterogeneous nature of the observed alterations, we postulate that the main driving force of the modified phenotype was faster growth in glycerol-based media, triggered by modifications in the red

  9. Lehrbuch der Mathematischen Physik

    NASA Astrophysics Data System (ADS)

    Thirring, Walter

    Aus den Besprechungen der ersten Auflage: "... Das Buch wendet sich nicht nur an Studierende der Physik und Mathematik in höheren Semestern, sondern ist auch für fertige Physiker und Mathematiker äußerst anregend. Ein besonderes Lob möchte ich für die zahlreichen Illustrationen des Stoffes mit relevanten, anwendungsbezogenen Beispielen aussprechen." Zeitschrift für Angewandte Mathematik und Physik#1 "W. Thirring hat mit seinem Lehrbuch für Mathematische Physik das erstemal den Versuch unternommen, eine spezifische Bearbeitung der gesamten mathematischen Physik einzuleiten und damit ein "Lehrbuch" der wissenschaftlichen Fachwelt vorzulegen, das in seiner Art sicher als neuartig zu bezeichnen ist. Es zeigt neben der Klarheit und Übersichtlichkeit aber auch eine tiefe Kenntnis der Pädagogik, um ein so schwieriges und abstraktes Gebiet darzustellen." Acta Physica Austriaca#2 "Der Leser kann hier Mathematische Physik in einer Form kennenlernen, wie sie sich einem in der akutuellen Forschung stehenden theoretischen Physiker darstellt, der andererseits auch mit der Entwicklung der modernen mathematischen Methoden wohlvertraut ist. Trotz der anspruchsvollen mathematischen Fundierung gerät der Verfasser an keiner Stelle des Buches in die Gefahr, Mathematische Physik im Sinne einer rein mathematischen Disziplin zu verfremden ... Wer heute wissen will, was "Mathematische Physik" ist und sein soll, wird an diesem Buch nicht vorbeigehen können und Antwort und Gewinn davontragen..." ZAMM#3

  10. Lehrbuch der Mathematischen Physik

    NASA Astrophysics Data System (ADS)

    Thirring, Walter

    Aus den Besprechungen der ersten Auflage: "...Das Buch wendet sich nicht nur an Studierende der Physik und Mathematik in höheren Semestern, sondern ist auch für fertige Physiker und Mathematiker äußerst anregend. Ein besonderes Lob möchte ich für die zahlreichen Illustrationen des Stoffes mit relevanten, anwendungsbezogenen Beispielen aussprechen." Zeitschrift für Angewandte Mathematik und Physik#1 "...W. Thirring hat mit seinem Lehrbuch für mathematische Physik das erstemal den Versuch unternommen, eine spezifische Bearbeitung der gesamten mathematischen Physik einzuleiten und damit ein 'Lehrbuch' der wissenschaftlichen Fachwelt vorzulegen, das in seiner Art sicher als neuartig zu bezeichnen ist. Es zeigt neben der Klarheit und Übersichtlichkeit aber auch eine tiefe Kenntnis der Pädagogik, um ein so schwieriges und abstraktes Gebiet darzustellen..." Acta Physica Austriaca#2 "... Der Leser kann hier Mathematische Physik in einer Form kennenlernen, wie sie sich einem in der aktuellen Forschung stehenden theoretischen Physiker darstellt, der andererseits auch mit der Entwicklung der modernen mathematischen Methoden wohlvertraut ist. Trotz der anspruchsvollen mathematischen Fundierung gerät der Verfasser an keiner Stelle des Buches in die Gefahr, Mathematische Physik im Sinne einer rein mathematischen Disziplin zu verfremden...Wer heute wissen will, was 'Mathematische Physik'ist und sein soll, wird an diesem Buch nicht vorbeigehen können und Antwort und Gewinn davontragen..." ZAMM#3

  11. Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production.

    PubMed

    Blazeck, John; Hill, Andrew; Liu, Leqian; Knight, Rebecca; Miller, Jarrett; Pan, Anny; Otoupal, Peter; Alper, Hal S

    2014-01-01

    Economic feasibility of biosynthetic fuel and chemical production hinges upon harnessing metabolism to achieve high titre and yield. Here we report a thorough genotypic and phenotypic optimization of an oleaginous organism to create a strain with significant lipogenesis capability. Specifically, we rewire Yarrowia lipolytica's native metabolism for superior de novo lipogenesis by coupling combinatorial multiplexing of lipogenesis targets with phenotypic induction. We further complete direct conversion of lipid content into biodiesel. Tri-level metabolic control results in saturated cells containing upwards of 90% lipid content and titres exceeding 25 g l(-1) lipids, which represents a 60-fold improvement over parental strain and conditions. Through this rewiring effort, we advance fundamental understanding of lipogenesis, demonstrate non-canonical environmental and intracellular stimuli and uncouple lipogenesis from nitrogen starvation. The high titres and carbon-source independent nature of this lipogenesis in Y. lipolytica highlight the potential of this organism as a platform for efficient oleochemical production. PMID:24445655

  12. Production of omega-3 eicosapentaenoic acid by metabolic engineering of Yarrowia lipolytica.

    PubMed

    Xue, Zhixiong; Sharpe, Pamela L; Hong, Seung-Pyo; Yadav, Narendra S; Xie, Dongming; Short, David R; Damude, Howard G; Rupert, Ross A; Seip, John E; Wang, Jamie; Pollak, Dana W; Bostick, Michael W; Bosak, Melissa D; Macool, Daniel J; Hollerbach, Dieter H; Zhang, Hongxiang; Arcilla, Dennis M; Bledsoe, Sidney A; Croker, Kevin; McCord, Elizabeth F; Tyreus, Bjorn D; Jackson, Ethel N; Zhu, Quinn

    2013-08-01

    The availability of the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) is currently limited because they are produced mainly by marine fisheries that cannot keep pace with the demands of the growing market for these products. A sustainable non-animal source of EPA and DHA is needed. Metabolic engineering of the oleaginous yeast Yarrowia lipolytica resulted in a strain that produced EPA at 15% of dry cell weight. The engineered yeast lipid comprises EPA at 56.6% and saturated fatty acids at less than 5% by weight, which are the highest and the lowest percentages, respectively, among known EPA sources. Inactivation of the peroxisome biogenesis gene PEX10 was crucial in obtaining high EPA yields and may increase the yields of other commercially desirable lipid-related products. This technology platform enables the production of lipids with tailored fatty acid compositions and provides a sustainable source of EPA. PMID:23873085

  13. Bioconversion of volatile fatty acids into lipids by the oleaginous yeast Yarrowia lipolytica.

    PubMed

    Fontanille, Pierre; Kumar, Vinod; Christophe, Gwendoline; Nouaille, Régis; Larroche, Christian

    2012-06-01

    The valorization of volatile fatty acids into microbial lipids by the oleaginous yeast Yarrowia lipolytica was investigated. Therefore, a two-stage fed-batch strategy was designed: the yeast was initially grown on glucose or glycerol as carbon source, then sequential additions of acetic acid under nitrogen limiting conditions were performed after glucose or glycerol exhaustion. The typical values obtained with an initial 40 g/L concentration of glucose were close to 31 g/L biomass, a lipid concentration of 12.4 g/L, which correspond to a lipid content of the biomass close to 40%. This cultivation strategy was also efficient with other volatile fatty acids (butyric and propionic acids) or with a mixture of these three VFAs. The lipids composition was found quite similar to that of vegetable oils. The study demonstrated the feasibility of simultaneous biovalorization of volatile fatty acids and glycerol, two cheap industrial by-products. PMID:22464419

  14. Production of Lycopene in the Non-Carotenoid-Producing Yeast Yarrowia lipolytica

    PubMed Central

    Ketelhot, Markus; Gatter, Michael; Barth, Gerold

    2014-01-01

    The codon-optimized genes crtB and crtI of Pantoea ananatis were expressed in Yarrowia lipolytica under the control of the TEF1 promoter of Y. lipolytica. Additionally, the rate-limiting genes for isoprenoid biosynthesis in Y. lipolytica, GGS1 and HMG1, were overexpressed to increase the production of lycopene. All of the genes were also expressed in a Y. lipolytica strain with POX1 to POX6 and GUT2 deleted, which led to an increase in the size of lipid bodies and a further increase in lycopene production. Lycopene is located mainly within lipid bodies, and increased lipid body formation leads to an increase in the lycopene storage capacity of Y. lipolytica. Growth-limiting conditions increase the specific lycopene content. Finally, a yield of 16 mg g−1 (dry cell weight) was reached in fed-batch cultures, which is the highest value reported so far for a eukaryotic host. PMID:24375130

  15. One-step integration of multiple genes into the oleaginous yeast Yarrowia lipolytica.

    PubMed

    Gao, Shuliang; Han, Linna; Zhu, Li; Ge, Mei; Yang, Sheng; Jiang, Yu; Chen, Daijie

    2014-12-01

    Yarrowia lipolytica is an unconventional yeast, and is generally recognized as safe (GRAS). It provides a versatile fermentation platform that is used commercially to produce many added-value products. Here we report a multiple fragment assembly method that allows one-step integration of an entire β-carotene biosynthesis pathway (~11 kb, consisting of four genes) via in vivo homologous recombination into the rDNA locus of the Y. lipolytica chromosome. The highest efficiency was 21%, and the highest production of β-carotene was 2.2 ± 0.3 mg per g dry cell weight. The total procedure was completed in less than one week, as compared to a previously reported sequential gene integration method that required n weeks for n genes. This time-saving method will facilitate synthetic biology, metabolic engineering and functional genomics studies of Y. lipolytica. PMID:25216641

  16. Immobilizing Yarrowia lipolytica Lipase Lip2 via Improvement of Microspheres by Gelatin Modification.

    PubMed

    Xie, Rong; Cui, Caixia; Chen, Biqiang; Tan, Tianwei

    2015-10-01

    The purpose of this study was to investigate the feasibility of immobilizing Yarrowia lipolytica lipase lip2 on epoxy microspheres with or without gelatin modifications. The activity of lipase immobilized on gelatin-modified supports was twofold higher than those immobilized on native supports. There was no significant difference in the Michaelis-Menten constant (K M ) between the two immobilized lipases. However, lipase immobilized on gelatin modified supports showed an approximately fourfold higher V max than lipase immobilized on native supports. Lipase immobilization on the gelatin-modified support exhibited a significantly improved operational stability in an esterification system. After it was reused for a total of 35 batches, the ester conversion of lipase immobilized on gelatin-modified and native microspheres was 83 and 60 %, respectively. Furthermore, the immobilized lipase could be stored at 4 °C for 12 months without any loss of activity. PMID:26245260

  17. Selection of Yarrowia lipolytica strains with high protein content from yeasts isolated from different marine environments

    NASA Astrophysics Data System (ADS)

    Chi, Zhenming; Wang, Fang; Wang, Lin; Li, Jing; Wang, Xianghong

    2007-10-01

    A total of 78 Yarrowia lipolytica yeast strains from seawater, sediments, mud of salterns, the guts of marine fish, and marine algae were obtained. After the crude protein of the yeasts was estimated by the method of Kjehldahl, we found that seven strains of the marine yeasts grown in soy bean cake hydrolysate with 20 g L-1 of glucose for 48 h at 28°C contained more than 41.0 g protein per 100 g of cell dry weight and the cell dry weight was more than 4.4 g per L of the culture. Among them, strain SWJ-1b contained the highest crude protein. The results of Biolog identification and molecular methods further confirmed that they indeed belonged to Y. lipolytica.

  18. Production of technical-grade sodium citrate from glycerol-containing biodiesel waste by Yarrowia lipolytica.

    PubMed

    Kamzolova, Svetlana V; Vinokurova, Natalia G; Lunina, Julia N; Zelenkova, Nina F; Morgunov, Igor G

    2015-10-01

    The production of technical-grade sodium citrate from the glycerol-containing biodiesel waste by Yarrowia lipolytica was studied. Batch experiments showed that citrate was actively produced within 144 h, then citrate formation decreased presumably due to inhibition of enzymes involved in this process. In contrast, when the method of repeated batch cultivation was used, the formation of citrate continued for more than 500 h. In this case, the final concentration of citrate in the culture liquid reached 79-82 g/L. Trisodium citrate was isolated from the culture liquid filtrate by the addition of a small amount of NaOH, so that the pH of the filtrate increased to 7-8. This simple and economic isolation procedure gave the yield of crude preparation containing trisodium citrate 5.5-hydrate up to 82-86%. PMID:26141285

  19. Mineral supplementation increases erythrose reductase activity in erythritol biosynthesis from glycerol by Yarrowia lipolytica.

    PubMed

    Tomaszewska, Ludwika; Rymowicz, Waldemar; Rywińska, Anita

    2014-03-01

    The aim of this study was to examine the impact of divalent copper, iron, manganese, and zinc ions on the production of erythritol from glycerol by Yarrowia lipolytica and their effect on the activity of erythrose reductase. No inhibitory effect of the examined minerals on yeast growth was observed in the study. Supplementation with MnSO4 · 7H2O (25 mg l(-1)) increased erythritol production by Y. lipolytica by 14.5%. In the bioreactor culture with manganese ion addition, 47.1 g l(-1) of erythritol was produced from 100.0 g l(-1) of glycerol, which corresponded to volumetric productivity of 0.87 g l(-1) h(-1). The addition of Mn(2+) enhanced the intracellular activity of erythrose reductase up to 24.9 U g(-1) of dry weight of biomass (DW), hence, about 1.3 times more than in the control. PMID:24488778

  20. Engineering Yarrowia lipolytica for production of medium-chain fatty acids.

    PubMed

    Rutter, Charles D; Zhang, Shuyan; Rao, Christopher V

    2015-09-01

    Lipids are naturally derived products that offer an attractive, renewable alternative to petroleum-based hydrocarbons. While naturally produced long-chain fatty acids can replace some petroleum analogs, medium-chain fatty acid would more closely match the desired physical and chemical properties of currently employed petroleum products. In this study, we engineered Yarrowia lipolytica, an oleaginous yeast that naturally produces lipids at high titers, to produce medium-chain fatty acids. Five different acyl-acyl carrier protein (ACP) thioesterases with specificity for medium-chain acyl-ACP molecules were expressed in Y. lipolytica, resulting in formation of either decanoic or octanoic acid. These novel fatty acid products were found to comprise up to 40 % of the total cell lipids. Furthermore, the reduction in chain length resulted in a twofold increase in specific lipid productivity in these engineered strains. The medium-chain fatty acids were found to be incorporated into all lipid classes. PMID:26129951

  1. Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants.

    PubMed

    Rakicka, Magdalena; Rywińska, Anita; Cybulski, Krzysztof; Rymowicz, Waldemar

    2016-01-01

    Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300gL(-1)), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142gL(-1) after 5 days, which corresponded to 0.47gg(-1) yield and productivity of 1.1gL(-1)h(-1). Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol. PMID:26991292

  2. Grundlagen der Mechatronik

    NASA Astrophysics Data System (ADS)

    Roddeck, Werner

    Der Begriff Mechatronik ist ein Kunstwort, welches durch Eindeutschung des englischen Wortes "Mechatronics“ entstanden ist. Dieses ist wiederum eine Zusammenziehung der englischen Bezeichnungen für "Mechanics“ (Maschinenbau) und "Electronics“ (Elektrotechnik). Der Begriff wurde durch einen japanischen Ingenieur 1969 geprägt und durch eine japanische Firma bis 1972 als Warenzeichen gehalten.

  3. Der II. Hauptsatz der Wärmelehre

    NASA Astrophysics Data System (ADS)

    Heintze, Joachim

    Wir haben in (4.44) den II. Hauptsatz als empirische Tatsache folgendermaßen formuliert: (i) Wärmeenergie geht von selbst nur von einem wärmeren Körper auf einen kälteren über, niemals in der umgekehrten Richtung. Nun werden wir beweisen, dass sich aus diesem Prinzip folgende äquivalente Formulierungen für den II. Hauptsatz ableiten lassen: (ii) Es ist unmöglich, ein Perpetuum mobile zweiter Art zu bauen, d. h. eine Maschine, die fortlaufend Wärmeenergie vollständig in mechanische Arbeit umsetzen kann. Eine Wärmekraftmaschine, die einen Kreisprozess mit der höchsten Temperatur Tw und der niedrigsten Temperatur Tk durchläuft, hat höchstens den Carnotschen Wirkungsgrad c = (Tw - Tk)/Tw. Wenn in der Maschine nur reversible Prozesse ablaufen, die gesamte Wärmezufuhr bei der Temperatur Tw erfolgt und ausschließlich bei der Temperatur Tw gekühlt wird, ist ihr Wirkungsgrad = C. Es gibt keine Wärmekraftmaschine, die eine bessere Ausnutzung der Wärmeenergie ermöglicht. (iv) In jedem thermodynamischen System existiert die Zustandsgröße Entropie, definiert durch ihr Differential dS = (dQrev)/T . Entropie kann erzeugt, aber nicht vernichtet werden. Bei Zustandsänderungen, die in einem abgeschlossenen System ablaufen, nimmt die Entropie entweder zu (irreversible Prozesse), oder sie bleibt konstant (reversible Prozesse). Im Anschluss an (iii) werden wir zur Definition der thermodynamischen Temperatur und bei der Diskussion von (iv) zu einem tieferen Verständnis der Entropie gelangen. Es zeigt sich, dass die Entropie das eigentliche Bindeglied zwischen Mechanik und Wärmelehre darstellt. Am Ende des Kapitels werden wir einige Anwendungen des II. Hauptsatzes betrachten.

  4. Kants Theorie der Sonne: Physikgeschichte

    NASA Astrophysics Data System (ADS)

    Jacobi, Manfred

    2005-01-01

    Im Rahmen seiner Kosmogonie entwickelte der junge Immanuel Kant eine Theorie der Sonne. Sie ist ein einzigartiges Zeugnis seiner intuitiven Vorstellungskraft und beweist auch die Leistungsfähigkeit der damaligen, vorwiegend von Newton geprägten Weltsicht. Entstehung, Aufbau und Dynamik der Sonne werden in Kants Theorie ebenso erklärt wie etwa das Phänomen der Sonnenflecken.

  5. High-throughput transformation method for Yarrowia lipolytica mutant library screening.

    PubMed

    Leplat, Christophe; Nicaud, Jean-Marc; Rossignol, Tristan

    2015-09-01

    As a microorganism of major biotechnological importance, the oleaginous yeast Yarrowia lipolytica is subjected to intensive genetic engineering and functional genomic analysis. Future advancements in this area, however, require a system that will generate a large collection of mutants for high-throughput screening. Here, we report a rapid and efficient method for high-throughput transformation of Y. lipolytica in 96-well plates. We developed plasmids and strains for the large-scale screening of overexpression mutant strains, using Gateway® vectors that were adapted for specific locus integration in Y. lipolytica. As an example, a collection of mutants that overexpressed the alkaline extracellular protease (AEP) was obtained in a single transformation experiment. The platform strain that we developed to receive the overexpression cassette was designed to constitutively express a fluorescent protein as a convenient growth reporter for screening in non-translucid media. An example of growth comparison in skim milk-based medium between AEP overexpression and deletion mutants is provided. PMID:26100263

  6. An evolutionary metabolic engineering approach for enhancing lipogenesis in Yarrowia lipolytica.

    PubMed

    Liu, Leqian; Pan, Anny; Spofford, Caitlin; Zhou, Nijia; Alper, Hal S

    2015-05-01

    Lipogenic organisms provide an ideal platform for biodiesel and oleochemical production. Through our previous rational metabolic engineering efforts, lipogenesis titers in Yarrowia lipolytica were significantly enhanced. However, the resulting strain still suffered from decreased biomass generation rates. Here, we employ a rapid evolutionary metabolic engineering approach linked with a floating cell enrichment process to improve lipogenesis rates, titers, and yields. Through this iterative process, we were able to ultimately improve yields from our prior strain by 55% to achieve production titers of 39.1g/L with upwards of 76% of the theoretical maximum yield of conversation. Isolated cells were saturated with up to 87% lipid content. An average specific productivity of 0.56g/L/h was achieved with a maximum instantaneous specific productivity of 0.89g/L/h during the lipid production phase in fermentation. Genomic sequencing of the evolved strains revealed a link between a decrease/loss of function mutation of succinate semialdehyde dehydrogenase, uga2, suggesting the importance of gamma-aminobutyric acid assimilation in lipogenesis. This linkage was validated through gene deletion experiments. This work presents an improved host strain that can serve as a platform for efficient oleochemical production. PMID:25724340

  7. A new recombinant protein expression system for high-throughput screening in the yeast Yarrowia lipolytica.

    PubMed

    Bordes, Florence; Fudalej, Franck; Dossat, Valérie; Nicaud, Jean-Marc; Marty, Alain

    2007-09-01

    Development of a high-throughput eukaryotic screening procedure is important to increase success in obtaining improved enzymes through directed enzyme evolution. This procedure was developed for the yeast Yarrowia lipolytica which becomes the second eukaryotic host for this purpose. The extracellular lipase Lip2 was used as expressed enzyme but this system will be easily adjusted for other enzymes. We adapted and optimized the protocol for protein expression by Y. lipolytica in 96-well microplates. Yeast transformation efficiency and expression cassette insertion were increased by constructing a strain containing a zeta docking platform for targeted integration into the genome. The coefficient of variance of the full process was reduced from 36.3% to 18.9%. The main part of the variability (11.7%) arises from the specific lipase enzyme assay whereas the coefficient of variance concerning transformation, growth and expression steps represents only 7.2%. The rate of clone with no activity was reduced from 5.8% to 0.2%. Both transformation efficiency and variability are then compatible with high-throughput screening in the yeast Y. lipolytica. PMID:17669530

  8. Engineering and production of laccase from Trametes versicolor in the yeast Yarrowia lipolytica.

    PubMed

    Theerachat, Monnat; Emond, Stéphane; Cambon, Emmanuelle; Bordes, Florence; Marty, Alain; Nicaud, Jean-Marc; Chulalaksananukul, Warawut; Guieysse, David; Remaud-Siméon, Magali; Morel, Sandrine

    2012-12-01

    The lcc1 gene coding for the laccase from Trametes versicolor DSM11269 was cloned into the genome of Yarrowia lipolytica using either single or multiple integration sites. The levels of the recombinant laccase activity secreted in the culture media were 0.25 and 1 U ml(-1) for single and multiple integrations, respectively. The strain with a single integration was successfully used to express variant libraries which were screened on ABTS substrate. The strain encoding the double mutant L185P/Q214K (rM4A) showed a sixfold enhancement in secreted enzyme activity. The catalytic efficiency of the purified rM-4A laccase was respectively increased 2.4- and 2.8-fold towards ABTS and 2,6-dimethoxyphenol, compared to the rWT. Culture supernatants containing either rWT or rM-4A catalyzed the almost complete decolorization of an Amaranth solution (70 nMs(-1)). Taken together, our results open new perspectives for the use of Y. lipolytica as a molecular evolution platform to engineer laccases with improved properties. PMID:23026343

  9. [The Engineering of a Yarrowia lipolytica Yeast Strain Capable of Homologous Recombination of the Mitochondrial Genome].

    PubMed

    Isakova, E P; Epova, E Yu; Sekova, V Yu; Trubnikova, E V; Kudykina, Yu K; Zylkova, M V; Guseva, M A; Deryabina, Yu I

    2015-01-01

    None of the studied eukaryotic species has a natural system for homologous recombination of the mitochondrial genome. We propose an integrated genetic construct pQ-SRUS, which allows introduction of the recA gene from Bacillus subtilis into the nuclear genome of an extremophilic yeast, Yarrowia lipolytica. The targeting of recombinant RecA to the yeast mitochondria is provided by leader sequences (5'-UTR and 3'-UTR) derived from the SOD2 gene mRNA, which exhibits affinity to the outer mitochondrial membrane and thus provides cotranslational transport of RecA to the inner space of the mitochondria. The Y. lipolytica strain bearing the pQ-SRUS construct has the unique ability to integrate DNA constructs into the mitochondrial genome. This fact was confirmed using a tester construct, pQ-NIHN, intended for the introduction of the EYFP gene into the translation initiation region of the Y. lipolytica ND1 mitochondrial gene. The Y. lipolytica strain bearing pQ-SRUS makes it possible to engineer recombinant producers based on Y. lipolytica bearing transgenes in the mitochondrial genome. They are promising for the construction of a genetic system for in vivo replication and modification of the human mitochondrial genome. These strains may be used as a tool for the treatment of human mitochondrial diseases (including genetically inherited ones). PMID:26204776

  10. [Biosynthesis of isocitric acid by the yeast yarrowia lipolytica and its regulation].

    PubMed

    Kamzolova, S V; Lunin, Y N; Allayarov, R K; Puntus, I F; Laptev, I A; Samojlenko, V A; Morgunov, I G

    2015-01-01

    We studied the biosynthesis of isocitric acid from rapeseed (canola) oil by the yeast Yarrowia lipolytica and its regulation. We determined a fundamental possibility for directed biosynthesis of isocitric acid by Y lipolytica yeast, with only minimal amounts of citric acid byproduct, when grown on a medium containing canola oil. Wild type strains of Y lipolytica were mutagenized by UV irradiation and treatment with N-methyl-N'-nitro-N-nitrosoguanidine (NG). Subsequent selection on media with acetate and isocitrate resulted in isolation of a UV/NG Y lipolytica UV/NG mutant that synthesized isocitrate and citrate at a ratio of 2.7:1. In the parent strain, this ratio is 1:1. Inhibition of isocitrate lyase, a key enzyme in the metabolism of isocitric acid, by the addition of itaconic acid resulted in increased synthesis of isocitrate with a ratio of isocitrate to citrate reaching 6:1. Culturing of the Y lipolytica UV/NG mutant in a pilot industrial fermenter in the presence of itaconic acid resulted in the production of 88.7 g/L of isocitric acid with a yield of 90%. PMID:26027362

  11. Bioconversion of R-(+)-limonene to perillic acid by the yeast Yarrowia lipolytica

    PubMed Central

    Ferrara, Maria Antonieta; Almeida, Débora S.; Siani, Antonio C.; Lucchetti, Leonardo; Lacerda, Paulo S.B.; Freitas, André; Tappin, Marcelo R.R.; Bon, Elba P.S.

    2013-01-01

    Perillyl derivatives are increasingly important due to their flavouring and antimicrobial properties as well as their potential as anticancer agents. These terpenoid species, which are present in limited amounts in plants, may be obtained via bioconversion of selected monoterpene hydrocarbons. In this study, seventeen yeast strains were screened for their ability to oxidize the exocyclic methyl group in the p-menthene moiety of limonene into perillic acid. Of the yeast tested, the highest efficiency was observed for Yarrowia lipolytica ATCC 18942. The conversion of R (+)-limonene by Y. lipolytica was evaluated by varying the pH (3 to 8) and the temperature (25 to 30 °C) in a reaction medium containing 0.5% v/v limonene and 10 g/L of stationary phase cells (dry weight). The best results, corresponding to 564 mg/L of perillic acid, were obtained in buffered medium at pH 7.1 that was incubated at 25 °C for 48 h. The stepwise addition of limonene increased the perillic acid concentration by over 50%, reaching 855 mg/L, whereas the addition of glucose or surfactant to the reaction medium did not improve the bioconversion process. The use of Y. lipolytica showed promise for ease of further downstream processing, as perillic acid was the sole oxidised product of the bioconversion reaction. Moreover, bioprocesses using safe and easy to cultivate yeast cells have been favoured in industry. PMID:24688495

  12. Yarrowia lipolytica as a biotechnological chassis to produce usual and unusual fatty acids.

    PubMed

    Ledesma-Amaro, Rodrigo; Nicaud, Jean-Marc

    2016-01-01

    One of the most promising alternatives to petroleum for the production of fuels and chemicals is bio-oil based chemistry. Microbial oils are gaining importance because they can be engineered to accumulate lipids enriched in desired fatty acids. These specific lipids are closer to the commercialized product, therefore reducing pollutants and costly chemical steps. Yarrowia lipolytica is the most widely studied and engineered oleaginous yeast. Different molecular and bioinformatics tools permit systems metabolic engineering strategies in this yeast, which can produce usual and unusual fatty acids. Usual fatty acids, those usually found in triacylglycerol, accumulate through the action of several pathways, such as fatty acid/triacylglycerol synthesis, transport and degradation. Unusual fatty acids are enzymatic modifications of usual fatty acids to produce compounds that are not naturally synthetized in the host. Recently, the metabolic engineering of microorganisms has produced different unusual fatty acids, such as building block ricinoleic acid and nutraceuticals such as conjugated linoleic acid or polyunsaturated fatty acids. Additionally, microbial sources are preferred hosts for the production of fatty acid-derived compounds such as γ-decalactone, hexanal and dicarboxylic acids. The variety of lipids produced by oleaginous microorganisms is expected to rise in the coming years to cope with the increasing demand. PMID:26703186

  13. High-level production of extracellular lipase by Yarrowia lipolytica mutants from methyl oleate.

    PubMed

    Darvishi, Farshad; Destain, Jacqueline; Nahvi, Iraj; Thonart, Philippe; Zarkesh-Esfahani, Hamid

    2011-10-01

    The yeast Yarrowia lipolytica degrades efficiently low-cost hydrophobic substrates for the production of various added-value products such as lipases. To obtain yeast strains producing high levels of extracellular lipase, Y. lipolytica DSM3286 was subjected to mutation using ethyl methanesulfonate (EMS) and ultraviolet (UV) light. Twenty mutants were selected out of 1600 mutants of Y. lipolytica treated with EMS and UV based on lipase production ability on selective medium. A new industrial medium containing methyl oleate was optimized for lipase production. In the 20 L bioreactor containing new industrial medium, one UV mutant (U6) produced 356 U/mL of lipase after 24h, which is about 10.5-fold higher than that produced by the wild type strain. The properties of the mutant lipase were the same as those of the wild type: molecular weight 38 kDa, optimum temperature 37°C and optimum pH 7. Furthermore, the nucleotide sequences of extracellular lipase gene (LIP2) in wild type and mutant strains were determined. Only two silent substitutions at 362 and 385 positions were observed in the ORF region of LIP2. Two single substitutions and two duplications of the T nucleotide were also detected in the promoter region. LIP2 sequence comparison of the Y. lipolytica DSM3286 and U6 strains shows good targets to effective DNA recombinant for extracellular lipase of Y. lipolytica. PMID:21324386

  14. The production of succinic acid by yeast Yarrowia lipolytica through a two-step process.

    PubMed

    Kamzolova, Svetlana V; Vinokurova, Natalia G; Shemshura, Olga N; Bekmakhanova, Nadiya E; Lunina, Julia N; Samoilenko, Vladimir A; Morgunov, Igor G

    2014-09-01

    The production of α-ketoglutaric acid by yeast Yarrowia lipolytica VKMY-2412 from ethanol and its subsequent chemical conversion to succinic acid (SA) were investigated. A highly effective and environmentally friendly process of α-ketoglutaric acid production was developed using a special pH-controlling strategy, in which the titration of the culture broth with KOH in the acid-formation phase was minimal, that allowed accumulation of only low amounts of inorganic wastes in the course of SA recovery. The culture broth filtrate containing α-ketoglutaric acid (88.7 g l(-1)) was directly employed for SA production; the amount of SA produced comprised 71.7 g l(-1) with the yield of 70% from ethanol consumed. SA was isolated from the culture broth filtrate in a crystalline form with the purity of 100%. The yield of isolated SA was as high as 72% of its amount in the culture broth filtrate. The antimicrobial and nematocidic effects of SA of microbial origin on pathogenic organisms that cause human and plant diseases were revealed for the first time. PMID:24972816

  15. Oil Production from Yarrowia lipolytica Po1g Using Rice Bran Hydrolysate

    PubMed Central

    Tsigie, Yeshitila Asteraye; Wang, Chun-Yuan; Kasim, Novy S.; Diem, Quy-Do; Huynh, Lien-Huong; Ho, Quoc-Phong; Truong, Chi-Thanh; Ju, Yi-Hsu

    2012-01-01

    The purpose of this study was to produce microbial oil from Yarrowia lipolytica Po1g grown in defatted rice bran hydrolysate. After removing oil from rice bran by Soxhlet extraction, the bran is subjected to acid hydrolysis with various sulfuric acid concentrations (1–4% v/v), reaction times (1–8 h), and reaction temperatures (60–120°C). The optimal conditions for maximum total sugar production from the hydrolysate were found to be 3% sulfuric acid at 90°C for 6 h. Glucose was the predominant sugar (43.20 ± 0.28 g/L) followed by xylose (4.93 ± 0.03 g/L) and arabinose (2.09 ± 0.01 g/L). The hydrolysate was subsequently detoxified by neutralization to reduce the amount of inhibitors such as 5-hydroxymethylfurfural (HMF) and furfural to increase its potential as a medium for culturing Y. lipolytica Po1g. Dry cell mass and lipid content of Y. lipolytica Po1g grown in detoxified defatted rice bran hydrolysate (DRBH) under optimum conditions were 10.75 g/L and 48.02%, respectively. PMID:22496604

  16. Animal Bones Char Solubilization by Gel-Entrapped Yarrowia lipolytica on Glycerol-Based Media

    PubMed Central

    Vassileva, Maria; Eichler-Lobermann, Bettina; Reyes, Antonia; Vassilev, Nikolay

    2012-01-01

    Citric acid was produced with free and k-carrageenan-entrapped cells of the yeast Yarrowia lipolytica in single and repeated batch-shake-flask fermentations on glycerol-based media. Simultaneous solubilization of hydroxyapatite of animal bone origin (HABO) was tested in all experiments. The highest citric acid production by free yeast cells of 20.4 g/L and 18.7 g/L was reached after 96 h of fermentation in the absence and presence of 3 g/L HABO, respectively. The maximum values for the same parameter achieved by gel-entrapped cells in conditions of single batch and repeated-batch fermentation processes were 18.7 g/L and 28.1 g/L registered after 96 h and the 3d batch cycle, respectively. The highest citric acid productivity of 0.58 g L−1 h−1 was obtained with immobilized cells in repeated batch mode of fermentation when the added hydroxyapatite of 3 g/L was solubilized to 399 mg/L whereas the maximum efficiency of 89.0% was obtained with 1 g/L of HABO. PMID:22645462

  17. Metabolome analysis and pathway abundance profiling of Yarrowia lipolytica cultivated on different carbon sources.

    PubMed

    Zhao, Chen; Gu, Deqing; Nambou, Komi; Wei, Liujing; Chen, Jun; Imanaka, Tadayuki; Hua, Qiang

    2015-07-20

    Yarrowia lipolytica, a model microorganism of oleaginous yeasts with developed sophisticated genetic tools, is able to metabolize a wide range of substrates and accumulate large amounts of lipids. However, there is a lack of literature reporting the metabolic characteristics of Y. lipolytica metabolizing these substrates in a systematic view. In this study, Y. lipolytica was cultivated on a variety of carbon sources, among which cell growth and production characteristics on two representative substrates (glucose and oleic acid) were investigated in detail at metabolomic level. Metabolic pathway abundance was computed to interpret the metabolome data in a straightforward way. The results showed that most pathway abundances decreased in the shift from growth to production phase. Specifically, when cultivated on glucose, abundances of twelve pathways decreased markedly between the growth and lipid production phases, while thirteen pathways reduced and only three pathways increased significantly in abundances on oleic acid. In comparison, for the same cultivation phase only a few pathways exhibited significant changes between glucose-grown and oleic acid-grown cells. This study revealed that the pathway abundance could be used to effectively show the activity changes of pathways, providing a new perspective to employ metabolomics data for understanding cell metabolism and enhancing the production of target metabolites. PMID:25912211

  18. Highly Efficient Fructooligosaccharides Production by an Erythritol-Producing Yeast Yarrowia lipolytica Displaying Fructosyltransferase.

    PubMed

    Zhang, Lebin; An, Jin; Li, Lijuan; Wang, Hengwei; Liu, Dawen; Li, Ning; Cheng, Hairong; Deng, Zixin

    2016-05-18

    Currently, fructooligosaccharides (FOS) are industrially transformed from sucrose by purified enzymes or fungi cells. However, these methods are expensive and time-consuming. An economical approach to producing FOS using erythritol-producing yeast cells was described in this study. Fructosyltransferase from Aspergillus oryzae was displayed on the cell surface of Yarrowia lipolytica, resulting in an engineered strain capable of transforming sucrose to FOS. An amount of 480 g/L FOS was produced within 3 h in a solution of 800 g/L sucrose and 5 g/L cells (dry cell weight, DCW) at pH 6.0 and 60 °C, with a yield of 60% of total sucrose and a productivity of 160 g/(L·h). The yeast pastes from the erythritol industry can be repeatedly used as the whole-cell catalysts at least 10 times by this newly developed approach. This efficient method is attractive for the large-scale production of FOS from sucrose. PMID:27124471

  19. Quantitative image analysis as a tool for Yarrowia lipolytica dimorphic growth evaluation in different culture media.

    PubMed

    Braga, A; Mesquita, D P; Amaral, A L; Ferreira, E C; Belo, I

    2016-01-10

    Yarrowia lipolytica, a yeast strain with a huge biotechnological potential, capable to produce metabolites such as γ-decalactone, citric acid, intracellular lipids and enzymes, possesses the ability to change its morphology in response to environmental conditions. In the present study, a quantitative image analysis (QIA) procedure was developed for the identification and quantification of Y. lipolytica W29 and MTLY40-2P strains dimorphic growth, cultivated in batch cultures on hydrophilic (glucose and N-acetylglucosamine (GlcNAc) and hydrophobic (olive oil and castor oil) media. The morphological characterization of yeast cells by QIA techniques revealed that hydrophobic carbon sources, namely castor oil, should be preferred for both strains growth in the yeast single cell morphotype. On the other hand, hydrophilic sugars, namely glucose and GlcNAc caused a dimorphic transition growth towards the hyphae morphotype. Experiments for γ-decalactone production with MTLY40-2P strain in two distinct morphotypes (yeast single cells and hyphae cells) were also performed. The obtained results showed the adequacy of the proposed morphology monitoring tool in relation to each morphotype on the aroma production ability. The present work allowed establishing that QIA techniques can be a valuable tool for the identification of the best culture conditions for industrial processes implementation. PMID:26546055

  20. Tuning Gene Expression in Yarrowia lipolytica by a Hybrid Promoter Approach▿†

    PubMed Central

    Blazeck, John; Liu, Leqian; Redden, Heidi; Alper, Hal

    2011-01-01

    The development of strong and tunable promoter elements is necessary to enable metabolic and pathway engineering applications for any host organism. Here, we have expanded and generalized a hybrid promoter approach to produce libraries of high-expressing, tunable promoters in the nonconventional yeast Yarrowia lipolytica. These synthetic promoters are comprised of two modular components: the enhancer element and the core promoter element. By exploiting this basic promoter architecture, we have overcome native expression limitations and provided a strategy for both increasing the native promoter capacity and producing libraries for tunable gene expression in a cellular system with ill-defined genetic tools. In doing so, this work has created the strongest promoters ever reported for Y. lipolytica. Furthermore, we have characterized these promoters at the single-cell level through the use of a developed fluorescence-based assay as well as at the transcriptional and whole-cell levels. The resulting promoter libraries exhibited a range of more than 400-fold in terms of mRNA levels, and the strongest promoters in this set had 8-fold-higher fluorescence levels than those of typically used endogenous promoters. These results suggest that promoters in Y. lipolytica are enhancer limited and that this limitation can be partially or fully alleviated through the addition of tandem copies of upstream activation sequences (UASs). Finally, this work illustrates that tandem copies of UAS regions can serve as synthetic transcriptional amplifiers that may be generically used to increase the expression levels of promoters. PMID:21926196

  1. Morphogenetic behavior of tropical marine yeast Yarrowia lipolytica in response to hydrophobic substrates.

    PubMed

    Zinjarde, Smita S; Kale, Bhagyashree V; Vishwasrao, Paresh V; Kumar, Ameeta R

    2008-09-01

    The morphogenetic behavior of a tropical marine Yarrowia lipolytica strain on hydrophobic substrates was studied. Media containing coconut oil or palm kernel oil (rich in lauric and myristic acids) prepared in distilled water or seawater at a neutral pH supported 95% of the cells to undergo a transition from the yeast form to the mycelium form. With potassium laurate, 51% of the cells were in the mycelium form, whereas with myristate, 32% were in the mycelium form. However, combinations of these two fatty acids in proportions that are present in coconut oil or palm kernel oil enhanced the mycelium formation to 65%. The culture also produced extracellular lipases during the morphogenetic change. The yeast cells were found to attach to the large droplets of the hydrophobic substrates during the transition, while the mycelia were associated with the aqueous phase. The alkane-grown yeast partitioned more efficiently in the hydrophobic phases when compared with the coconut oil-grown mycelia. A fatty acid analysis of the mycelial form revealed the presence of lauric acid in addition to the long-chain saturated and unsaturated fatty acids observed in the yeast form. The mycelia underwent a rapid transition to the yeast form with n-dodecane, a medium-chain aliphatic hydrocarbon. Thus, the fungus displayed a differential behavior towards the two types of saturated hydrophobic substrates. PMID:18852507

  2. Biotechnological production of γ-decalactone, a peach like aroma, by Yarrowia lipolytica.

    PubMed

    Braga, A; Belo, I

    2016-10-01

    The request for new flavourings increases every year. Consumer perception that everything natural is better is causing an increase demand for natural aroma additives. Biotechnology has become a way to get natural products. γ-Decalactone is a peach-like aroma widely used in dairy products, beverages and others food industries. In more recent years, more and more studies and industrial processes were endorsed to cost-effect this compound production. One of the best-known methods to produce γ-decalactone is from ricinoleic acid catalyzed by Yarrowia lipolytica, a generally regarded as safe status yeast. As yet, several factors affecting γ-decalactone production remain to be fully understood and optimized. In this review, we focus on the aromatic compound γ-decalactone and its production by Y. lipolytica. The metabolic pathway of lactone production and degradation are addressed. Critical analysis of novel strategies of bioprocess engineering, metabolic and genetic engineering and other strategies for the enhancement of the aroma productivity are presented. PMID:27565779

  3. The peculiarities of succinic acid production from rapeseed oil by Yarrowia lipolytica yeast.

    PubMed

    Kamzolova, Svetlana V; Vinokurova, Natalia G; Dedyukhina, Emiliya G; Samoilenko, Vladimir A; Lunina, Julia N; Mironov, Alexey A; Allayarov, Ramil K; Morgunov, Igor G

    2014-05-01

    The process of succinic acid (SA) production represents the combination of microbial synthesis of α-ketoglutaric acid from rapeseed oil by yeast Yarrowia lipolytica VKM Y-2412 and subsequent decarboxylation of α-ketoglutaric acid by hydrogen peroxide to SA that leads to the production of 69.0 g l(-1) of SA and 1.36 g l(-1) of acetic acid. SA was isolated from the culture broth filtrate in a crystalline form. The SA recovery from the culture filtrate has certain difficulties due to the presence of residual triglycerides of rapeseed oil. The effect of different methods of the culture filtrate treatment and various sorption materials on the coagulation of triglycerides was studied, and as a result, the precipitation of residual triglycerides by acetone was chosen. The subsequent isolation procedures involved the decomposition of H2O2 in the filtrate followed by filtrate bleaching and acidification with a mineral acid, evaporation of filtrate, and SA extraction with ethanol from the residue. The purity of crystalline SA isolated from the culture broth filtrate achieved 97.6-100 %. The product yield varied from 62.6 to 71.6 % depending on the acidity of the supernatant. PMID:24531240

  4. A comparative study on glycerol metabolism to erythritol and citric acid in Yarrowia lipolytica yeast cells.

    PubMed

    Tomaszewska, Ludwika; Rakicka, Magdalena; Rymowicz, Waldemar; Rywińska, Anita

    2014-09-01

    Citric acid and erythritol biosynthesis from pure and crude glycerol by three acetate-negative mutants of Yarrowia lipolytica yeast was investigated in batch cultures in a wide pH range (3.0-6.5). Citric acid biosynthesis was the most effective at pH 5.0-5.5 in the case of Wratislavia 1.31 and Wratislavia AWG7. With a decreasing pH value, the direction of biosynthesis changed into erythritol synthesis accompanied by low production of citric acid. Pathways of glycerol conversion into erythritol and citric acid were investigated in Wratislavia K1 cells. Enzymatic activity was compared in cultures run at pH 3.0 and 4.5, that is, under conditions promoting the production of erythritol and citric acid, respectively. The effect of pH value (3.0 and 4.5) and NaCl presence on the extracellular production and intracellular accumulation of citric acid and erythritol was compared as well. Low pH and NaCl resulted in diminished activity of glycerol kinase, whereas such conditions stimulated the activity of glycerol-3-phosphate dehydrogenase. The presence of NaCl strongly influenced enzymes activity - the effective erythritol production was correlated with a high activity of transketolase and erythrose reductase. Therefore, presented results confirmed that transketolase and erythrose reductase are involved in the overproduction of erythritol in the cells of Y. lipolytica yeast. PMID:25041612

  5. Enhanced production of erythritol by Yarrowia lipolytica on glycerol in repeated batch cultures.

    PubMed

    Mirończuk, Aleksandra M; Furgała, Joanna; Rakicka, Magdalena; Rymowicz, Waldemar

    2014-01-01

    Erythritol is an important natural sweetener, industrially produced only by fermentation on glucose media. Glycerol is an important renewable feedstock as it is the major by-product of the biodiesel production process; here we present an alternative way to convert this low-cost substrate into value-added products, such as erythritol. Repeated batch cultures (RBC) were performed to improve the productivity of erythritol from pure and crude glycerol. An acetate negative mutant of Yarrowia lipolytica Wratislavia K1 was found to be applicable for the production of high amounts of erythritol in RBC. When 20 % of fresh replaced medium was added, the strain Wratislavia K1 was able to produce 220 g l (-1) erythritol, which corresponded to a 0.43 g g(-1) yield and a productivity of 0.54 g l(-1) h(-1). Additionally, the activity of the culture remained stable for more than 1,000 h, i.e., 11 cycles of the repeated batch bioreactors. PMID:24281394

  6. Optimization of medium composition for erythritol production from glycerol by Yarrowia lipolytica using response surface methodology.

    PubMed

    Rywińska, Anita; Marcinkiewicz, Marta; Cibis, Edmund; Rymowicz, Waldemar

    2015-08-18

    Several factors affecting erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 strain were examined in batch fermentations. Ammonium sulfate, monopotassium phosphate, and sodium chloride were identified as critical medium components that determine the ratio of polyols produced. The central composite rotatable experimental design was used to optimize medium composition for erythritol production. The concentrations of ammonium sulfate, monopotassium phosphate, and sodium chloride in the optimized medium were 2.25, 0.22, and 26.4 g L(-1), respectively. The C:N ratio was found as 81:1. In the optimized medium with 100 g L(-1) of glycerol the Wratislavia K1 strain produced 46.9 g L(-1) of erythritol, which corresponded to a 0.47 g g(-1) yield and a productivity of 0.85 g L(-1) hr(-1). In the fed-batch mode and medium with the total concentration of glycerol at 300 g L(-1) and C:N ratio at 81:1, 132 g L(-1) of erythritol was produced with 0.44 g g(-1) yield and a productivity of 1.01 g L(-1) hr(-1.) PMID:25387364

  7. [Redox Status of Extremophilic Yeast Yarrowia Lipolytica During Adaptation to pH-Stress].

    PubMed

    Sekova, V Y; Gessler, N N; Isakova, E P; Antipov, A N; Dergacheva, D I; Deryabina, Y I; Trubnikova, E V

    2015-01-01

    In this study we investigated the activities of antioxidant enzymes (superoxide dismutases (SODs) and catalases (CATs)) and the ROS level in cells of Yarrowia lipolytica yeasts grown in a medium with different pH values (4.5, 5.5 and 9.0). It was shown that an increase in the cellular ROS level took place under both acid and alkaline conditions. The growth under extreme conditions was accompanied by a significant increase of SOD activity (by 2.5 times in the acid medium and by 4 times in the alkaline medium), but catalase activity did not change. A study of the electrophoretic profile of catalases showed the presence of three isoforms differing in inhibitor resistance. The electrophoretic profiles of SODs and their inhibitory analysis revealed there are two other isoforms, probably of mitochondrial origin, in addition to Cu and Zn SOD. The role of SOD in pH-adaptation of extremophilic Y. lipolytica yeasts is discussed. PMID:26859958

  8. Citric acid production in Yarrowia lipolytica SWJ-1b yeast when grown on waste cooking oil.

    PubMed

    Liu, Xiaoyan; Lv, Jinshun; Xu, Jiaxing; Zhang, Tong; Deng, Yuanfang; He, Jianlong

    2015-03-01

    In this study, citric acid was produced from waste cooking oil by Yarrowia lipolytica SWJ-1b. To get the maximal yield of citric acid, the compositions of the medium for citric acid production were optimized, and our results showed that extra nitrogen and magnesium rather than vitamin B1 and phosphate were needed for CA accumulation when using waste cooking oil. The results also indicated that the optimal initial concentration of the waste cooking oil in the medium for citric acid production was 80.0 g/l, and the ideal inoculation size was 1 × 10(7) cells/l of medium. We also reported that during 10-l fermentation, 31.7 g/l of citric acid, 6.5 g/l of isocitric acid, 5.9 g/l of biomass, and 42.1 g/100.0 g cell dry weight of lipid were attained from 80.0 g/l of waste cooking oil within 336 h. At the end of the fermentation, 94.6 % of the waste cooking oil was utilized by the cells of Y. lipolytica SWJ-1b, and the yield of citric acid was 0.4 g/g waste cooking oil, which suggested that waste cooking oil was a suitable carbon resource for citric acid production. PMID:25488499

  9. Multiplex gene editing of the Yarrowia lipolytica genome using the CRISPR-Cas9 system.

    PubMed

    Gao, Shuliang; Tong, Yangyang; Wen, Zhiqiang; Zhu, Li; Ge, Mei; Chen, Daijie; Jiang, Yu; Yang, Sheng

    2016-08-01

    Yarrowia lipolytica is categorized as a generally recognized as safe (GRAS) organism and is a heavily documented, unconventional yeast that has been widely incorporated into multiple industrial fields to produce valuable biochemicals. This study describes the construction of a CRISPR-Cas9 system for genome editing in Y. lipolytica using a single plasmid (pCAS1yl or pCAS2yl) to transport Cas9 and relevant guide RNA expression cassettes, with or without donor DNA, to target genes. Two Cas9 target genes, TRP1 and PEX10, were repaired by non-homologous end-joining (NHEJ) or homologous recombination, with maximal efficiencies in Y. lipolytica of 85.6 % for the wild-type strain and 94.1 % for the ku70/ku80 double-deficient strain, within 4 days. Simultaneous double and triple multigene editing was achieved with pCAS1yl by NHEJ, with efficiencies of 36.7 or 19.3 %, respectively, and the pCASyl system was successfully expanded to different Y. lipolytica breeding strains. This timesaving method will enable and improve synthetic biology, metabolic engineering and functional genomic studies of Y. lipolytica. PMID:27349768

  10. Oil production from Yarrowia lipolytica Po1g using rice bran hydrolysate.

    PubMed

    Tsigie, Yeshitila Asteraye; Wang, Chun-Yuan; Kasim, Novy S; Diem, Quy-Do; Huynh, Lien-Huong; Ho, Quoc-Phong; Truong, Chi-Thanh; Ju, Yi-Hsu

    2012-01-01

    The purpose of this study was to produce microbial oil from Yarrowia lipolytica Po1g grown in defatted rice bran hydrolysate. After removing oil from rice bran by Soxhlet extraction, the bran is subjected to acid hydrolysis with various sulfuric acid concentrations (1-4% v/v), reaction times (1-8 h), and reaction temperatures (60-120°C). The optimal conditions for maximum total sugar production from the hydrolysate were found to be 3% sulfuric acid at 90°C for 6 h. Glucose was the predominant sugar (43.20 ± 0.28 g/L) followed by xylose (4.93 ± 0.03 g/L) and arabinose (2.09 ± 0.01 g/L). The hydrolysate was subsequently detoxified by neutralization to reduce the amount of inhibitors such as 5-hydroxymethylfurfural (HMF) and furfural to increase its potential as a medium for culturing Y. lipolytica Po1g. Dry cell mass and lipid content of Y. lipolytica Po1g grown in detoxified defatted rice bran hydrolysate (DRBH) under optimum conditions were 10.75 g/L and 48.02%, respectively. PMID:22496604

  11. Heterologous production of α-farnesene in metabolically engineered strains of Yarrowia lipolytica.

    PubMed

    Yang, Xia; Nambou, Komi; Wei, Liujing; Hua, Qiang

    2016-09-01

    Herein, we studied the heterologous production of α-farnesene, a valuable sesquiterpene with various biotechnological applications, by metabolic engineering of Yarrowia lipolytica. Different overexpression vectors harboring combinations of tHMG1, IDI, ERG20 and codon-optimized α-farnesene synthase (OptFS) genes were constructed and integrated into the genome of Y. lipolytica Po1h. The engineered strain produced 57.08±1.43mg/L of α-farnesene corresponding to 20.8-fold increase over the initial production of 2.75±0.29mg/L in the YPD medium in shake flasks. Bioreactor scale-up in PM medium led to α-farnesene concentration of 259.98±2.15mg/L with α-farnesene to biomass ratio of 33.98±1.51mg/g, which was a 94.5-fold increase over the initial production. This first report on α-farnesene synthesis in Y. lipolytica lays a foundation for future research on production of sesquitepenes in Y. lipolytica and other closest yeast species and will potentially contribute in its industrial production. PMID:27347651

  12. New Insights into Sulfur Metabolism in Yeasts as Revealed by Studies of Yarrowia lipolytica

    PubMed Central

    Hébert, Agnès; Forquin-Gomez, Marie-Pierre; Roux, Aurélie; Aubert, Julie; Junot, Christophe; Heilier, Jean-François; Landaud, Sophie; Bonnarme, Pascal

    2013-01-01

    Yarrowia lipolytica, located at the frontier of hemiascomycetous yeasts and fungi, is an excellent candidate for studies of metabolism evolution. This yeast, widely recognized for its technological applications, in particular produces volatile sulfur compounds (VSCs) that fully contribute to the flavor of smear cheese. We report here a relevant global vision of sulfur metabolism in Y. lipolytica based on a comparison between high- and low-sulfur source supplies (sulfate, methionine, or cystine) by combined approaches (transcriptomics, metabolite profiling, and VSC analysis). The strongest repression of the sulfate assimilation pathway was observed in the case of high methionine supply, together with a large accumulation of sulfur intermediates. A high sulfate supply seems to provoke considerable cellular stress via sulfite production, resulting in a decrease of the availability of the glutathione pathway's sulfur intermediates. The most limited effect was observed for the cystine supply, suggesting that the intracellular cysteine level is more controlled than that of methionine and sulfate. Using a combination of metabolomic profiling and genetic experiments, we revealed taurine and hypotaurine metabolism in yeast for the first time. On the basis of a phylogenetic study, we then demonstrated that this pathway was lost by some of the hemiascomycetous yeasts during evolution. PMID:23220962

  13. Exploring medium-chain-length polyhydroxyalkanoates production in the engineered yeast Yarrowia lipolytica.

    PubMed

    Gao, Cuijuan; Qi, Qingsheng; Madzak, Catherine; Lin, Carol Sze Ki

    2015-09-01

    Medium-chain-length polyhydroxyalkanoates (mcl-PHAs) are a large class of biopolymers that have attracted extensive attention as renewable and biodegradable bio-plastics. They are naturally synthesized via fatty acid de novo biosynthesis pathway or β-oxidation pathway from Pseudomonads. The unconventional yeast Yarrowia lipolytica has excellent lipid/fatty acid catabolism and anabolism capacity depending of the mode of culture. Nevertheless, it cannot naturally synthesize PHA, as it does not express an intrinsic PHA synthase. Here, we constructed a genetically modified strain of Y. lipolytica by heterologously expressing PhaC1 gene from P. aeruginosa PAO1 with a PTS1 peroxisomal signal. When in single copy, the codon optimized PhaC1 allowed the synthesis of 0.205 % DCW of PHA after 72 h cultivation in YNBD medium containing 0.1 % oleic acid. By using a multi-copy integration strategy, PHA content increased to 2.84 % DCW when the concentration of oleic acid in YNBD was 1.0 %. Furthermore, when the recombinant yeast was grown in the medium containing triolein, PHA accumulated up to 5.0 % DCW with as high as 21.9 g/L DCW, which represented 1.11 g/L in the culture. Our results demonstrated the potential use of Y. lipolytica as a promising microbial cell factory for PHA production using food waste, which contains lipids and other essential nutrients. PMID:26153503

  14. De novo synthesis of trans-10, cis-12 conjugated linoleic acid in oleaginous yeast Yarrowia Lipolytica

    PubMed Central

    2012-01-01

    Background Conjugated linoleic acid (CLA) has many well-documented beneficial physiological effects. Due to the insufficient natural supply of CLA and low specificity of chemically produced CLA, an effective and isomer-specific production process is required for medicinal and nutritional purposes. Results The linoleic acid isomerase gene from Propionibacterium acnes was expressed in Yarrowia lipolytica Polh. Codon usage optimization of the PAI and multi-copy integration significantly improved the expression level of PAI in Y. lipolytica. The percentage of trans-10, cis-12 CLA was six times higher in yeast carrying the codon-optimized gene than in yeast carrying the native gene. In combination with multi-copy integration, the production yield was raised to approximately 30-fold. The amount of trans-10, cis-12 CLA reached 5.9% of total fatty acid yield in transformed Y. lipolytica. Conclusions This is the first report of production of trans-10, cis-12 CLA by the oleaginous yeast Y. lipolytica, using glucose as the sole carbon source through expression of linoleic acid isomerase from Propionibacterium acnes. PMID:22545818

  15. Cloning, Expression, and Biochemical Characterization of an Enantioselective Lipase, YLIP9, from Yarrowia lipolytica MSR80.

    PubMed

    Syal, Poonam; Gupta, Rani

    2015-05-01

    A novel lipase gene, ylip9, of Yarrowia lipolytica MSR80 was cloned and expressed in pEZZ18-HB101 system and was 99% identical to YLIP9 of Y. lipolytica CLIB122. It was purified using IgG-Sepharose as ZZ fused YLIP9 and had specific activity of 0.8 U/mg. ZZ-YLIP9 was most active at pH 8.0 and 70 °C. It was stable over a wide pH range of 3.0-11.0 and 100 % active at 70 °C up to 2 h and had t1/2 of 286.42 min at 80 °C. It showed high specificity toward p-nitrophenyldecanoate with kcat and catalytic efficiency of 30.17 s(-1) and 16.67 mM(-1) s(-1), respectively. It was non-regioselective, but an S-enantioselective lipase and the percentage conversion were enhanced in presence of hexane. ZZ-YLIP9 was stable in all of the organic solvents used, and its activity was enhanced by solvents having logP value less than 2. PMID:25805014

  16. Overexpression of diacylglycerol acyltransferase in Yarrowia lipolytica affects lipid body size, number and distribution.

    PubMed

    Gajdoš, Peter; Ledesma-Amaro, Rodrigo; Nicaud, Jean-Marc; Čertík, Milan; Rossignol, Tristan

    2016-09-01

    In the oleaginous yeast Yarrowia lipolytica, the diacylglycerol acyltransferases (DGATs) are major factors for triacylglycerol (TAG) synthesis. The Q4 strain, in which the four acyltransferases have been deleted, is unable to accumulate lipids and to form lipid bodies (LBs). However, the expression of a single acyltransferase in this strain restores TAG accumulation and LB formation. Using this system, it becomes possible to characterize the activity and specificity of an individual DGAT. Here, we examined the effects of DGAT overexpression on lipid accumulation and LB formation in Y. lipolytica Specifically, we evaluated the consequences of introducing one or two copies of the Y. lipolytica DGAT genes YlDGA1 and YlDGA2 Overall, multi-copy DGAT overexpression increased the lipid content of yeast cells. However, the size and distribution of LBs depended on the specific DGAT overexpressed. YlDGA2 overexpression caused the formation of large LBs, while YlDGA1 overexpression generated smaller but more numerous LBs. This phenotype was accentuated through the addition of a second copy of the overexpressed gene and might be linked to the distinct subcellular localization of each DGAT, i.e. YlDga1 being localized in LBs, while YlDga2 being localized in a structure strongly resembling the endoplasmic reticulum. PMID:27506614

  17. Sustainable source of omega-3 eicosapentaenoic acid from metabolically engineered Yarrowia lipolytica: from fundamental research to commercial production.

    PubMed

    Xie, Dongming; Jackson, Ethel N; Zhu, Quinn

    2015-02-01

    The omega-3 fatty acids, cis-5, 8, 11, 14, and 17-eicosapentaenoic acid (C20:5; EPA) and cis-4, 7, 10, 13, 16, and 19-docosahexaenoic acid (C22:6; DHA), have wide-ranging benefits in improving heart health, immune function, mental health, and infant cognitive development. Currently, the major source for EPA and DHA is from fish oil, and a minor source of DHA is from microalgae. With the increased demand for EPA and DHA, DuPont has developed a clean and sustainable source of the omega-3 fatty acid EPA through fermentation using metabolically engineered strains of Yarrowia lipolytica. In this mini-review, we will focus on DuPont's technology for EPA production. Specifically, EPA biosynthetic and supporting pathways have been introduced into the oleaginous yeast to synthesize and accumulate EPA under fermentation conditions. This Yarrowia platform can also produce tailored omega-3 (EPA, DHA) and/or omega-6 (ARA, GLA) fatty acid mixtures in the cellular lipid profiles. Fundamental research such as metabolic engineering for strain construction, high-throughput screening for strain selection, fermentation process development, and process scale-up were all needed to achieve the high levels of EPA titer, rate, and yield required for commercial application. Here, we summarize how we have combined the fundamental bioscience and the industrial engineering skills to achieve large-scale production of Yarrowia biomass containing high amounts of EPA, which led to two commercial products, New Harvest™ EPA oil and Verlasso® salmon. PMID:25567511

  18. Strahlungsfeldbedingungen bei der Ionisationsdosimetrie

    NASA Astrophysics Data System (ADS)

    Krieger, Hanno

    Bei der Dosimetrie von Photonenstrahlungen mit luftgefüllten Ionisationskammern werden in diesem Kapitel zwei Grenzfälle unterschieden. Der eine Fall ist die Bedingung des so genannten Sekundärelektronengleichwichts im Kammervolumen. In diesem Fall bestimmen ausschließlich die Photonen im Messvolumen die dosimetrische Anzeige. Der zweite Grenzfall ist gegeben, wenn das Luftvolumen und die Kammer das Strahlungsfeld im Phantom so wenig stören, dass der Sekundärelektronenfluss unverändert bleibt. Diese Bedingungen nennt man BRAGG-GRAY-Bedingungen. Beide Grenzfälle sind in der Praxis nur näherungsweise zu verwirklichen. Abweichungen müssen durch entsprechende Korrekturen oder Kalibrierungen berücksichtigt werden.

  19. Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increased interest in sustainable production of renewable diesel and other valuable bioproducts is redoubling efforts to improve economic feasibility of microbial-based oil production. The yeast Yarrowia lipolytica is capable of employing a wide variety of substrates to produce oil and valuable co-p...

  20. The effect of yeast Yarrowia lipolytica on the antioxidant indices and macro-and microelements in blood plasma of turkey hens.

    PubMed

    Merska, M; Czech, A; Ognik, K

    2015-01-01

    The aim of the study was to assess the effect of different amounts of Yarrowia lipolytica yeast on the redox response and content of macro- and microelements in the blood plasma of turkey hens. The experiment was carried out on 240 turkey hens aged from 1 to 16 weeks. The birds were randomly assigned to 3 experimental groups of 80 birds each. Group I served as a control (K) and did not receive any experimental compounds. The turkey hens from the experimental groups (YL3 and YL6) were administered dried Yarrowia lipolytica yeast in their feed mixtures in the amount of 3% (YL3) or 6% (YL6). Yarrowia lipolytica yeast in the feed mixtures for the turkey hens did not induce oxidation reactions in the organism of the birds. However, an increase in catalase activity and a reduction in the level of LOOH, MDA and vitamin C were observed in the blood plasma of the turkey hens whose diet was supplemented with YL yeast. In the case of other indices, such as superoxide dismutase activity and total antioxidant potential (FRAP), the additive caused no significant changes. Administering Yarrowia lipolytica yeast to turkey hens may stimulate the enzymatic response of the antioxidant system (e.g. increasing catalase activity), mainly by increasing the concentration of iron in the plasma. PMID:26812811

  1. Theoretische Konzepte der Physik

    NASA Astrophysics Data System (ADS)

    Longair, Malcolm S.; Simon, B.; Simon, H.

    "Dies ist kein Lehrbuch der theoretischen Physik, auch kein Kompendium der Physikgeschichte ... , vielmehr eine recht anspruchsvolle Sammlung historischer Miniaturen zur Vergangenheit der theoretischen Physik - ihrer "Sternstunden", wenn man so will. Frei vom Zwang, etwas Erschöpfendes vorlegen zu müssen, gelingt dem Autor etwas Seltenes: einen "lebendigen" Zugang zum Ideengebäude der modernen Physik freizulegen, ... zu zeigen, wie Physik in praxi entsteht... Als Vehikel seiner Absichten dienen dem Autor geschichtliche Fallstudien, insgesamt sieben an der Zahl. Aus ihnen extrahiert er das seiner Meinung nach Lehrhafte, dabei bestrebt, mathematische Anachronismen womöglich zu vermeiden... Als Student hätte ich mir diese gescheiten Essays zum Werden unserer heutigen physikalischen Weltsicht gewünscht. Sie sind originell, didaktisch klug und genieren sich auch nicht, von der Faszination zu sprechen, die ... von der Physik ausgeht. Unnötig darauf hinzuweisen, das sie ein gründliches "konventionelles" Studium weder ersetzen wollen noch können, sie vermögen aber, dazu zu ermuntern." #Astronomische Nachrichten (zur englischen Ausgabe)#1

  2. The evolution of Jen3 proteins and their role in dicarboxylic acid transport in Yarrowia

    PubMed Central

    Dulermo, Rémi; Gamboa-Meléndez, Heber; Michely, Stéphanie; Thevenieau, France; Neuvéglise, Cécile; Nicaud, Jean-Marc

    2015-01-01

    Jen proteins in yeast are involved in the uptake of mono/dicarboxylic acids. The Jen1 subfamily transports lactate and pyruvate, while the Jen2 subfamily transports fumarate, malate, and succinate. Yarrowia lipolytica has six JEN genes: YALI0B19470g, YALI0C15488g, YALI0C21406g, YALI0D20108g, YALI0D24607g, and YALI0E32901g. Through phylogenetic analyses, we found that these genes represent a new subfamily, Jen3 and that these three Jen subfamilies derivate from three putative ancestral genes. Reverse transcription-PCR. revealed that only four YLJEN genes are expressed and they are upregulated in the presence of lactate, pyruvate, fumarate, malate, and/or succinate, suggesting that they are able to transport these substrates. Analysis of deletion mutant strains revealed that Jen3 subfamily proteins transport fumarate, malate, and succinate. We found evidence that YALI0C15488 encodes the main transporter because its deletion was sufficient to strongly reduce or suppress growth in media containing fumarate, malate, or succinate. It appears that the other YLJEN genes play a minor role, with the exception of YALI0E32901g, which is important for malate uptake. However, the overexpression of each YLJEN gene in the sextuple-deletion mutant strain ΔYLjen1-6 revealed that all six genes are functional and have evolved to transport different substrates with varying degrees of efficacy. In addition, we found that YALI0E32901p transported succinate more efficiently in the presence of lactate or fumarate. PMID:25515252

  3. Single cell oil production on molasses by Yarrowia lipolytica strains overexpressing DGA2 in multicopy.

    PubMed

    Gajdoš, Peter; Nicaud, Jean-Marc; Rossignol, Tristan; Čertík, Milan

    2015-10-01

    Yarrowia lipolytica is a promising platform for single cell oil production. It is well-known for its metabolism oriented toward utilization of hydrophobic substrates and accumulation of storage lipids. Multiple copies of DGA2 under constitutive promoter were introduced into the Q4 strain, a quadruple mutant deleted for the four acyltransferases (Δdga1, Δdga2, Δlro1, and Δare1) to improve lipid accumulation. The Q4-DGA2 x3 strain contains three copies of DGA2. Further increase in accumulation was accomplished by blocking the β-oxidation pathway through MFE1 gene deletion yielding Q4-Δmfe DGA2 x3. In order to use molasses as a substrate for single cell oil production, sucrose utilization was established by expressing the Saccharomyces cerevisiae SUC2 gene yielding Q4-SUC2 DGA2 x3 and Q4-Δmfe SUC2 DGA2 x3. During cultivation on sucrose medium with a carbon to nitrogen ratio of 80, both strains accumulated more than 40 % of lipids, which was a 2-fold increase in lipid storage. Q4-Δmfe SUC2 DGA2 x3 accumulated more lipids than Q4-SUC2 DGA2 x3 (49 vs. 43 %) but yielded less biomass (13.7 vs. 15 g/L). When grown on 8 % (v/v) molasses, both strains accumulated more than 30 % of lipids after 3 days, while biomass yield was higher in Q4-SUC2 DGA2 x3 (16.4 vs. 14.4 g/L). Further addition of molasses at 72 h resulted in higher biomass yield, 26.6 g/L for Q4-SUC2 DGA2 x3, without modification of lipid content. This work presents genetically modified strains of Y. lipolytica as suitable tools for direct conversion of industrial molasses into value added products based on single cell oils. PMID:26078110

  4. Engineering the push and pull of lipid biosynthesis in oleaginous yeast Yarrowia lipolytica for biofuel production.

    PubMed

    Tai, Mitchell; Stephanopoulos, Gregory

    2013-01-01

    Microbial oil production by heterotrophic organisms is a promising path for the cost-effective production of biofuels from renewable resources provided high conversion yields can be achieved. To this end, we have engineered the oleaginous yeast Yarrowia lipolytica. We first established an expression platform for high expression using an intron-containing translation elongation factor-1α (TEF) promoter and showed that this expression system is capable of increasing gene expression 17-fold over the intronless TEF promoter. We then used this platform for the overexpression of diacylglycerol acyltransferase (DGA1), the final step of the triglyceride (TAG) synthesis pathway, which yielded a 4-fold increase in lipid production over control, to a lipid content of 33.8% of dry cell weight (DCW). We also show that the overexpression of acetyl-CoA carboxylase (ACC1), the first committed step of fatty acid synthesis, increased lipid content 2-fold over control, or 17.9% lipid content. Next we combined the two genes in a tandem gene construct for the simultaneous coexpression of ACC1 and DGA1, which further increased lipid content to 41.4%, demonstrating synergistic effects of ACC1+DGA1 coexpression. The lipid production characteristics of the ACC1+DGA1 transformant were explored in a 2-L bioreactor fermentation, achieving 61.7% lipid content after 120h. The overall yield and productivity were 0.195g/g and 0.143g/L/h, respectively, while the maximum yield and productivity were 0.270g/g and 0.253g/L/h during the lipid accumulation phase of the fermentation. This work demonstrates the excellent capacity for lipid production by the oleaginous yeast Y. lipolytica and the effects of metabolic engineering of two important steps of the lipid synthesis pathway, which acts to divert flux towards the lipid synthesis and creates driving force for TAG synthesis. PMID:23026119

  5. Reconstruction and In Silico Analysis of Metabolic Network for an Oleaginous Yeast, Yarrowia lipolytica

    PubMed Central

    Pan, Pengcheng; Hua, Qiang

    2012-01-01

    With the emergence of energy scarcity, the use of renewable energy sources such as biodiesel is becoming increasingly necessary. Recently, many researchers have focused their minds on Yarrowia lipolytica, a model oleaginous yeast, which can be employed to accumulate large amounts of lipids that could be further converted to biodiesel. In order to understand the metabolic characteristics of Y. lipolytica at a systems level and to examine the potential for enhanced lipid production, a genome-scale compartmentalized metabolic network was reconstructed based on a combination of genome annotation and the detailed biochemical knowledge from multiple databases such as KEGG, ENZYME and BIGG. The information about protein and reaction associations of all the organisms in KEGG and Expasy-ENZYME database was arranged into an EXCEL file that can then be regarded as a new useful database to generate other reconstructions. The generated model iYL619_PCP accounts for 619 genes, 843 metabolites and 1,142 reactions including 236 transport reactions, 125 exchange reactions and 13 spontaneous reactions. The in silico model successfully predicted the minimal media and the growing abilities on different substrates. With flux balance analysis, single gene knockouts were also simulated to predict the essential genes and partially essential genes. In addition, flux variability analysis was applied to design new mutant strains that will redirect fluxes through the network and may enhance the production of lipid. This genome-scale metabolic model of Y. lipolytica can facilitate system-level metabolic analysis as well as strain development for improving the production of biodiesels and other valuable products by Y. lipolytica and other closely related oleaginous yeasts. PMID:23236514

  6. Proteomic Analysis of Erythritol-Producing Yarrowia lipolytica from Glycerol in Response to Osmotic Pressure.

    PubMed

    Yang, Li-Bo; Dai, Xiao-Meng; Zheng, Zhi-Yong; Zhu, Li; Zhan, Xiao-Bei; Lin, Chi-Chung

    2015-07-01

    Osmotic pressure is a critical factor for erythritol production with osmophilic yeast. Protein expression patterns of an erythritol-producing yeast, Yarrowia lipolytica, were analyzed to identify differentially-expressed proteins in response to osmotic pressure. In order to analyze intracellular protein levels quantitatively, two-dimensional gel electrophoresis was performed to separate and visualize the differential expression of the intracellular proteins extracted from Y. lipolytica cultured under low (3.17 osmol/kg) and high (4.21 osmol/kg) osmotic pressures. Proteomic analyses allowed identification of 54 differentially-expressed proteins among the proteins distributed in the range of pI 3-10 and 14.4-97.4 kDa molecular mass between the osmotic stress conditions. Remarkably, the main proteins were involved in the pathway of energy, metabolism, cell rescue, and stress response. The expression of such enzymes related to protein and nucleotide biosynthesis was inhibited drastically, reflecting the growth arrest of Y. lipolytica under hyperosmotic stress. The improvement of erythritol production under high osmotic stress was due to the significant induction of a range of crucial enzymes related to polyols biosynthesis, such as transketolase and triosephosphate isomerase, and the osmotic stress responsive proteins like pyridoxine-4-dehydrogenase and the AKRs family. The polyols biosynthesis was really related to an osmotic response and a protection mechanism against hyperosmotic stress in Y. lipolytica. Additionally, the high osmotic stress could also induce other cell stress responses as with heat shock and oxidation stress responses, and these responsive proteins, such as the HSPs family, catalase T, and superoxide dismutase, also had drastically increased expression levels under hyperosmotic pressure. PMID:25737116

  7. Ylli, a non-LTR retrotransposon L1 family in the dimorphic yeast Yarrowia lipolytica.

    PubMed

    Casaregola, Serge; Neuvéglise, Cécile; Bon, Elisabeth; Gaillardin, Claude

    2002-05-01

    During the course of a random sequencing project of the genome of the dimorphic yeast Yarrowia lipolytica, we have identified sequences that were repeated in the genome and that matched the reverse transcriptase (RT) sequence of non-long terminal repeat (non-LTR) retrotransposons. Extension of sequencing on each side of this zone of homology allowed the definition of an element over 6 kb long. The conceptual translation of this sequence revealed two open reading frames (ORFs) that displayed several characteristics of non-LTR retrotransposons: a Cys-rich motif in the ORF1, an N-terminal endonuclease, a central RT, and a C-terminal zinc finger domain in the ORF2. We called this element Ylli (for Y. lipolytica LINE). A total of 19 distinct repeats carrying the 3' untranslated region (UTR) and all ending with a poly-A tail were detected. Most of them were very short, 17 being 134 bp long or less. The number of copies of Ylli was estimated to be around 100 if these short repeats are 5' truncations. No 5' UTR was clearly identified, indicating that entire and therefore active elements might be very rare in the Y. lipolytica strain tested. Ylli does not seem to have any insertion specificity. Phylogenetic analysis of the RT domain unambiguously placed Ylli within the L1 clade. It forms a monophyletic group with the Zorro non-LTR retrotransposons discovered in another dimorphic yeast Candida albicans. BLAST comparisons showed that ORF2 of Ylli is closely related to that of the slime mold Dictyostelium discoideum L1 family, TRE. PMID:11961100

  8. Comparing cellular performance of Yarrowia lipolytica during growth on glucose and glycerol in submerged cultivations

    PubMed Central

    2013-01-01

    Yarrowia lipolytica is an attractive host for sustainable bioprocesses due to its ability to utilize a variety of carbon substrates and convert them to a range of different product types (including lipids, organic acids and polyols) under specific conditions. Despite an increasing number of applications for this yeast, relatively few studies have focused on uptake and metabolism of carbon sources, and the metabolic basis for carbon flow to the different products. The focus of this work was quantification of the cellular performance of Y. lipolytica during growth on glycerol, glucose or a mixture of the two. Carbon substrate uptake rate, growth rate, oxygen utilisation (requirement and uptake rate) and polyol yields were estimated in batch cultivations at 1 litre scale. When glucose was used as the sole carbon and energy source, the growth rate was 0.24 h-1 and biomass and CO2 were the only products. Growth on glycerol proceeded at approximately 0.30 h-1, and the substrate uptake rate was 0.02 mol L-1 h-1 regardless of the starting glycerol concentration (10, 20 or 45 g L-1). Utilisation of glycerol was accompanied by higher oxygen uptake rates compared to glucose growth, indicating import of glycerol occurred initially via phosphorylation of glycerol into glycerol-3-phosphate. Based on these results it could be speculated that once oxygen limitation was reached, additional production of NADH created imbalance in the cofactor pools and the polyol formation observed could be a result of cofactor recycling to restore the balance in metabolism. PMID:24088397

  9. The Strictly Aerobic Yeast Yarrowia lipolytica Tolerates Loss of a Mitochondrial DNA-Packaging Protein

    PubMed Central

    Bakkaiova, Jana; Arata, Kosuke; Matsunobu, Miki; Ono, Bungo; Aoki, Tomoyo; Lajdova, Dana; Nebohacova, Martina; Nosek, Jozef; Miyakawa, Isamu

    2014-01-01

    Mitochondrial DNA (mtDNA) is highly compacted into DNA-protein structures termed mitochondrial nucleoids (mt-nucleoids). The key mt-nucleoid components responsible for mtDNA condensation are HMG box-containing proteins such as mammalian mitochondrial transcription factor A (TFAM) and Abf2p of the yeast Saccharomyces cerevisiae. To gain insight into the function and organization of mt-nucleoids in strictly aerobic organisms, we initiated studies of these DNA-protein structures in Yarrowia lipolytica. We identified a principal component of mt-nucleoids in this yeast and termed it YlMhb1p (Y. lipolytica mitochondrial HMG box-containing protein 1). YlMhb1p contains two putative HMG boxes contributing both to DNA binding and to its ability to compact mtDNA in vitro. Phenotypic analysis of a Δmhb1 strain lacking YlMhb1p resulted in three interesting findings. First, although the mutant exhibits clear differences in mt-nucleoids accompanied by a large decrease in the mtDNA copy number and the number of mtDNA-derived transcripts, its respiratory characteristics and growth under most of the conditions tested are indistinguishable from those of the wild-type strain. Second, our results indicate that a potential imbalance between subunits of the respiratory chain encoded separately by nuclear DNA and mtDNA is prevented at a (post)translational level. Third, we found that mtDNA in the Δmhb1 strain is more prone to mutations, indicating that mtHMG box-containing proteins protect the mitochondrial genome against mutagenic events. PMID:24972935

  10. Physiological and Biochemical Responses of Yarrowia lipolytica to Dehydration Induced by Air-Drying and Freezing

    PubMed Central

    Pénicaud, Caroline; Landaud, Sophie; Jamme, Frédéric; Talbot, Pauline; Bouix, Marielle; Ghorbal, Sarrah; Fonseca, Fernanda

    2014-01-01

    Organisms that can withstand anhydrobiosis possess the unique ability to temporarily and reversibly suspend their metabolism for the periods when they live in a dehydrated state. However, the mechanisms underlying the cell’s ability to tolerate dehydration are far from being fully understood. The objective of this study was to highlight, for the first time, the cellular damage to Yarrowia lipolytica as a result of dehydration induced by drying/rehydration and freezing/thawing. Cellular response was evaluated through cell cultivability determined by plate counts, esterase activity and membrane integrity assessed by flow cytometry, and the biochemical composition of cells as determined by FT-IR spectroscopy. The effects of the harvesting time (in the log or stationary phase) and of the addition of a protective molecule, trehalose, were investigated. All freshly harvested cells exhibited esterase activity and no alteration of membrane integrity. Cells freshly harvested in the stationary phase presented spectral contributions suggesting lower nucleic acid content and thicker cell walls, as well as longer lipid chains than cells harvested in the log phase. Moreover, it was found that drying/rehydration induced cell plasma membrane permeabilization, loss of esterase activity with concomitant protein denaturation, wall damage and oxidation of nucleic acids. Plasma membrane permeabilization and loss of esterase activity could be reduced by harvesting in the stationary phase and/or with trehalose addition. Protein denaturation and wall damage could be reduced by harvesting in the stationary phase. In addition, it was shown that measurements of loss of membrane integrity and preservation of esterase activity were suitable indicators of loss and preservation of cultivability, respectively. Conversely, no clear effect of freezing/thawing could be observed, probably because of the favorable operating conditions applied. These results give insights into Y. lipolytica

  11. Sequence Assembly of Yarrowia lipolytica Strain W29/CLIB89 Shows Transposable Element Diversity.

    PubMed

    Magnan, Christophe; Yu, James; Chang, Ivan; Jahn, Ethan; Kanomata, Yuzo; Wu, Jenny; Zeller, Michael; Oakes, Melanie; Baldi, Pierre; Sandmeyer, Suzanne

    2016-01-01

    Yarrowia lipolytica, an oleaginous yeast, is capable of accumulating significant cellular mass in lipid making it an important source of biosustainable hydrocarbon-based chemicals. In spite of a similar number of protein-coding genes to that in other Hemiascomycetes, the Y. lipolytica genome is almost double that of model yeasts. Despite its economic importance and several distinct strains in common use, an independent genome assembly exists for only one strain. We report here a de novo annotated assembly of the chromosomal genome of an industrially-relevant strain, W29/CLIB89, determined by hybrid next-generation sequencing. For the first time, each Y. lipolytica chromosome is represented by a single contig. The telomeric rDNA repeats were localized by Irys long-range genome mapping and one complete copy of the rDNA sequence is reported. Two large structural variants and retroelement differences with reference strain CLIB122 including a full-length, novel Ty3/Gypsy long terminal repeat (LTR) retrotransposon and multiple LTR-like sequences are described. Strikingly, several of these are adjacent to RNA polymerase III-transcribed genes, which are almost double in number in Y. lipolytica compared to other Hemiascomycetes. In addition to previously-reported dimeric RNA polymerase III-transcribed genes, tRNA pseudogenes were identified. Multiple full-length and truncated LINE elements are also present. Therefore, although identified transposons do not constitute a significant fraction of the Y. lipolytica genome, they could have played an active role in its evolution. Differences between the sequence of this strain and of the existing reference strain underscore the utility of an additional independent genome assembly for this economically important organism. PMID:27603307

  12. Intracellular precursors and secretion of alkaline extracellular protease of Yarrowia lipolytica.

    PubMed Central

    Matoba, S; Fukayama, J; Wing, R A; Ogrydziak, D M

    1988-01-01

    Processing and secretion of the alkaline extracellular protease (AEP) from the yeast Yarrowia lipolytica was studied by pulse-chase and immunoprecipitation experiments. Over half of newly synthesized AEP was secreted by 6 min. Over 99% of AEP activity which was external to the cytoplasmic membrane was located in the supernatant medium. Polypeptides of 55, 52, 44, 36, and 32 kilodaltons (55K, 52K, 44K, 36K, and 32K polypeptides) were immunoprecipitated from [3H]leucine-labeled cell extracts by rabbit antibodies raised against mature, secreted AEP (32K polypeptide). Experiments with tunicamycin and endoglycosidase H indicated that the 55K, 52K, and 44K polypeptides contained about 2 kilodaltons of N-linked oligosaccharide and that the 36K and 32K polypeptides contained none. Results of pulse-chase experiments did not fit a simple precursor-product relationship of 55K----52K----44K----36K----32K. In fact, maximum labeling intensity of the 52K polypeptide occurred later than for the 44K and 36K polypeptides. Secretion of polypeptides of 19 and 20 kilodaltons derived from the proregion of AEP indicated that one major processing pathway was 55K----52K----32K. The gene coding for AEP (XPR2) was cloned and sequenced. The sequence and the immunoprecipitation results suggest that AEP is originally synthesized with an additional preproI-proII-proIII amino-terminal region. Processing definitely involves cleavage(s) after pairs of basic amino acids and the addition of one N-linked oligosaccharide. Signal peptidase cleavage, dipeptidyl aminopeptidase cleavages, and at least one additional proteolytic cleavage may also be involved. Images PMID:3211132

  13. Yarrowia lipolytica NCIM 3589, a tropical marine yeast, degrades bromoalkanes by an initial hydrolytic dehalogenation step.

    PubMed

    Vatsal, Aakanksha; Zinjarde, Smita S; Kumar, Ameeta Ravi

    2015-04-01

    The widespread industrial use of organobromines which are known persistent organic pollutants has led to their accumulation in sediments and water bodies causing harm to animals and humans. While degradation of organochlorines by bacteria is well documented, information regarding degradation pathways of these recalcitrant organobromines is scarce. Hence, their fates and effects on the environment are of concern. The present study shows that a tropical marine yeast, Yarrowia lipolytica NCIM 3589 aerobically degrades bromoalkanes differing in carbon chain length and position of halogen substitution viz., 2-bromopropane (2-BP), 1-bromobutane (1-BB), 1,5 dibromopentane (1,5-DBP) and 1-bromodecane (1-BD) as seen by an increase in cell mass, release of bromide and concomitant decrease in concentration of brominated compound. The amount of bromoalkane degraded was 27.3, 21.9, 18.0 and 38.3 % with degradation rates of 0.076, 0.058, 0.046 and 0.117/day for 2-BP, 1-BB, 1,5-DBP and 1-BD, respectively. The initial product formed respectively were alcohols viz., 2-propanol, 1-butanol, 1-bromo, 5-pentanol and 1-decanol as detected by GC-MS. These were further metabolized to fatty acids viz., 2-propionic, 1-butyric and 1-decanoic acid eventually leading to carbon dioxide formation. Neither higher chain nor brominated fatty acids were detected. An inducible extracellular dehalogenase responsible for removal of bromide was detected with activities of 21.07, 18.82, 18.96 and 26.67 U/ml for 2-BP, 1-BB, 1,5-DBP and 1-BD, respectively. We report here for the first time the proposed aerobic pathway of bromoalkane degradation by an eukaryotic microbe Y. lipolytica 3589, involving an initial hydrolytic dehalogenation step. PMID:25708590

  14. Nomogramme der Sickerwasserprognose

    NASA Astrophysics Data System (ADS)

    Schneider, Wilfried; Stöfen, Heinke

    Kurzfassung Modelle zur Sickerwasserprognose stehen in unterschiedlicher Komplexität zur Verfügung. Komplexe Modelle werden aufgrund der schwierigen Handhabung und des enormen Eingabedatenumfangs in der Praxis kaum angewandt. Grobe Abschätzmethoden sind dagegen nicht ausreichend wissenschaftlich fundiert, um damit justiziable Ergebnisse erzielen zu können. Um die Kluft zwischen komplexer und einfacher, jedoch justiziabler sowie wissenschaftlich fundierter Methode zu schmälern, wurden Nomogramme für Sickerwasserprognosen zur Berücksichtigung der Endlichkeit der Quelle entwickelt. Mithilfe der Nomogramme können ohne Modellierungserfahrung schnell und einfach die zu erwartenden Schadstoffkonzentrationen am Ort der Beurteilung abgeschätzt werden, falls die Endlichkeit der Quelle der hauptsächlich zur Abminderung führende Prozess ist. Die Nomogramme basieren auf analytischen Lösungen der eindimensionalen Advektions-Dispersions-Gleichung. Sie berücksichtigen die Prozesse Advektion, Diffusion in Bodenwasser und -luft, Dispersion, lineare Sorption, Abbau 1. Ordnung innerhalb einer aus mehreren Bodenschichten bestehenden Sickerwasserzone, wobei die Endlichkeit der Schadstoffmasse in der Bodenkontamination einbezogen wird. Die Genauigkeit der Nomogramme wird dargestellt. Models of different complexity are available for groundwater risk assessment. In practice complex models are hardly used, due to their difficult handling and large data requirement. Rough estimation methods are not sufficiently scientifically founded to produce justiciable results. To reduce the gap between complex and easy to use but justiciable and scientifically founded methods we developed nomograms for groundwater risk assessment which take into account the finite mass of contaminant in the source. With the help of the nomograms the expected concentrations at the point of compliance (transition between the unsaturated and saturated zone) can be estimated easily, fast and without any

  15. Immobilization of Yarrowia lipolytica lipase Ylip2 for the biocatalytic synthesis of phytosterol ester in a water activity controlled reactor.

    PubMed

    Cui, Caixia; Guan, Nan; Xing, Chen; Chen, Biqiang; Tan, Tianwei

    2016-10-01

    In this work, phytosterol ester was synthesized using Yarrowia lipolytica lipase Ylip2 that had been immobilized on inorganic support in a solvent-free system and reacted in a computer-aided water activity controlled bioreactor. The immobilization of Ylip2 on celite led to a remarkable increase in the phytosterol conversion compared to that of free lipase. An investigation of the reaction conditions were oleic acid as the fatty acid variety, 10,000U/g substrate, and a temperature of 50°C for phytosterol ester synthesis. Controlling of the water activity at a set point was accomplished by the introduction of dry air through the reaction medium at a digital feedback controlled flow rate. For the esterification of phytosterol ester, a low (15%) water activity resulted in a considerable improvement in phytosterol conversion (91.1%) as well as a decreased reaction time (78h). Furthermore, Ylip2 lipase immobilized on celite retained 90% esterification activity for the synthesis of phytosterol oleate after reused 8 cycles, while free lipase was only viable for 5 batches with 90% esterification activity remained. Finally, the phytosterol oleate space time yield increased from 1.65g/L/h with free lipase to 2.53g/L/h with immobilized lipase. These results illustrate that the immobilized Yarrowia lipolytica lipase Ylip2 in a water activity controlled reactor has great potential for the application in phytosterol esters synthesis. PMID:27416561

  16. An Alternative Approach to Synthesizing Galactooligosaccharides by Cell-Surface Display of β-Galactosidase on Yarrowia lipolytica.

    PubMed

    An, Jin; Zhang, Lebin; Li, Lijuan; Liu, Dawen; Cheng, Huiling; Wang, Hengwei; Nawaz, Muhammad Zohaib; Cheng, Hairong; Deng, Zixin

    2016-05-18

    An alternative strategy for synthesizing galactooligosaccharides (GOS) from an erythritol-producing yeast Yarrowia lipolytica using surface display technology was demonstrated. The engineered strain CGMCC11369 was developed by fusion of the β-galactosidase gene from Aspergillus oryzae to the YlPir1 gene, which codes for a cell wall protein. β-Galactosidase was effectively displayed on the cell surface of Yarrowia lipolytica start strain CGMCC7326. This engineered strain with surface-displayed β-galactosidase efficiently synthesized GOS from lactose. An amount of 160 g/L GOS was produced within 6 h in a solution of 500 g/L lactose and 5 mg/mL cell (dry weight) at pH 5.5 and 60 °C, with a yield of 51% of consumed lactose monohydrate. This newly developed method was applied with waste yeast paste from erythritol industry at least 10 times. The optimal reaction temperature increased to 60 °C, about 20 °C higher than that of free β-galactosidase, which was helpful for enhancing the reaction rate and GOS production. PMID:27090877

  17. Geologie der terrestrischen Planeten und Monde.

    NASA Astrophysics Data System (ADS)

    Stöffler, D.

    Contents: 1. Inhalt und Aufgabe der Planetologie. 2. Aufbau des Sonnensystems. 3. Methoden der Exploration und Analyse der Planeten und Monde. 4. Geologische Grundprozesse auf den terrestrischen Planeten und Monden. 5. Geologische Formationen der planetaren Oberflächen. 6. Gesteine und Mineralien der planetaren Krusten. 7. Der innere Aufbau der terrestrischen planetaren Körper. 8. Vergleichende geologische Entwicklungsgeschichte der terrestrischen Planeten und des Mondes. 9. Ursprung und Entstehung der terrestrischen planetaren Körper.

  18. Yarrowia lipolytica AAL genes are involved in peroxisomal fatty acid activation.

    PubMed

    Dulermo, Rémi; Gamboa-Meléndez, Heber; Ledesma-Amaro, Rodrigo; Thevenieau, France; Nicaud, Jean-Marc

    2016-07-01

    In yeast, β-oxidation of fatty acids (FAs) essentially takes place in peroxisomes, and FA activation must precede FA oxidation. In Saccharomyces cerevisiae, a single fatty-acyl–CoA-synthetase, ScFaa2p, mediates peroxisomal FA activation. We have previously shown that this reaction also exists in the oleaginous yeast Yarrowia lipolytica; however, the protein involved in this process remains unknown. Here, we found that proteins, named Aal proteins (Acyl/Aryl-CoA-ligases), resembling the 4-coumarate–CoA-ligase-like enzymes found in plants are involved in peroxisomal FA activation in Y. lipolytica; Y. lipolytica has 10 AAL genes, eight of which are upregulated by oleate. All the Aal proteins contain a PTS1-type peroxisomal targeting sequence (A/SKL), suggesting a peroxisomal localization. The function of the Aal proteins was analyzed using the faa1Δant1Δ mutant strain, which demonstrates neither cytoplasmic FA activation (direct result of FAA1 deletion) nor peroxisomal FA activation (indirect result of ANT1 deletion, a gene coding an ATP transporter). This strain is thus highly sensitive to external FA levels and unable to store external FAs in lipid bodies (LBs). Whereas the overexpression of (cytoplasmic) AAL1ΔPTS1 was able to partially complement the growth defect observed in the faa1Δant1Δ mutant on short-, medium- and long-chain FA media, the presence of Aal2p to Aal10p only allowed growth on the short-chain FA medium. Additionally, partial LB formation was observed in the oleate medium for strains overexpressing Aal1ΔPTS1p, Aal4ΔPTS1p, Aal7ΔPTS1p, and Aal8ΔPTS1p. Finally, an analysis of the FA content of cells grown in the oleate medium suggested that Aal4p and Aal6p present substrate specificity for C16:1 and/or C18:0. PMID:27067366

  19. Functional overexpression and characterization of lipogenesis-related genes in the oleaginous yeast Yarrowia lipolytica.

    PubMed

    Silverman, Andrew M; Qiao, Kangjian; Xu, Peng; Stephanopoulos, Gregory

    2016-04-01

    Single cell oil (SCO) is an attractive energy source due to scalability, utilization of low-cost renewable feedstocks, and type of product(s) made. Engineering strains capable of producing high lipid titers and yields is crucial to the economic viability of these processes. However, lipid synthesis in cells is a complex phenomenon subject to multiple layers of regulation, making gene target identification a challenging task. In this study, we aimed to identify genes in the oleaginous yeast Yarrowia lipolytica whose overexpression enhances lipid production by this organism. To this end, we examined the effect of the overexpression of a set of 44 native genes on lipid production in Y. lipolytica, including those involved in glycerolipid synthesis, fatty acid synthesis, central carbon metabolism, NADPH generation, regulation, and metabolite transport and characterized each resulting strain's ability to produce lipids growing on both glucose and acetate as a sole carbon source. Our results suggest that a diverse subset of genes was effective at individually influencing lipid production in Y. lipolytica, sometimes in a substrate-dependent manner. The most productive strain on glucose overexpressed the diacylglycerol acyltransferase DGA2 gene, increasing lipid titer, cellular content, and yield by 236, 165, and 246 %, respectively, over our control strain. On acetate, our most productive strain overexpressed the acylglycerol-phosphate acyltransferase SLC1 gene, with a lipid titer, cellular content, and yield increase of 99, 91, and 151 %, respectively, over the control strain. Aside from genes encoding enzymes that directly catalyze the reactions of lipid synthesis, other ways by which lipogenesis was increased in these cells include overexpressing the glycerol-3-phosphate dehydrogenase (GPD1) gene to increase production of glycerol head groups and overexpressing the 6-phosphogluconolactonase (SOL3) gene from the oxidative pentose phosphate pathway to increase NADPH

  20. Steroid biotransformations in biphasic systems with Yarrowia lipolytica expressing human liver cytochrome P450 genes

    PubMed Central

    2012-01-01

    Background Yarrowia lipolytica efficiently metabolizes and assimilates hydrophobic compounds such as n-alkanes and fatty acids. Efficient substrate uptake is enabled by naturally secreted emulsifiers and a modified cell surface hydrophobicity and protrusions formed by this yeast. We were examining the potential of recombinant Y. lipolytica as a biocatalyst for the oxidation of hardly soluble hydrophobic steroids. Furthermore, two-liquid biphasic culture systems were evaluated to increase substrate availability. While cells, together with water soluble nutrients, are maintained in the aqueous phase, substrates and most of the products are contained in a second water-immiscible organic solvent phase. Results For the first time we have co-expressed the human cytochromes P450 2D6 and 3A4 genes in Y. lipolytica together with human cytochrome P450 reductase (hCPR) or Y. lipolytica cytochrome P450 reductase (YlCPR). These whole-cell biocatalysts were used for the conversion of poorly soluble steroids in biphasic systems. Employing a biphasic system with the organic solvent and Y. lipolytica carbon source ethyl oleate for the whole-cell bioconversion of progesterone, the initial specific hydroxylation rate in a 1.5 L stirred tank bioreactor was further increased 2-fold. Furthermore, the product formation was significantly prolonged as compared to the aqueous system. Co-expression of the human CPR gene led to a 4-10-fold higher specific activity, compared to the co-overexpression of the native Y. lipolytica CPR gene. Multicopy transformants showed a 50-70-fold increase of activity as compared to single copy strains. Conclusions Alkane-assimilating yeast Y. lipolytica, coupled with the described expression strategies, demonstrated its high potential for biotransformations of hydrophobic substrates in two-liquid biphasic systems. Especially organic solvents which can be efficiently taken up and/or metabolized by the cell might enable more efficient bioconversion as compared

  1. Comprehensive Metabolomic, Lipidomic and Microscopic Profiling of Yarrowia lipolytica during Lipid Accumulation Identifies Targets for Increased Lipogenesis

    PubMed Central

    Pomraning, Kyle R.; Wei, Siwei; Karagiosis, Sue A.; Kim, Young-Mo; Dohnalkova, Alice C.; Arey, Bruce W.; Bredeweg, Erin L.; Orr, Galya; Metz, Thomas O.; Baker, Scott E.

    2015-01-01

    Yarrowia lipolytica is an oleaginous ascomycete yeast that accumulates large amounts of lipids and has potential as a biofuel producing organism. Despite a growing scientific literature focused on lipid production by Y. lipolytica, there remain significant knowledge gaps regarding the key biological processes involved. We applied a combination of metabolomic and lipidomic profiling approaches as well as microscopic techniques to identify and characterize the key pathways involved in de novo lipid accumulation from glucose in batch cultured, wild-type Y. lipolytica. We found that lipids accumulated rapidly and peaked at 48 hours during the five day experiment, concurrent with a shift in amino acid metabolism. We also report that exhaustion of extracellular sugars coincided with thickening of the cell wall, suggesting that genes involved in cell wall biogenesis may be a useful target for improving the efficiency of lipid producing yeast strains. PMID:25905710

  2. The effect of oxalic and itaconic acids on threo-Ds-isocitric acid production from rapeseed oil by Yarrowia lipolytica.

    PubMed

    Kamzolova, Svetlana V; Allayarov, Ramil K; Lunina, Julia N; Morgunov, Igor G

    2016-04-01

    The effect of oxalic and itaconic acids, the inhibitors of the isocitrate lyase, on the production of isocitric acid by the wild strain Yarrowia lipolytica VKM Y-2373 grown in the medium containing rapeseed oil was studied. In the presence of oxalic and itaconic acids, strain Y. lipolytica accumulated in the medium isocitric acid (70.0 and 82.7 g/L, respectively) and citric acid (23.0 and 18.4 g/L, respectively). In control experiment, when the inhibitors were not added to the medium, the strain accumulated isocitric and citric acids at concentrations of 62.0 and 28.0 g/L, respectively. Thus, the use of the oxalic and itaconic acids as additives to the medium is a simple and convenient method of isocitric acid production with a minimum content of citric acid. PMID:26851896

  3. Efficient conversion of crude glycerol from various industrial wastes into single cell oil by yeast Yarrowia lipolytica.

    PubMed

    Dobrowolski, Adam; Mituła, Paweł; Rymowicz, Waldemar; Mirończuk, Aleksandra M

    2016-05-01

    In this study, crude glycerol from various industries was used to produce lipids via wild type Yarrowia lipolytica A101. We tested samples without any prior purification from five different waste products; each contained various concentrations of glycerol (42-87%) as the sole carbon source. The best results for lipid production were obtained for medium containing glycerol from fat saponification. This reached 1.69gL(-1) (25% of total cell dry weight) with a biomass yield of 0.17gg(-1) in the flasks experiment. The batch cultivation in a bioreactor resulted in enhanced lipid production-it achieved 4.72gL(-1) with a biomass yield 0.21gg(-1). Moreover, the properly selected batch of crude glycerol provides a defined fatty acid composition. In summary, this paper shows that crude glycerol from soap production could be efficiently converted to single cell oil without any prior purification. PMID:26890799

  4. Surveying the lipogenesis landscape in Yarrowia lipolytica through understanding the function of a Mga2p regulatory protein mutant.

    PubMed

    Liu, Leqian; Markham, Kelly; Blazeck, John; Zhou, Nijia; Leon, Dacia; Otoupal, Peter; Alper, Hal S

    2015-09-01

    Lipogenic organisms represent great starting points for metabolic engineering of oleochemical production. While previous engineering efforts were able to significantly improve lipid production in Yarrowia lipolytica, the lipogenesis landscape, especially with respect to regulatory elements, has not been fully explored. Through a comparative genomics and transcriptomics approach, we identified and validated a mutant mga2 protein that serves as a regulator of desaturase gene expression and potent lipogenesis factor. The resulting strain is enriched in unsaturated fatty acids. Comparing the underlying mechanism of this mutant to other previously engineered strains suggests that creating an imbalance between glycolysis and the TCA cycle can serve as a driving force for lipogenesis when combined with fatty acid catabolism overexpressions. Further comparative transcriptomics analysis revealed both distinct and convergent rewiring associated with these different genotypes. Finally, by combining metabolic engineering targets, it is possible to further engineer a strain containing the mutant mga2 gene to a lipid production titer of 25g/L. PMID:26219673

  5. Comprehensive metabolomic, lipidomic and microscopic profiling of Yarrowia lipolytica during lipid accumulation identifies targets for increased lipogenesis

    SciTech Connect

    Pomraning, Kyle R.; Wei, Siwei; Karagiosis, Sue A.; Kim, Young-Mo; Dohnalkova, Alice; Arey, Bruce W.; Bredeweg, Erin L.; Orr, Galya; Metz, Thomas O.; Baker, Scott E.

    2015-04-23

    Yarrowia lipolytica is an oleaginous ascomycete yeast that accumulates large amounts of lipids and has potential as a biofuel producing organism. Despite a growing scientific literature focused on lipid production by Y. lipolytica, there remain significant knowledge gaps regarding the key biological processes involved. We applied a combination of metabolomic and lipidomic profiling approaches as well as microscopic techniques to identify and characterize the key pathways involved in de novo lipid accumulation from glucose in batch cultured, wild-type Y. lipolytica. We found that lipids accumulated rapidly and peaked at 48 hours during the five day experiment, concurrent with a shift in amino acid metabolism. We also report that Y. lipolytica secretes disaccharides early in batch culture and reabsorbs them when extracellular glucose is depleted. Exhaustion of extracellular sugars coincided with thickening of the cell wall, suggesting that genes involved in cell wall biogenesis may be a useful target for improving the efficiency of lipid producing yeast strains.

  6. Die Zeitung der Zukunft

    NASA Astrophysics Data System (ADS)

    Wieser, Christoph; Schaffert, Sebastian

    Schon lange wird spekuliert, wie wir in Zukunft Zeitung lesen werden. Werden wir am Frühstückstisch wie gewohnt in einer Zeitung aus Papier schmökern oder werden wir die Zeitung als biegsame Folie beschrieben mit elektronischer Tinte in Händen halten? Wird die Zeitung mit anderen Medien wie Radio und Fernsehen verschmelzen? Viele Varianten sind denkbar. Heute lässt sich schon ein Trend ablesen: Immer mehr Leser entdecken die Online-Zeitung als Informationsmedium, eine Voraussetzung für die Nutzung neuer Technologien in der Zeitung der Zukunft. In diesem Kapitel stellen wir Entwicklungsmöglichkeiten der Online-Zeitung dar, wie sie im Social Semantic Web möglich werden.

  7. Zum Stellenwert der Unterdruck-Instillationstherapie in der Dermatologie.

    PubMed

    Müller, Cornelia Sigrid Lissi; Burgard, Barbara; Zimmerman, Monika; Vogt, Thomas; Pföhler, Claudia

    2016-08-01

    Die Methoden zur Behandlung akuter und chronischer Wunden unterliegen einer steten Weiterentwicklung, Reevaluierung und Anwendung innovativer Therapieformen. Die Vakuumtherapie zur Wundbehandlung gehört zu den etablierten Behandlungsmodalitäten. Ein innovatives Verfahren kombiniert die Vakuumtherapie mit der automatisierten, kontrollierten Zufuhr und Drainage wirkstoffhaltiger Lösungen zur topischen Wundbehandlung im Wundbett und auch wirkstofffrei durch Instillation physiologischer Kochsalzlösung (Unterdruck-Instillationstherapie). Hierdurch können die Effekte der konventionellen Vakuumtherapie mit denen der lokalen Antisepsis kombiniert werden. Hierdurch kommt es zu einer Reduktion der Wundfläche, einer Induktion von Granulationsgewebe sowie einer Reduktion der Keimbesiedelung der Wunden. Bisher publizierte Studien konzentrieren sich auf die Anwendung dieses Therapieverfahrens zur Behandlung orthopädisch-chirurgischer Krankheiten. Die Datenlage bezüglich der Vakuum-Instillationstherapie in der Dermatochirurgie beschränkt sich derzeit auf Fallberichte und Einzelfallerfahrungen. Randomisierte, prospektive Studien zum Vergleich der Vakuum-Instillationstherapie zur Behandlung dermatologischer Krankheitsbilder existieren bislang nicht. Ziele des vorliegenden Artikels sind die Vorstellung der Vakuumtherapie mit Instillation einschließlich ihres Wirkprinzips, deren mögliche Komplikationen, die Diskussion erdenklicher Kontraindikationen sowie eine Übersicht über die aktuell verfügbare Datenlage. Zusammenfassend scheint sich die Evidenz zu verdichten, dass mittels Unterdruck-Instillationstherapie sowohl einfache als auch komplizierte Wunden effizient behandelt werden können, was sich in einer deutlichen Beschleunigung der Wundgranulation mit konsekutiv früher möglichem Defektverschluss äußert. PMID:27509413

  8. Clinical Characteristics, Laboratory Identification, and In Vitro Antifungal Susceptibility of Yarrowia (Candida) lipolytica Isolates Causing Fungemia: a Multicenter, Prospective Surveillance Study

    PubMed Central

    Zhao, Ying; Chan, Jasper Fuk-Woo; Tsang, Chi-Ching; Wang, He; Guo, Dawen; Pan, Yuhong; Xiao, Yuling; Yue, Na; Chen, Jonathan Hon-Kwan; Lau, Susanna Kar-Pui

    2015-01-01

    Our case series showed that uncomplicated Yarrowia lipolytica fungemia might be treated with catheter removal alone. The Vitek 2 YST identification (ID) card system, matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS), and internal transcribed spacer and 25S nuclear ribosomal DNA (nrDNA) gene sequencing provided reliable identification. All isolates had low MICs to voriconazole, echinocandins, and amphotericin B. PMID:26311865

  9. Clinical Characteristics, Laboratory Identification, and In Vitro Antifungal Susceptibility of Yarrowia (Candida) lipolytica Isolates Causing Fungemia: a Multicenter, Prospective Surveillance Study.

    PubMed

    Zhao, Ying; Chan, Jasper Fuk-Woo; Tsang, Chi-Ching; Wang, He; Guo, Dawen; Pan, Yuhong; Xiao, Yuling; Yue, Na; Chen, Jonathan Hon-Kwan; Lau, Susanna Kar-Pui; Xu, Yingchun; Woo, Patrick Chiu-Yat

    2015-11-01

    Our case series showed that uncomplicated Yarrowia lipolytica fungemia might be treated with catheter removal alone. The Vitek 2 YST identification (ID) card system, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and internal transcribed spacer and 25S nuclear ribosomal DNA (nrDNA) gene sequencing provided reliable identification. All isolates had low MICs to voriconazole, echinocandins, and amphotericin B. PMID:26311865

  10. Engineering Yarrowia lipolytica to Produce Glycoproteins Homogeneously Modified with the Universal Man3GlcNAc2 N-Glycan Core

    PubMed Central

    De Pourcq, Karen; Tiels, Petra; Van Hecke, Annelies; Geysens, Steven; Vervecken, Wouter; Callewaert, Nico

    2012-01-01

    Yarrowia lipolytica is a dimorphic yeast that efficiently secretes various heterologous proteins and is classified as “generally recognized as safe.” Therefore, it is an attractive protein production host. However, yeasts modify glycoproteins with non-human high mannose-type N-glycans. These structures reduce the protein half-life in vivo and can be immunogenic in man. Here, we describe how we genetically engineered N-glycan biosynthesis in Yarrowia lipolytica so that it produces Man3GlcNAc2 structures on its glycoproteins. We obtained unprecedented levels of homogeneity of this glycanstructure. This is the ideal starting point for building human-like sugars. Disruption of the ALG3 gene resulted in modification of proteins mainly with Man5GlcNAc2 and GlcMan5GlcNAc2 glycans, and to a lesser extent with Glc2Man5GlcNAc2 glycans. To avoid underoccupancy of glycosylation sites, we concomitantly overexpressed ALG6. We also explored several approaches to remove the terminal glucose residues, which hamper further humanization of N-glycosylation; overexpression of the heterodimeric Apergillus niger glucosidase II proved to be the most effective approach. Finally, we overexpressed an α-1,2-mannosidase to obtain Man3GlcNAc2 structures, which are substrates for the synthesis of complex-type glycans. The final Yarrowia lipolytica strain produces proteins glycosylated with the trimannosyl core N-glycan (Man3GlcNAc2), which is the common core of all complex-type N-glycans. All these glycans can be constructed on the obtained trimannosyl N-glycan using either in vivo or in vitro modification with the appropriate glycosyltransferases. The results demonstrate the high potential of Yarrowia lipolytica to be developed as an efficient expression system for the production of glycoproteins with humanized glycans. PMID:22768188

  11. Van der Waals Forces

    NASA Astrophysics Data System (ADS)

    Parsegian, V. Adrian

    2006-03-01

    This should prove to be the definitive work explaining van der Waals forces, how to calculate them and take account of their impact under any circumstances and conditions. These weak intermolecular forces are of truly pervasive impact, and biologists, chemists, physicists and engineers will profit greatly from the thorough grounding in these fundamental forces that this book offers. Parsegian has organized his book at three successive levels of mathematical sophistication, to satisfy the needs and interests of readers at all levels of preparation. The Prelude and Level 1 are intended to give everyone an overview in words and pictures of the modern theory of van der Waals forces. Level 2 gives the formulae and a wide range of algorithms to let readers compute the van der Waals forces under virtually any physical or physiological conditions. Level 3 offers a rigorous basic formulation of the theory. Author is among the most highly respected biophysicists Van der Waals forces are significant for a wide range of questions and problems in the life sciences, chemistry, physics, and engineering, ranging up to the macro level No other book that develops the subject vigorously, and this book also makes the subject intuitively accessible to students who had not previously been mathematically sophisticated enough to calculate them

  12. Das mechatronische Fahrwerk der Zukunft

    NASA Astrophysics Data System (ADS)

    Rieth, Peter

    Technologiesprünge gab es immer wieder in der Geschichte der Automobiltechnologie, so auch in der Kraftfahrzeugbremse, die seit ihren Anfängen eine stete Weiterentwicklung erfahren hat (Bild 40-1). So erlangten schon mechanische Bremsen durchaus ein hohes Niveau, ehe die Hydraulik Mitte der zwanziger Jahre für Komfort und Sicherheit vollkommen neue Perspektiven eröffnete, welche durch Einführung der Hilfskraft(servo)bremsanlagen Mitte des letzten Jahrhunderts noch erweitert wurden. Den wichtigsten Technologiesprung, den der Kunde als Fortschritt erkennt und dementsprechend honoriert, ermöglichte die Elektronik Mitte der siebziger Jahre. ABS, ASR, EBV und nicht zuletzt ESC wären ohne sie nicht vorstellbar.

  13. Evaluation der zentralen TUM-Lernplattform

    NASA Astrophysics Data System (ADS)

    Schulze, Elvira; Baume, Matthias; Graf, Stephan; Gergintchev, Ivan

    Die Notwendigkeit der Qualitätssicherung und -kontrolle für innovative universitäre Lehr-/Lernszenarien ist in der Praxis unbestritten. Die Wirksamkeit der Einführung der zentralen Lernplattform CLIX Campus der imc AG an der TUM wurde mittels quantitativer und qualitativer Evaluation überprüft. Als statistische Bewertungsgrundlage wurde der Erreichungsgrad bestimmter Projektziele herangezogen. Aufbauend auf den theoretischen Grundlagen der Evaluation von Bildungsangeboten gibt diese Studie Aufschluss über die Ergebnisse der Datenerhebungen sowie die Einschätzung der Plattform aus Nutzersicht und belegt die wesentliche Bedeutung der durchgängigen IT-Infrastruktur und speziell der einheitlichen Verfügbarkeit der eLearning Angebote.

  14. Haftung in der Medizintechnik

    NASA Astrophysics Data System (ADS)

    Müller, Ute; Lücker, Volker

    Die Unversehrtheit von Leib und Leben ist das größte Rechtsgut unserer Gesellschaft. Dies macht schon das Grundgesetz in Art. 2 Abs. Satz 1 GG deutlich. Die Öffentlichkeit zeigt daher größtes Interesse an Produkten, welche der Gesundheit dienen und Leben retten oder erhalten. Dieses Interesse gilt einerseits der Entwicklung und Bereitstellung leistungsfähiger Medizinprodukte, andererseits zielt es auf deren Sicherheit. Um vor allem letztere zu gewährleisten, nimmt der Gesetzgeber alle Beteiligten in die Pflicht, die auftretenden Risiken auf das geringstmögliche Maß zu begrenzen. Dies spiegelt sich in den rechtlichen Vorgaben ebenso wie in den Haftungsfolgen, die bei Verletzung dieser Vorgaben greifen, wieder. Diese Folgen können dementsprechend gravierend ausfallen, von Geldstrafen bis zu Freiheitsstrafen, von Bußgeldzahlungen bis zum Schadenersatzansprüchen, die schnell ein wirtschaftliches Aus bedeuten können. Den Beteiligten, allen voran den Herstellern, muss deshalb daran gelegen sein, nicht nur die Produkte, sondern auch deren Sicherheit stetig weiter zu entwickeln.

  15. Der Forsch-Frosch Fred

    NASA Astrophysics Data System (ADS)

    Kaibel, Volker

    Bewegt man sich auf der Suche nach Informationen zu einem bestimmten Thema immer neuen Links folgend durch das World-Wide-Web, so stellt man in der Regel nach einer gewissen Zeit fest, dass man auf manchen Seiten immer wieder gelandet ist. Diese Seiten stellen sich oft als wichtig heraus. Suchmaschinen wie Google machen sich dieses Phänomen zu Nutze, um die Wichtigkeit einzelner Web-Seiten zu bewerten. Dazu lassen sie auf einer beliebigen Web-Seite einen imaginären Surfer starten, der immer zufällig einen der auf seiner aktuell besuchten Seite vorgefundenen Links verfolgt. Der Surfer vollführt einen Random Walk im Netzwerk der Web-Seiten.

  16. A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass.

    PubMed

    Kalyani, Dayanand; Tiwari, Manish Kumar; Li, Jinglin; Kim, Sun Chang; Kalia, Vipin C; Kang, Yun Chan; Lee, Jung-Kul

    2015-01-01

    A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification. PMID:25781945

  17. Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica

    PubMed Central

    Nambou, Komi; Jian, Xingxing; Zhang, Xinkai; Wei, Liujing; Lou, Jiajia; Madzak, Catherine; Hua, Qiang

    2015-01-01

    Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA) and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species. PMID:26703753

  18. Simultaneous saccharification and fermentation of cellulose in ionic liquid for efficient production of α-ketoglutaric acid by Yarrowia lipolytica.

    PubMed

    Ryu, Seunghyun; Labbé, Nicole; Trinh, Cong T

    2015-05-01

    Ionic liquids (ILs) are benign solvents that are highly effective for biomass pretreatment. However, their applications for scale-up biorefinery are limited due to multiple expensive IL recovery and separation steps that are required. To overcome this limitation, it is very critical to develop a compatible enzymatic and microbial biocatalyst system to carry the simultaneous saccharification and fermentation in IL environments (SSF-IL). While enzymatic biocatalysts have been demonstrated to be compatible with various IL environments, it is challenging to develop microbial biocatalysts that can thrive and perform efficient biotransformation under the same conditions (pH and temperature). In this study, we harnessed the robust metabolism of Yarrowia lipolytica as a microbial platform highly compatible with the IL environments such as 1-ethyl-3-methylimidazolium acetate ([EMIM][OAc]). We optimized the enzymatic and microbial biocatalyst system using commercial cellulases and demonstrated the capability of Y. lipolytica to convert cellulose into high-value organics such as α-ketoglutaric acid (KGA) in the SSF-IL process at relatively low temperature 28 °C and high pH 6.3. We showed that SSF-IL not only enhanced the enzymatic saccharification but also produced KGA up to 92% of the maximum theoretical yield. PMID:25783627

  19. Intracellular expression of Vitreoscilla hemoglobin improves production of Yarrowia lipolytica lipase LIP2 in a recombinant Pichia pastoris.

    PubMed

    Wang, Xiaofeng; Sun, Yongchuan; Shen, Xuguang; Ke, Feng; Zhao, Heyun; Liu, Yun; Xu, Li; Yan, Yunjun

    2012-01-01

    The Yarrowia lipolytica lipase LIP2 (YlLIP2) gene lip2 and Vitreoscilla hemoglobin gene vgb were co-expressed in Pichia pastoris, both under the control of AOX1 promoter, in order to alleviate respiration limitation under conditions of high cell-density fermentation and enhance YlLIP2 production. The results showed that recombinant P. pastoris strains harboring the lip2 and vgb genes (VHb(+)) displayed higher biomass and YlLIP2 activity than control strains (VHb(-)). Compared with VHb(-) cells, the expression levels of YlLIP2 in VHb-expressing cells when oxygen was not a limiting factor were improved 31.5% in shake-flask culture and 22% in a 10-L fermentor. Under non-limiting dissolved oxygen (DO) conditions, the maximum YlLIP2 activity of VHb(+) in a 10-L fermentor reached 33,000 U/mL. Oxygen limitation had a more negative effect on YlLIP2 productivity in VHb(-) cells than in VHb(+) cells. The highest YlLIP2 activity of VHb(+) cells was approximately 1.84-fold higher than that of VHb(-) cells at lower DO levels. Moreover, the recombinant strain VHb(+) exhibited a higher specific oxygen uptake rate and achieved higher cell viability under oxygen limiting and non-limiting conditions compared with VHb(-) cells. Therefore, the above results suggest that intracellular expression of VHb in recombinant P. pastoris has the potential to improve cell growth and industrial enzyme production. PMID:22133436

  20. An enhanced process for the production of a highly purified extracellular lipase in the non-conventional yeast Yarrowia lipolytica.

    PubMed

    Turki, Saoussen; Ayed, Atef; Chalghoumi, Néjib; Weekers, Frederic; Thonart, Philippe; Kallel, Héla

    2010-03-01

    Yarrowia lipolytica LgX64.81 is a non-genetically modified mutant that was previously identified as a promising microorganism for extracellular lipase production. In this work, the development of a fed-batch process for the production of this enzyme in this strain was described. A lipolytic activity of 2,145 U/mL was obtained after 32 h of batch culture in a defined medium supplemented with 10 g/L of tryptone, an enhancer of lipase expression. To maximize the volumetric productivity, two different fed-batch strategies had been investigated. In comparison to batch process, the intermittent fed-batch strategy had not improved the volumetric lipase productivity. In contrast, the stepwise feeding strategy combined with uncoupled cell growth and lipase production phases resulted in a 2-fold increase in the volumetric lipase productivity, namely, the lipase activity reached 10,000 U/mL after 80 h of culture. Furthermore, this lipase was purified to homogeneity by anion exchange chromatography on MonoQ resin followed by gel filtration on Sephacryl S-100. This process resulted in an overall yield of 72% and a 3.5-fold increase of the specific lipase activity. The developed process offers a great potential for an economic production of Lip2 at large scale in Y. lipolytica LgX64.81. PMID:19333561

  1. Probing role of key residues in the divergent evolution of Yarrowia lipolytica lipase 2 and Aspergillus niger eruloyl esterase A.

    PubMed

    Wang, Guilong; Liu, Zimin; Xu, Li; Zhang, Houjin; Yan, Yunjun

    2015-09-01

    Yarrowia lipolytica lipase 2 (YLLip2) and Aspergillus niger feruloyl esterase A (AnFaeA) are enzymes of similar structures but with different functions. They are both classified into the same homologous family in Lipase Engineering Database (LED). The major difference between the two enzymes is that YLLip2 exhibits interfacial activity while AnFaeA does not. In order to better understand the interfacial activation mechanisms of YLLip2, structure guided site-directed mutagenesis were performed, mutants were constructed, kinetics parameters and lipase properties were detected. Mutant enzymes showed enhanced catalytic efficiency towards p-nitrophenyl butyrin (pNPB) but their catalytic efficiency decreased towards p-nitrophenyl palmitate (pNPP), their catalysis behavior was more close to feruloyl esterase. Moreover, the mutant enzymes exhibited enhanced thermostability compared with their wild type. These results indicate that I100 and F129 are probably cut-off point of divergent functions between the two enzymes during evolution. PMID:26302844

  2. Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host.

    PubMed

    Celińska, Ewelina; Białas, Wojciech; Borkowska, Monika; Grajek, Włodzimierz

    2015-03-01

    Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, α-amylase from rice weevil (Sitophilus oryzae), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81 AU/L) was reached after only 29 h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5 % yield). The optimal conditions for the α-amylase activity were pH 5.0 and a temperature of 40 °C. The α-amylase studied here did not show any obligate requirement for Ca(2+) ions. The recombinant α-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice. PMID:25547839

  3. Optimization of biomass production of a mutant of Yarrowia lipolytica with an increased lipase activity using raw glycerol.

    PubMed

    Galvagno, Miguel A; Iannone, Leopoldo J; Bianchi, Jorgelina; Kronberg, Florencia; Rost, Enrique; Carstens, Maria R; Cerrutti, Patricia

    2011-01-01

    The yeast Yarrowia lipolytica accumulates oils and is able to produce extracellular lipases when growing in different carbon sources including glycerol, the principal by-product of the biodiesel industry. In this study, biomass production of a novel mutant strain of Y. lipolytica was statistically optimized by Response Surface Methodology in media containing biodiesel-derived glycerol as main carbon source. This strain exhibited distinctive morphological and fatty acid profile characteristics, and showed an increased extracellular lipase activity. An organic source of nitrogen and the addition of 1.0 g/l olive oil were necessary for significant lipase production. Plackett-Burman and Central Composite Statistical Designs were employed for screening and optimization of fermentation in shaken flasks cultures, and the maximum values obtained were 16.1 g/l for biomass and 12.2 Units/ml for lipase, respectively. Optimized batch bioprocess was thereafter scaled in aerated bioreactors and the values reached for lipase specific activity after 95 % of the glycerol had been consumed, were three-fold higher than those obtained in shaken flasks cultures. A sustainable bioprocess to obtain biomass and extracellular lipase activity was attained by maximizing the use of the by-products of biodiesel industry. PMID:22430997

  4. Enhanced α-ketoglutaric acid production and recovery in Yarrowia lipolytica yeast by effective pH controlling.

    PubMed

    Morgunov, Igor G; Kamzolova, Svetlana V; Samoilenko, Vladimir A

    2013-10-01

    The replacement of chemical synthesis by environmentally friendly energy-efficient technologies for production of valuable metabolites is a principal strategy of developing biotechnological industry all over the world. In the present study, we develop a method for α-ketoglutaric acid (KGA) production from rapeseed oil with the use of Yarrowia lipolytica yeast. Sixty strains of Y. lipolytica yeasts were tested for their ability to produce KGA, and the strain Y. lipolytica 212 (Y. lipolytica VKM Y-2412) was selected as a promising KGA producer. Using a three-stage pH controlling, in which pH was 4.5 in the growth phase, then since 72 to 144 h, pH was maintained at 3.5 and in the later phase of acid production, the titration by KOH was switch off, selected strain produced 106.5 g l(-1) of KGA with mass yield of 0.95 g g(-1). KGA in the form of monopotassium salt was isolated from the culture broth and purified. The isolation procedure involved separation of biomass, extraction of residual triglycerides, filtrate bleaching, and acidification with mineral acid (to pH 2.8-3.4), concentration, precipitation of mineral salts, and crystallization of the product. The purity of KGA isolated from the culture filtrate reached 99.1 %. PMID:23948727

  5. Immobilization of Yarrowia lipolytica Lipase—A Comparison of Stability of Physical Adsorption and Covalent Attachment Techniques

    NASA Astrophysics Data System (ADS)

    Cunha, Aline G.; Fernández-Lorente, Gloria; Bevilaqua, Juliana V.; Destain, Jacqueline; Paiva, Lúcia M. C.; Freire, Denise M. G.; Fernández-Lafuente, Roberto; Guisán, Jose M.

    Lipase immobilization offers unique advantages in terms of better process control, enhanced stability, predictable decay rates and improved economics. This work evaluated the immobilization of a highly active Yarrowia lipolytica lipase (YLL) by physical adsorption and covalent attachment. The enzyme was adsorbed on octyl-agarose and octadecyl-sepabeads supports by hydrophobic adsorption at low ionic strength and on MANAE-agarose support by ionic adsorption. CNBr-agarose was used as support for the covalent attachment immobilization. Immobilization yields of 71, 90 and 97% were obtained when Y. lipolytica lipase was immobilized into octyl-agarose, octadecyl-sepabeads and MANAE-agarose, respectively. However, the activity retention was lower (34% for octyl-agarose, 50% for octadecyl-sepabeads and 61% for MANAE-agarose), indicating that the immobilized lipase lost activity during immobilization procedures. Furthermore, immobilization by covalent attachment led to complete enzyme inactivation. Thermal deactivation was studied at a temperature range from 25 to 45°C and pH varying from 5.0 to 9.0 and revealed that the hydrophobic adsorption on octadecyl-sepabeads produced an appreciable stabilization of the biocatalyst. The octadecyl-sepabeads biocatalyst was almost tenfold more stable than free lipase, and its thermal deactivation profile was also modified. On the other hand, the Y. lipolytica lipase immobilized on octyl-agarose and MANAE-agarose supports presented low stability, even less than the free enzyme.

  6. Physiologo-biochemical characteristics of citrate-producing yeast Yarrowia lipolytica grown on glycerol-containing waste of biodiesel industry.

    PubMed

    Morgunov, Igor G; Kamzolova, Svetlana V

    2015-08-01

    In this study, physiologo-biochemical characteristics of citrate-producing yeast Yarrowia lipolytica grown on glycerol-containing waste of biodiesel industry were studied by an investigation of growth dynamics, the consumption of glycerol, and the fatty acid fractions from waste as well as by measuring the activities of enzymes involved in the metabolism of waste. It was shown that Y. lipolytica realizes concurrent uptake of glycerol and the fatty acid fractions during conversion of glycerol-containing waste, although glycerol was utilized at a higher rate than fatty acids. Under optimal feeding of glycerol-containing waste by portions of 20 g l(-1), the citric acid production and the ratio between citric acid and isocitric acid depended on the strain used. It was revealed that wild strain Y. lipolytica VKM Y-2373 produced citrate and isocitrate with a ratio of 1.7:1, while the mutant strain Y. lipolytica NG40/UV7 synthesized presumably citric acid (122.2 g l(-1)) with a citrate-to-isocitrate ratio of 53:1 and the yield of 0.95 g g(-1). PMID:25846335

  7. Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica.

    PubMed

    Nambou, Komi; Jian, Xingxing; Zhang, Xinkai; Wei, Liujing; Lou, Jiajia; Madzak, Catherine; Hua, Qiang

    2015-01-01

    Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA) and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species. PMID:26703753

  8. Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica

    PubMed Central

    2012-01-01

    In this study, a novel rDNA based plasmid was developed for display of heterologous proteins on the cell surface of Yarrowia lipolytica using the C-terminal end of the glycosylphosphatidylinositol (GPI) anchored Y. lipolytica cell wall protein 1 (YlCWP1). mCherry was used as a model protein to assess the efficiency of the constructed plasmid. Y. lipolytica transformants harbouring the expression cassettes showed a purple colour phenotype on selective YNB-casamino plates as compared to control cells indicating that mCherry was displayed on the cells. Expression of mCherry on cells of Y. lipolytica was confirmed by both fluorescent microscopy and flow cytometry. Furthermore, SDS-PAGE analysis and matrix-assisted laser desorption/ionization (MALDI)-time-of (TOF)-mass spectrometry (MS) peptide mass fingerprinting (PMF) confirmed that the protein cleaved from the yeast cells using enterokinase was mCherry. Efficient cleavage of mCherry reported in this work offers an alternative purification method for displayed heterologous proteins on Y. lipolytica cells using the plasmid constructed in this study. The developed displaying system offers great potential for industrial production and purification of heterologous proteins at low cost. PMID:22608131

  9. The expression of the Cuphea palustris thioesterase CpFatB2 in Yarrowia lipolytica triggers oleic acid accumulation.

    PubMed

    Stefan, Alessandra; Hochkoeppler, Alejandro; Ugolini, Luisa; Lazzeri, Luca; Conte, Emanuele

    2016-01-01

    The conversion of industrial by-products into high-value added compounds is a challenging issue. Crude glycerol, a by-product of the biodiesel production chain, could represent an alternative carbon source for the cultivation of oleaginous yeasts. Here, we developed five minimal synthetic glycerol-based media, with different C/N ratios, and we analyzed the production of biomass and fatty acids by Yarrowia lipolytica Po1g strain. We identified two media at the expense of which Y. lipolytica was able to accumulate ∼5 g L(-1) of biomass and 0.8 g L(-1) of fatty acids (0.16 g of fatty acids per g of dry weight). These optimized media contained 0.5 g L(-1) of urea or ammonium sulfate and 20 g L(-1) of glycerol, and were devoid of yeast extract. Moreover, Y. lipolytica was engineered by inserting the FatB2 gene, coding for the CpFatB2 thioesterase from Cuphea palustris, in order to modify the fatty acid composition towards the accumulation of medium-chain fatty acids. Contrary to the expected, the expression of the heterologous gene increased the production of oleic acid, and concomitantly decreased the level of saturated fatty acids. PMID:26518537

  10. A Highly Efficient Recombinant Laccase from the Yeast Yarrowia lipolytica and Its Application in the Hydrolysis of Biomass

    PubMed Central

    Kalyani, Dayanand; Tiwari, Manish Kumar; Li, Jinglin; Kim, Sun Chang; Kalia, Vipin C.; Kang, Yun Chan; Lee, Jung-Kul

    2015-01-01

    A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s-1 μM-1) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s-1 μM-1) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification. PMID:25781945

  11. Microbial lipid produced by Yarrowia lipolytica QU21 using industrial waste: a potential feedstock for biodiesel production.

    PubMed

    Poli, Jandora Severo; da Silva, Mirra Angelina Neres; Siqueira, Ezequias P; Pasa, Vânya M D; Rosa, Carlos Augusto; Valente, Patricia

    2014-06-01

    This study aimed to evaluate the effect of medium composition and culture conditions on lipid content, fatty acid profile and biomass production by the yeast Yarrowia lipolytica QU21. Lipid production by the yeast growing on glycerol/(NH4)2SO4 (10%/0.1%) reached 1.48g/L (30.1% according to total cell dry weight). When glycerol was replaced by crude glycerol (industrial waste), the lipid yield was 1.27g/L, with no significant difference. Some particular fatty acids were found when crude glycerol was combined with fresh yeast extract (FYE, brewery waste), as linolenic acid (C18:3n3), eicosadienoic acid (C20:2), eicosatrienoic acid (C20:3n3) and eicosapentaenoic acid (C20:5n3). In addition, the FYE promoted an increase of more than 300% on polyunsaturated fatty acid content (PUFA), which is an undesirable feature for biodiesel production. The fatty acid composition of the oil produced by Y. lipolytica QU21 growing on crude glycerol/(NH4)2SO4 presented a potential use as biodiesel feedstock, with low PUFA content. PMID:24727354

  12. Alcohol dehydrogenases and an alcohol oxidase involved in the assimilation of exogenous fatty alcohols in Yarrowia lipolytica.

    PubMed

    Iwama, Ryo; Kobayashi, Satoshi; Ohta, Akinori; Horiuchi, Hiroyuki; Fukuda, Ryouichi

    2015-05-01

    The yeast Yarrowia lipolytica can assimilate hydrophobic substrates, including n-alkanes and fatty alcohols. Here, eight alcohol dehydrogenase genes, ADH1-ADH7 and FADH, and a fatty alcohol oxidase gene, FAO1, were analyzed to determine their roles in the metabolism of hydrophobic substrates. A mutant deleted for all of these genes (ALCY02 strain) showed severely defective growth on fatty alcohols, and enhanced sensitivity to fatty alcohols in glucose-containing media. The ALCY02 strain grew normally on n-tetradecane or n-hexadecane, but exhibited slightly defective growth on n-decane or n-dodecane. It accumulated more 1-dodecanol and less dodecanoic acid than the wild-type strain when n-dodecane was fed. Expression of ADH1, ADH3 or FAO1, but not that of other ADH genes or FADH, in the ALCY02 strain restored its growth on fatty alcohols. In addition, a triple deletion mutant of ADH1, ADH3 and FAO1 showed similarly defective growth on fatty alcohols and on n-dodecane to the ALCY02 strain. Microscopic observation suggests that Adh1p and Adh3p are localized in the cytosol and Fao1p is in the peroxisome. These results suggest that Adh1p, Adh3p and Fao1p are responsible for the oxidation of exogenous fatty alcohols but play less prominent roles in the oxidation of fatty alcohols derived from n-alkanes. PMID:25805841

  13. Synthetic RNA Polymerase III Promoters Facilitate High-Efficiency CRISPR-Cas9-Mediated Genome Editing in Yarrowia lipolytica.

    PubMed

    Schwartz, Cory M; Hussain, Murtaza Shabbir; Blenner, Mark; Wheeldon, Ian

    2016-04-15

    The oleaginous yeast Yarrowia lipolytica is a valuable microbial host for chemical production because it has a high capacity to synthesize, modify, and store intracellular lipids; however, rapid strain development has been hampered by the limited availability of genome engineering tools. We address this limitation by adapting the CRISPR-Cas9 system from Streptococcus pyogenes for markerless gene disruption and integration in Y. lipolytica. Single gene disruption efficiencies of 92% and higher were achieved when single guide RNAs (sgRNA) were transcribed with synthetic hybrid promoters that combine native RNA polymerase III (Pol III) promoters with tRNA. The Pol III-tRNA hybrid promoters exploit endogenous tRNA processing to produce mature sgRNA for Cas9 targeting. The highest efficiencies were achieved with a SCR1'-tRNA(Gly) promoter and Y. lipolytica codon-optimized Cas9 expressed from a UAS1B8-TEF promoter. Cotransformation of the Cas9 and sgRNA expressing plasmid with a homologous recombination donor plasmid resulted in markerless homologous recombination efficiency of over 64%. Homologous recombination was observed in 100% of transformants when nonhomologous end joining was disrupted. The end result of these studies was the development of pCRISPRyl, a modular tool for markerless gene disruption and integration in Y. lipolytica. PMID:26714206

  14. Comprehensive metabolomic, lipidomic and microscopic profiling of Yarrowia lipolytica during lipid accumulation identifies targets for increased lipogenesis

    DOE PAGESBeta

    Pomraning, Kyle R.; Wei, Siwei; Karagiosis, Sue A.; Kim, Young-Mo; Dohnalkova, Alice; Arey, Bruce W.; Bredeweg, Erin L.; Orr, Galya; Metz, Thomas O.; Baker, Scott E.

    2015-04-23

    Yarrowia lipolytica is an oleaginous ascomycete yeast that accumulates large amounts of lipids and has potential as a biofuel producing organism. Despite a growing scientific literature focused on lipid production by Y. lipolytica, there remain significant knowledge gaps regarding the key biological processes involved. We applied a combination of metabolomic and lipidomic profiling approaches as well as microscopic techniques to identify and characterize the key pathways involved in de novo lipid accumulation from glucose in batch cultured, wild-type Y. lipolytica. We found that lipids accumulated rapidly and peaked at 48 hours during the five day experiment, concurrent with a shiftmore » in amino acid metabolism. We also report that Y. lipolytica secretes disaccharides early in batch culture and reabsorbs them when extracellular glucose is depleted. Exhaustion of extracellular sugars coincided with thickening of the cell wall, suggesting that genes involved in cell wall biogenesis may be a useful target for improving the efficiency of lipid producing yeast strains.« less

  15. van der Waals torque

    NASA Astrophysics Data System (ADS)

    Esquivel-Sirvent, Raul; Schatz, George

    2014-03-01

    The theory of generalized van der Waals forces by Lifshtz when applied to optically anisotropic media predicts the existence of a torque. In this work we present a theoretical calculation of the van der Waals torque for two systems. First we consider two isotropic parallel plates where the anisotropy is induced using an external magnetic field. The anisotropy will in turn induce a torque. As a case study we consider III-IV semiconductors such as InSb that can support magneto plasmons. The calculations of the torque are done in the Voigt configuration, that occurs when the magnetic field is parallel to the surface of the slabs. The change in the dielectric function as the magnetic field increases has the effect of decreasing the van der Waals force and increasing the torque. Thus, the external magnetic field is used to tune both the force and torque. The second example we present is the use of the torque in the non retarded regime to align arrays of nano particle slabs. The torque is calculated within Barash and Ginzburg formalism in the nonretarded limit, and is quantified by the introduction of a Hamaker torque constant. Calculations are conducted between anisotropic slabs of materials including BaTiO3 and arrays of Ag nano particles. Depending on the shape and arrangement of the Ag nano particles the effective dielectric function of the array can be tuned as to make it more or less anisotropic. We show how this torque can be used in self assembly of arrays of nano particles. ref. R. Esquivel-Sirvent, G. C. Schatz, Phys. Chem C, 117, 5492 (2013). partial support from DGAPA-UNAM.

  16. Zeit im Wandel der Zeit.

    NASA Astrophysics Data System (ADS)

    Aichelburg, P. C.

    Contents: Einleitung(P. C. Aichelburg). 1. Über Zeit, Bewegung und Veränderung (Aristoteles). 2. Ewigkeit und Zeit (Plotin). 3. Was ist die Zeit? (Augustinus). 4. Von der Zeit (Immanuel Kant). 5. Newtons Ansichten über Zeit, Raum und Bewegung (Ernst Mach). 6. Über die mechanische Erklärung irreversibler Vorgänge (Ludwig Boltzmann). 7. Das Maß der Zeit (Henri Poincaré). 8. Dauer und Intuition (Henri Bergson). 9. Die Geschichte des Unendlichkeitsproblems (Bertrand Russell). 10. Raum und Zeit (Hermann Minkowski). 11. Der Unterschied von Zeit und Raum (Hans Reichenbach). 12. Newtonscher und Bergsonscher Zeitbegriff (Norbert Wiener). 13. Die Bildung des Zeitbegriffs beim Kinde (JeanPiaget).14. Eine Bemerkung über die Beziehungen zwischen Relativitätstheorie und der idealistischen Philosophie (Kurt Gödel). 15. Der zweite Hauptsatz und der Unterschied von Vergangenheit und Zukunft (Carl Friedrich v. Weizsäcker). 16. Zeit als physikalischer Begriff (Friedrich Hund). 17. Zeitmessung und Zeitbegriff in der Astronomie (Otto Heckmann). 18. Kann die Zeit rückwärts gehen? (Martin Gardner). 19. Zeit und Zeiten (Ilya Prigogine, Isabelle Stengers). 20. Zeit als dynamische Größe in der Relativitätstheorie (P. C. Aichelburg).

  17. Das Lob der Sternkunst. Astronomie in der deutschen Aufklärung.

    NASA Astrophysics Data System (ADS)

    Baasner, R.

    Contents: 1. Einleitung. 2. Die Astronomie im Rahmen der Aufklärungs-Physik. 3. Das Lob der Sternkunst. 4. Ein Blick auf die Sternwarten. 5. Allgemeine Darstellungen der Sternkunde. 6. Schleppende Rezeption: Das kopernikanische Weltbild. 7. Himmelsphysik: Die Debatte um die causa gravitatis. 8. Theorie der Himmelskörper. 9. Die Erde als Gegenstand der Astronomie. 10. Die Sonne. 11. Der Mond. 12. Die Planeten. 13. Die Kometen. 14. Die Fixsterne. 15. Die Entstehung der Welt. 16. Beiträge der Astrotheologie. 17. Der Kampf gegen die Astrologen.

  18. Biochips und ihr Einsatz in der Lebensmittelanalytik

    NASA Astrophysics Data System (ADS)

    Huber, Ingrid; Zeltz, Patric

    Mit der Verbreitung des Begriffes "Biochip“ in den biotechnologischen Medien wurde Ende der 1990er-Jahre zunächst der Eindruck erweckt, dass die Computerelektronik in die molekularbiologischen Anwendungen eingestiegen ist [18]. In nur wenigen Jahren hat sich die Biochiptechnologie zu einem Verfahren entwickelt, das aus der molekularbiologischen Grundlagenforschung nicht mehr wegzudenken ist und über eine Vielzahl von Einsatzbereichen verfügt. Die Biochiptechnologie ermöglicht die Miniaturisierung von DNA-, RNA- bzw. Proteinanalytik in hochparallelen Formaten. Dieser hohe Parallelisierungsgrad ist einer der wesentlichen Vorteile dieser Technik gegenüber klassischen molekularbiologischen Methoden. Sie wird heutzutage vor allem in der Genomforschung eingesetzt, für Genexpressionsstudien, zum Screening von single nucleotide polymorphisms (SNPs), in der pharmakogenetischen Forschung sowie in der Erforschung von Erbkrankheiten und in der Krebsforschung [1, 7, 19]. Neben vielen weiteren Bereichen finden Biochips auch spezielle Anwendungen in der Lebensmittelanalytik.

  19. Dialektischer Materialismus in der Quantentheorie

    NASA Astrophysics Data System (ADS)

    Fuchs, Klaus

    Der absolute Determinismus der klassischen Mechanik bietet keine Ansatzpunkte für eine befriedigende Naturphilosophie. Mit der Quantenmechanik werden nicht lediglich die Unzulänglichkeiten einzelner klassischer Begriffe, sondern die des gesamten klassischen Begriffssystems beseitigt.Translated AbstractDialectical Materialism in Quantum TheoryThe absolute determinism of classical mechanics does not provide any base for a satisfactory philosophy of nature. In quantum mechanics the shortcomings of not only some single classical concepts but of the classical description as a whole are removed.

  20. High performance microbiological transformation of L-tyrosine to L-dopa by Yarrowia lipolytica NRRL-143

    PubMed Central

    Ali, Sikander; Shultz, Jeffry L; Ikram-ul-Haq

    2007-01-01

    Background The 3,4-dihydroxy phenyl L-alanine (L-dopa) is a drug of choice for Parkinson's disease, controlling changes in energy metabolism enzymes of the myocardium following neurogenic injury. Aspergillus oryzae is commonly used for L-dopa production; however, potential improvements in ease of handling, growth rate and environmental impact have led to an interest in exploiting alternative yeasts. The two important elements required for L-dopa production are intracellular tyrosinases (thus pre-grown yeast cells are required for the transformation of L-tyrosine to L-dopa) and L-ascorbate, which acts as a reducing agent. Results Pre-grown cells of Yarrowia lipolytica NRRL-143 were used for the microbiological transformation of L-tyrosine to L-dopa. Different diatomite concentrations (0.5–3.0 mg/ml) were added to the acidic (pH 3.5) reaction mixture. Maximum L-dopa biosynthesis (2.96 mg/ml L-dopa from 2.68 mg/ml L-tyrosine) was obtained when 2.0 mg/ml diatomite was added 15 min after the start of the reaction. After optimizing reaction time (30 min), and yeast cell concentration (2.5 mg/ml), an overall 12.5 fold higher L-dopa production rate was observed when compared to the control. Significant enhancements in Yp/s, Qs and qs over the control were observed. Conclusion Diatomite (2.0 mg/ml) addition 15 min after reaction commencement improved microbiological transformation of L-tyrosine to L-dopa (3.48 mg/ml; p ≤ 0.05) by Y. lipolytica NRRL-143. A 35% higher substrate conversion rate was achieved when compared to the control. PMID:17705832

  1. Role of pyruvate carboxylase in accumulation of intracellular lipid of the oleaginous yeast Yarrowia lipolytica ACA-DC 50109.

    PubMed

    Wang, Guang-Yuan; Zhang, Yan; Chi, Zhe; Liu, Guang-Lei; Wang, Zhi-Peng; Chi, Zhen-Ming

    2015-02-01

    Yarrowia lipolytica ACA-DC 50109 is an oleaginous yeast. In order to know the function of pyruvate carboxylase (PYC) in lipid biosynthesis, the PYC gene cloned from Pichia guilliermondii Pcla22 was overexpressed in the oleaginous yeast. The lipid contents in the wild-type strain ACA-DC 50109 and the transformants P4, P7, and P103 were 30.2 % (w/w) 36.5 % (w/w), 38.2 % (w/w), and 37.9 % (w/w). However, the amount of the secreted citric acids by strains ACA-DC 50109, P4, P77, and P103 were 0.5, 10.1, 11.5, and 9.4 g/L. In order to reduce the amount of the secreted citric acid, the PYC gene and endogenous ACL1 gene encoding ATP citrate lyase (ACL1) were simultaneously overexpressed in the oleaginous yeast. The lipid contents of the transformants PA19, PA56, PA124 were 44.4 % (w/w), 45.3 % (w/w), and 43.7 % (w/w). At the same time, the amount of the secreted citric acid by the transformants PA19, PA56, and PA124 was reduced to 5.4, 6.2, and 6.3 g/L. The PYC and ACL1 activities and their gene transcriptional levels in all the transformants were greatly enhanced compared to those in their wild-type strain ACA-DC 50109. During 10-L fermentation, lipid content in the transformant PA56 was 49.6 % (w/w) and the amount of secreted citric acid was 2.9 g/L. This meant that PYC and ACL1 can play an important role in accumulation of intracellular lipid of the oleaginous yeast Y. lipolytica ACA-DC 50109. PMID:25427679

  2. Biochemical characterization of Yarrowia lipolytica LIP8, a secreted lipase with a cleavable C-terminal region.

    PubMed

    Kamoun, Jannet; Schué, Mathieu; Messaoud, Wala; Baignol, Justine; Point, Vanessa; Mateos-Diaz, Eduardo; Mansuelle, Pascal; Gargouri, Youssef; Parsiegla, Goetz; Cavalier, Jean-François; Carrière, Frédéric; Aloulou, Ahmed

    2015-02-01

    Yarrowia lipolytica is a lipolytic yeast possessing 16 paralog genes coding for lipases. Little information on these lipases has been obtained and only the major secreted lipase, namely YLLIP2, had been biochemically and structurally characterized. Another secreted lipase, YLLIP8, was isolated from Y. lipolytica culture medium and compared with the recombinant enzyme produced in Pichia pastoris. N-terminal sequencing showed that YLLIP8 is produced in its active form after the cleavage of a signal peptide. Mass spectrometry analysis revealed that YLLIP8 recovered from culture medium lacks a C-terminal part of 33 amino acids which are present in the coding sequence. A 3D model of YLLIP8 built from the X-ray structure of the homologous YLLIP2 lipase shows that these truncated amino acids in YLLIP8 belong to an additional C-terminal region predicted to be mainly helical. Western blot analysis shows that YLLIP8 C-tail is rapidly cleaved upon enzyme secretion since both cell-bound and culture supernatant lipases lack this extension. Mature recombinant YLLIP8 displays a true lipase activity on short-, medium- and long-chain triacylglycerols (TAG), with an optimum activity at alkaline pH on medium chain TAG. It has no apparent regioselectivity in TAG hydrolysis, thus generating glycerol and FFAs as final lipolysis products. YLLIP8 properties are distinct from those of the 1,3-regioselective YLLIP2, acting optimally at acidic pH. These lipases are tailored for complementary roles in fatty acid uptake by Y. lipolytica. PMID:25449652

  3. Four distinct secretory pathways serve protein secretion, cell surface growth, and peroxisome biogenesis in the yeast Yarrowia lipolytica.

    PubMed Central

    Titorenko, V I; Ogrydziak, D M; Rachubinski, R A

    1997-01-01

    We have identified and characterized mutants of the yeast Yarrowia lipolytica that are deficient in protein secretion, in the ability to undergo dimorphic transition from the yeast to the mycelial form, and in peroxisome biogenesis. Mutations in the SEC238, SRP54, PEX1, PEX2, PEX6, and PEX9 genes affect protein secretion, prevent the exit of the precursor form of alkaline extracellular protease from the endoplasmic reticulum, and compromise peroxisome biogenesis. The mutants sec238A, srp54KO, pex2KO, pex6KO, and pex9KO are also deficient in the dimorphic transition from the yeast to the mycelial form and are affected in the export of only plasma membrane and cell wall-associated proteins specific for the mycelial form. Mutations in the SEC238, SRP54, PEX1, and PEX6 genes prevent or significantly delay the exit of two peroxisomal membrane proteins, Pex2p and Pex16p, from the endoplasmic reticulum en route to the peroxisomal membrane. Mutations in the PEX5, PEX16, and PEX17 genes, which have previously been shown to be essential for peroxisome biogenesis, affect the export of plasma membrane and cell wall-associated proteins specific for the mycelial form but do not impair exit from the endoplasmic reticulum of either Pex2p and Pex16p or of proteins destined for secretion. Biochemical analyses of these mutants provide evidence for the existence of four distinct secretory pathways that serve to deliver proteins for secretion, plasma membrane and cell wall synthesis during yeast and mycelial modes of growth, and peroxisome biogenesis. At least two of these secretory pathways, which are involved in the export of proteins to the external medium and in the delivery of proteins for assembly of the peroxisomal membrane, diverge at the level of the endoplasmic reticulum. PMID:9271399

  4. The oxidative pentose phosphate pathway is the primary source of NADPH for lipid overproduction from glucose in Yarrowia lipolytica.

    PubMed

    Wasylenko, Thomas M; Ahn, Woo Suk; Stephanopoulos, Gregory

    2015-07-01

    Oleaginous microbes represent an attractive means of converting a diverse range of feedstocks into oils that can be transesterified to biodiesel. However, the mechanism of lipid overproduction in these organisms is incompletely understood, hindering the development of strategies for engineering superior biocatalysts for "single-cell oil" production. In particular, it is unclear which pathways are used to generate the large quantities of NADPH required for overproduction of the highly reduced fatty acid species. While early studies implicated malic enzyme as having a key role in production of lipogenic NADPH in oleaginous fungi, several recent reports have cast doubts as to whether malic enzyme may contribute to production of lipogenic NADPH in the model oleaginous yeast Yarrowia lipolytica. To address this problem we have used (13)C-Metabolic Flux Analysis to estimate the metabolic flux distributions during lipid accumulation in two Y. lipolytica strains; a control strain and a previously published engineered strain capable of producing lipids at roughly twice the yield. We observe a dramatic rearrangement of the metabolic flux distribution in the engineered strain which supports lipid overproduction. The NADPH-producing flux through the oxidative Pentose Phosphate Pathway is approximately doubled in the engineered strain in response to the roughly two-fold increase in fatty acid biosynthesis, while the flux through malic enzyme does not differ significantly between the two strains. Moreover, the estimated rate of NADPH production in the oxidative Pentose Phosphate Pathway is in good agreement with the estimated rate of NADPH consumption in fatty acid biosynthesis in both strains. These results suggest the oxidative Pentose Phosphate Pathway is the primary source of lipogenic NADPH in Y. lipolytica. PMID:25747307

  5. Three alcohol dehydrogenase genes and one acetyl-CoA synthetase gene are responsible for ethanol utilization in Yarrowia lipolytica.

    PubMed

    Gatter, Michael; Ottlik, Stephanie; Kövesi, Zsolt; Bauer, Benjamin; Matthäus, Falk; Barth, Gerold

    2016-10-01

    The non-conventional yeast Yarrowia lipolytica is able to utilize a wide range of different substrates like glucose, glycerol, ethanol, acetate, proteins and various hydrophobic molecules. Although most metabolic pathways for the utilization of these substrates have been clarified by now, it was not clear whether ethanol is oxidized by alcohol dehydrogenases or by an alternative oxidation system inside the cell. In order to detect the genes that are required for ethanol utilization in Y. lipolytica, eight alcohol dehydrogenase (ADH) genes and one alcohol oxidase gene (FAO1) have been identified and respective deletion strains were tested for their ability to metabolize ethanol. As a result of this, we found that the availability of ADH1, ADH2 or ADH3 is required for ethanol utilization in Y. lipolytica. A strain with deletions in all three genes is lacking the ability to utilize ethanol as sole carbon source. Although Adh2p showed by far the highest enzyme activity in an in vitro assay, the availability of any of the three genes was sufficient to enable a decent growth. In addition to ADH1, ADH2 and ADH3, an acetyl-CoA synthetase encoding gene (ACS1) was found to be essential for ethanol utilization. As Y. lipolytica is a non-fermenting yeast, it is neither able to grow under anaerobic conditions nor to produce ethanol. To investigate whether Y. lipolytica may produce ethanol, the key genes of alcoholic fermentation in S. cerevisiae, ScADH1 and ScPDC1, were overexpressed in an ADH and an ACS1 deletion strain. However, instead of producing ethanol, the respective strains regained the ability to use ethanol as single carbon source and were still not able to grow under anaerobic conditions. PMID:27486067

  6. Der evolutionäre Naturalismus in der Ethik

    NASA Astrophysics Data System (ADS)

    Kaiser, Marie I.

    Charles Darwin hat eindrucksvoll gezeigt, dass der Mensch ebenso wie alle anderen Lebewesen ein Produkt der biologischen Evolution ist. Die sich an Darwin anschließende Forschung hat außerdem plausibel gemacht, dass sich nicht nur viele der körperlichen Merkmale des Menschen, sondern auch (zumindest einige) seiner Verhaltensdispositionen in adaptiven Selektionsprozessen herausgebildet haben. Die Vorstellung, dass auch die menschliche Moralität evolutionär bedingt ist, scheint daher auf den ersten Blick ganz überzeugend. Schließlich hat die Evolutionstheorie in den vergangenen Jahrzehnten in vielen Bereichen (auch außerhalb der Biologie) ihre weitreichende Bedeutung unter Beweis gestellt. Warum sollte, so könnte man beispielsweise fragen, gerade die Fähigkeit des Menschen, moralische Normen aufzustellen und gemäß ihnen zu handeln, nicht evolutionär erklärt werden können? Und warum sollte eine solche evolutionäre Erklärung der menschlichen Moralität irrelevant für die Rechtfertigung moralischer Normen sein? Warum sollte die Ethik eine Bastion der Philosophen bleiben, für die evolutionsbiologische Forschungsergebnisse über den Menschen und seine nächsten Verwandten keinerlei Relevanz besitzen?

  7. An ortholog of farA of Aspergillus nidulans is implicated in the transcriptional activation of genes involved in fatty acid utilization in the yeast Yarrowia lipolytica

    SciTech Connect

    Poopanitpan, Napapol; Kobayashi, Satoshi; Fukuda, Ryouichi; Horiuchi, Hiroyuki; Ohta, Akinori

    2010-11-26

    Research highlights: {yields} POR1 is a Yarrowia lipolytica ortholog of farA involved in fatty acid response in A. nidulans. {yields} Deletion of POR1 caused growth defects on fatty acids. {yields} {Delta}por1 strain exhibited defects in the induction of genes involved in fatty acid utilization. -- Abstract: The yeast Yarrowia lipolytica effectively utilizes hydrophobic substrates such as fatty acids and n-alkanes. To identify a gene(s) regulating fatty acid utilization in Y. lipolytica, we first studied homologous genes to OAF1 and PIP2 of Saccharomyces cerevisiae, but their disruption did not change growth on oleic acid at all. We next characterized a Y. lipolytica gene, POR1 (primary oleate regulator 1), an ortholog of farA encoding a transcriptional activator that regulates fatty acid utilization in Aspergillus nidulans. The deletion mutant of POR1 was defective in the growth on various fatty acids, but not on glucose, glycerol, or n-hexadecane. It exhibited slight defect on n-decane. The transcriptional induction of genes involved in {beta}-oxidation and peroxisome proliferation by oleate was distinctly diminished in the {Delta}por1 strains. These data suggest that POR1 encodes a transcriptional activator widely regulating fatty acid metabolism in Y. lipolytica.

  8. Design of an efficient medium for heterologous protein production in Yarrowia lipolytica: case of human interferon alpha 2b

    PubMed Central

    2011-01-01

    Background The non conventional yeast Yarrowia lipolytica has aroused a strong industrial interest for heterologous protein production. However most of the studies describing recombinant protein production by this yeast rely on the use of complex media, such media are not convenient for large scale production particularly for products intended for pharmaceutical applications. In addition medium composition can also affect the production yield. Hence it is necessary to design an efficient medium for therapeutic protein expression by this host. Results Five different media, including four minimal media and a complex medium, were assessed in shake flasks for the production of human interferon alpha 2b (hIFN α2b) by Y. lipolytica under the control of POX2 promoter inducible with oleic acid. The chemically defined medium SM4 formulated by Invitrogen for Pichia pastoris growth was the most suitable. Using statistical experimental design this medium was further optimized. The selected minimal medium consisting in SM4 supplemented with 10 mg/l FeCl3, 1 g/l glutamate, 5 ml/l PTM1 (Pichia Trace Metals) solution and a vitamin solution composed of myo-inositol, thiamin and biotin was called GNY medium. Compared to shake flask, bioreactor culture in GNY medium resulted in 416-fold increase of hIFN α2b production and 2-fold increase of the biological activity. Furthermore, SM4 enrichment with 5 ml/l PTM1 solution contributed to protect hIFN α2b against the degradation by the 28 kDa protease identified by zymography gel in culture supernatant. The screening of the inhibitory effect of the trace elements present in PTM1 solution on the activity of this protease was achieved using a Box-Behnken design. Statistical data analysis showed that FeCl3 and MnSO4 had the most inhibitory effect. Conclusion We have designed an efficient medium for large scale production of heterologous proteins by Y. lipolytica. The optimized medium GNY is suitable for the production of hIFN α2b with the

  9. Der Physik-Nobelpreis vor 100 Jahren Eine neue Eigenschaft der Materie

    NASA Astrophysics Data System (ADS)

    Jacobi, Manfred

    2003-11-01

    Im Jahre 1903 erhielten Henri Becquerel sowie Marie und Pierre Curie den Physik-Nobelpreis für die Entdeckung der Radioaktivität. Schon damals stellte Pierre Curie die Frage nach Nutzen und Gefahr dieser neuen Eigenschaft von Materie. Er war der Überzeugung, dass der Fortschritt letztlich mehr zum Nutzen als zum Schaden der Menschen beitrage.

  10. Physik gestern und heute Der Spion, der die Wärme untersuchte

    NASA Astrophysics Data System (ADS)

    Heering, Peter

    2003-11-01

    Der in Massachusetts geborene Benjamin Thompson, der ab 1792 auch den Titel eines Grafen von Rumford führte, ist eine der schillerndsten Figuren der Physikgeschichte. Berühmt wurde er insbesondere wegen seiner Experimente zur Wärmetheorie.http://www.famousamericans.net/benjaminthompsonrumford

  11. Coronellis Cosmos in der Melker Stiftsbibliothek.

    NASA Astrophysics Data System (ADS)

    Glaßner, Gottfried; Pärr, Nora

    2009-06-01

    Die Melker Stiftsbibliothek besitzt ein Globenpaar des berühmten venezianischen Globenbauers Vincenzo Coronelli (1650-1718), einen Erdglobus von 1688 und einen Himmelsglobus von 1693. Wie und wann die beiden Globen nach Melk gekommen sind, ist nicht bekannt. Dass sie zur ursprünglichen Ausstattung der 1735 fertig gestellten Barockbibliothek gehörten, wird aber aus der zentralen Stellung deutlich, die dem Globus (Erdglobus und Armillarphäre) in dem von Paul Troger 1732 gemalten Deckenfresko zukommt. Mehrfach begegnet das Motiv des Globus als Attribut der Weisheit bzw. Philosophie, der Geographie bzw. Geometrie und der Astronomie in den beiden Hauptsälen wie auch in der Deckenmalerei von Johann Bergl in der Oberen Bibliothek (1768) und in der Kuppel des Gartenpavillons (1764).

  12. Historisches Rätsel Der rastlose Amerikaner

    NASA Astrophysics Data System (ADS)

    Loos, Andreas

    2004-09-01

    In der Schule, die er mit acht Jahren zum ersten Mal von innen sah, hielt man ihn nicht für allzu helle - schließlich hinkte der Kleine oft hinter der Klasse her. Und es hat etwas Tragisches, wenn der wohl berühmteste Erfinder aller Zeiten und Halter von 2000 Patenten im Alter über seinen Vater sagt: Er dachte, ich sei dumm. Und ich meinte schon fast selbst, ich sei ein Dummkopf.

  13. Designing of a "cheap to run" fermentation platform for an enhanced production of single cell oil from Yarrowia lipolytica DSM3286 as a potential feedstock for biodiesel.

    PubMed

    Nambou, Komi; Zhao, Chen; Wei, Liujing; Chen, Jun; Imanaka, Tadayuki; Hua, Qiang

    2014-12-01

    In this study, the culture medium components screening and filtering were undertaken in order to set up efficient and cost effective minimal culture media for lipid production from Yarrowia lipolytica DSM3286. The basal minimal culture medium (S2) designed yielded lipid content up to 35% of the microbial dry cell weight. A set of fermentation strategies based on this minimal medium was developed and the lipid content was raised to 51%. The scale-up under different fermentation conditions based on S2 medium led to a maximum lipid content of 65%. The produced microbial oils displayed interesting properties to be used as a feedstock for high quality biodiesel production. The minimal media and operable cultivation strategies devised in this study, in association with the works done so far by other authors, could enable fast, massive, viable and more economical production of single cell oils and smooth biodiesel manufacture. PMID:25310869

  14. Microwave, ultrasound, thermal treatments, and bead milling as intensification techniques for extraction of lipids from oleaginous Yarrowia lipolytica yeast for a biojetfuel application.

    PubMed

    Meullemiestre, Alice; Breil, Cassandra; Abert-Vian, Maryline; Chemat, Farid

    2016-07-01

    In the present work, two different ways of lipids extraction from Yarrowia lipolytica yeast were investigated in order to maximize the extraction yield. Firstly, various modern techniques of extraction including ultrasound, microwave, and bead milling were tested to intensify the efficiency of lipid recovery. Secondly, several pretreatments such as freezing/defrosting, cold drying, bead milling, and microwave prior two washing of mixture solvent of chloroform:methanol (1:2, v/v) were study to evaluate the impact on lipid recovery. All these treatments were compared to conventional maceration, in terms of lipids extraction yield and lipid composition analysis. The main result of this study is the large difference of lipid recovery among treatments and the alteration of lipids profile after microwave and ultrasound techniques. PMID:27017129

  15. [Study of the Accumulation of Rec A from Bacillus subtilis in the Mitochondria of a Recombinant Strain of the Yeast Yarrowia lipolytica].

    PubMed

    Isakova, E P; Deryabina, Y I; Leonovich, O A; Zylkova, M V; Biriukova, Iu K

    2016-01-01

    No eukaryotic species has a system for homologous DNA recombination of the mitochondrial genome. We report on an integrative genetic systembased on the pQ-SRUS construct that allows the expression of the RecA recombinase from Bacillus subtilis and its transportation to mitochondria of Yarrowia lipolytica. The targeting of recombinant RecA to mitochondria is provided by leader sequences (5'-UTR and 3-UTR) derived from the SOD2 gene mRNA, which exhibit affinity to the outer mitochondrial membrane and provides cotranslational import of RecA to the inner space of mitochondria. The accumulation of RecA in mitochondria of the Y. lipolytica recombinant strain bearing the pQ-SRUS construct has been shown by immunoblotting of purified mitochondrial preparations. PMID:27266246

  16. Integrated Approach To Producing High-Purity Trehalose from Maltose by the Yeast Yarrowia lipolytica Displaying Trehalose Synthase (TreS) on the Cell Surface.

    PubMed

    Li, Ning; Wang, Hengwei; Li, Lijuan; Cheng, Huiling; Liu, Dawen; Cheng, Hairong; Deng, Zixin

    2016-08-10

    An alternative strategy that integrated enzyme production, trehalose biotransformation, and bioremoval in one bioreactor was developed in this study, thus simplifying the traditional procedures used for trehalose production. The trehalose synthase gene from a thermophilic archaea, Picrophilus torridus, was first fused to the YlPir1 anchor gene and then inserted into the genome of Yarrowia lipolytica, thus yielding an engineered yeast strain. The trehalose yield reached 73% under optimal conditions. The thermal and pH stabilities of the displayed enzyme were improved compared to those of its free form purified from recombinant Escherichia coli. After biotransformation, the glucose byproduct and residual maltose were directly fermented to ethanol by a Saccharomyces cerevisiae strain. Ethanol can be separated by distillation, and high-purity trehalose can easily be obtained from the fermentation broth. The results show that this one-pot procedure is an efficient approach to the economical production of trehalose from maltose. PMID:27472444

  17. [The 'van der Linden' retainer].

    PubMed

    van der Linden, F P

    2003-11-01

    Normally, only the anterior teeth have to be retained after an orthodontic treatment. For that purpose, a lingually bonded wire is preferred in the mandible and a removable plate in the maxilla. The design of the Van der Linden-retainer is based on theoretical considerations and secures rigid fixation of the six anterior teeth with solid anchorage in that region. A retention plate should not be used to move anterior teeth. However, with instant corrections small improvements can be realized. The fabrication and clinical use of the Van der Linden-retainer is explained and illustrated. PMID:14669487

  18. Development of a cultivation process for the enhancement of human interferon alpha 2b production in the oleaginous yeast, Yarrowia lipolytica

    PubMed Central

    2011-01-01

    Background As an oleaginous yeast, Yarrowia lipolytica is able to assimilate hydrophobic substrates. This led to the isolation of several promoters of key enzymes of this catabolic pathway. Less is known about the behavior of Y. lipolytica in large bioreactors using these substrates. There is therefore a lack of established know-how concerning high cell density culture protocols of this yeast. Consequently, the establishment of suitable induction conditions is required, to maximize recombinant protein production under the control of these promoters. Results Human interferon α2b (huIFN α2b) production in Yarrowia lipolytica was used as a model for the enhancement of recombinant protein production under the control of the oleic acid (OA)-inducible promoter POX2. Cell viability and heterologous protein production were enhanced by exponential glucose feeding, to generate biomass before OA induction. The optimal biomass level before induction was determined (73 g L-1), and glucose was added with oleic acid during the induction phase. Several oleic acid feeding strategies were assessed. Continuous feeding with OA at a ratio of 0.02 g OA per g dry cell weight increased huIFNα2b production by a factor of 1.88 (425 mg L-1) and decreased the induction time (by a factor of 2.6, 21 h). huIFN α2b degradation by an aspartic protease secreted by Y. lipolytica was prevented by adding pepstatin (10 μM), leading to produce a 19-fold more active huIFN α2b (26.2 × 107 IU mg-1). Conclusion Y. lipolytica, a generally regarded as safe (GRAS) microorganism is one of the most promising non conventional yeasts for the production of biologically active therapeutic proteins under the control of hydrophobic substrate-inducible promoter. PMID:22047602

  19. Der Begriff der Verständlichkeit in der modernen Physik (1948).

    PubMed

    Feyerabend, Paul K

    2016-06-01

    This is a critical transcription of Paul Feyerabend's earliest extant essay "Der Begriff der Verständlichkeit in der modernen Physik" (1948) recovered from the European Forum Alpbach archives. In it, Feyerabend defends positivism as a progressive framework for scientific research in certain stages of scientific development. He argues that in physics visualizability (Anschaulichkeit) and intelligibility (Verständlichkeit) are time-conditioned concepts: what is deemed visualizable in the development of physical theories is relative to a specific historical context and changes over time. He concludes that from time to time the abandonment of visualizability is crucial for progress in physics, as it is conducive to major theory change, illustrating the point on the basis of advances in atomic theory. PMID:27269264

  20. Ancillary Services Provided from DER

    SciTech Connect

    Campbell, J.B.

    2005-12-21

    Distributed energy resources (DER) are quickly making their way to industry primarily as backup generation. They are effective at starting and then producing full-load power within a few seconds. The distribution system is aging and transmission system development has not kept up with the growth in load and generation. The nation's transmission system is stressed with heavy power flows over long distances, and many areas are experiencing problems in providing the power quality needed to satisfy customers. Thus, a new market for DER is beginning to emerge. DER can alleviate the burden on the distribution system by providing ancillary services while providing a cost adjustment for the DER owner. This report describes 10 types of ancillary services that distributed generation (DG) can provide to the distribution system. Of these 10 services the feasibility, control strategy, effectiveness, and cost benefits are all analyzed as in the context of a future utility-power market. In this market, services will be provided at a local level that will benefit the customer, the distribution utility, and the transmission company.

  1. Ressourceneinsatzplanung In der Energie- und Verfahrenstechnik

    NASA Astrophysics Data System (ADS)

    Engell, Sebastian; Handschin, Edmund; Rehtanz, Christian; Schultz, Rüdiger

    Der ökologisch nachhaltige, wirtschaftlich effiziente und politisch faire Umgang mit den Ressourcen der Erde ist eine der größten Herausforderungen an die Menschheit im 21. Jahrhundert und impliziert Paradigmenwechsel in allen Bereichen des gesellschaftlichen Lebens. Energie- und Verfahrenstechnik sind traditionell Bereiche, in denen der optimale Umgang mit kostbaren und beschränkt verfügbaren Ressourcen von erstrangiger Bedeutung ist. Zur Lösung der damit verbundenen komplexen Entscheidungsprobleme sind mathematische Methoden schon seit längerem im Einsatz. Hierbei können aber noch längst nicht alle bislang aufgetretenen Probleme fundiert angegangen werden. Der wachsende wirtschaftliche und ökologische Druck auf die Effizienz der Produktions-und Versorgungssysteme sowie stärkere Schwankungen von Angebot und Nachfrage führen zu neuen Herausforderungen, für die neue mathematische Methoden und Algorithmen entwickelt werden müssen.

  2. Das Neutron, der Kosmos und die Kräfte: Neutronen in der Teilchenphysik

    NASA Astrophysics Data System (ADS)

    Soldner, Torsten

    2003-05-01

    Das Neutron besitzt eine einzigartige Kombination von Eigenschaften. Sie ermöglicht die Untersuchung aller vier elementaren Kräfte. Dabei wurden beeindruckende Resultate erzielt, wie die Präzisionsmessungen der elektrischen Ladung des Neutrons oder der Feinstrukturkonstante zeigen. Die genaue Bestimmung der schwachen Wechselwirkungsstärke der Nukleonen liefert der Astrophysik wichtige Daten. Die Messung eines von Null verschiedenen elektrischen Dipolmoments des Neutrons könnte einen entscheidenden Hinweis über die Physik jenseits des Standardmodells der Teilchenphysik geben. Doch auch zur Aufklärung von Unsicherheiten innerhalb des Standardmodells selbst tragen Neutronen bei.

  3. Verbesserung der Prozessbedingungen beim Einlippentiefbohren durch unterschiedliche Formen der Vorschubmodulation

    NASA Astrophysics Data System (ADS)

    Eisseler, Rocco

    Das Tiefbohren mit Einlippenwerkzeugen ist bei vielen Anwendungen das Verfahren der ersten Wahl, vor allem wenn kleine und kleinste Bohrungsdurchmesser, ein sehr großes Verhältnis zwischen Bohrtiefe und Durchmesser (l/D) und eine hohe Bearbeitungsqualität gefordert werden. Der erreichbare Durchmesserbereich des Verfahrens liegt zwischen D = 0,5 mm und 40 mm, wobei die möglichen Bohrtiefen von l = 3 x D bis 250 x D reichen. Diese Werte beziehen sich auf das Vollbohren, beim Aufbohren können noch größere Durchmesser erreicht werden [1]. Die erreichbare Oberflächenqualität ist so hoch, dass in den meisten Fällen auf eine nachfolgende Feinbearbeitung verzichtet werden kann.

  4. Aufgaben und Genauigkeit der klinischen Dosimetrie

    NASA Astrophysics Data System (ADS)

    Krieger, Hanno

    In diesem Kapitel werden die Aufgaben der klinischen Dosimetrie für die verschiedenen radiologischen Disziplinen zusammengestellt. Die wichtigste Aufgabe ist die Messung der im bestrahlten Medium entstandenen Energiedosis für die verschiedenen Strahlungsquellen. Die am weitesten verbreitete dazu verwendete Methode ist die Dosismessung mit gasgefüllten Ionisationskammern. Im zweiten Teil des Kapitels werden die Genauigkeitsanforderungen der klinischen Dosimetrie diskutiert.

  5. Friedrich Möglich - sein Beitrag zum Aufbau der Physik in der DDR. Friedrich Möglich - langjähriger Mitherausgeber und Chefredakteur der Annalen der Physik

    NASA Astrophysics Data System (ADS)

    Rompe, Robert

    Friedrich Möglich, ein Schüler von Max von Laue und Erwin Schrödinger, übernahm 1947 die Chefredaktion der Annalen der Physik.Translated AbstractFriedrich Möglich - His Contributions to the Formation of Physics in GDRFriedrich Möglich a student of Max von Laue and Erwin Schrödinger took over as editor in chief of Annalen der Physik in 1947.

  6. Tycho Brahe - Instrumentenbauer und Meister der Beobachtungstechnik

    NASA Astrophysics Data System (ADS)

    Wolfschmidt, Gudrun

    Vor der Erfindung des Fernrohrs war der dänische Astronom Tycho Brahe (1546 - 1601) der bedeutendste beobachtende Astronom. Von seinem Observatorium Uraniborg auf der - damals dänischen - Insel Hven ist heute noch der Grundriß erkennbar, von Stjerneborg sind die Fundamente erhalten, die Kuppeln in den 1950er Jahren ergänzt. In der Astronomie-Ausstellung im Deutschen Museum gibt es ein Modell der Sternwarte Uraniborg und der zugehörigen Instrumente (Maßstab 1:10); das größere Modell wurde dem Technischen Museum in Malmö geschenkt. Die Instrumente, die er in den Observatorien Uraniborg und Stjerneborg benutzte, sind nicht erhalten. Aber es gibt gute Beschreibungen der Instrumente (Halbkreis, Quadranten, Sextanten, Armillarsphären, Triquetrum, Himmelsglobus) in seinem Buch Astronomiae instauratae mechanica (Wandsbek 1598). Eine Nachbildung des großen hölzernen Quadranten kann man im Runden Turm in Kopenhagen sehen. Zwei Sextanten, hergestellt für Tycho um 1600 von Jost Bürgi und Erasmus Habermel, gibt es noch im Nationalmuseum für Technik in Prag. Ähnlichkeiten von Tychos Instrumenten mit Groß-Instrumenten aus dem islamischen Kulturkreis sind auffällig. Tycho Brahes Meßgeräte markieren einen großen Fortschritt in der Entwicklung astronomischer Instrumente und Meßtechniken und bilden die Grundlage für den weiteren Fortschritt der Positionsastronomie und der damit verbundenen Tabellenwerke. Die Nachwirkungen sind bis ins 17. und 18. Jahrhundert nachweisbar.

  7. Untersuchung der Richtwirkung der Einkopplung von ebenen Wellen in eine Leitung

    NASA Astrophysics Data System (ADS)

    Magdowski, M.; Vick, R.

    2013-07-01

    Elektrische Leitungen und Kabel stellen häufig die Haupteinfallstore für elektromagnetische Felder in die daran angeschlossenen Geräte und Systeme dar. Für die Einkopplung einer ebenen Welle kann der in eine Leitung eingekoppelte Strom unter bestimmten Voraussetzungen mit Hilfe der Leitungstheorie sehr effizient bestimmt werden. Er hängt dabei von den Abmessungen der Leitung, den Leitungsabschlüssen sowie der Amplitude, der Wellenlänge und der Einfallsrichtung der ebenen Welle ab. In dieser Arbeit wird die Abhängigkeit der Einkopplung von der Einfallsrichtung näher untersucht. Dazu werden Richtdiagramme der Einkopplung berechnet, dargestellt und hinsichtlich der mittleren und maximalen Einkopplung über alle Einfallsrichtungen und Polarisationen ausgewertet. Die Ergebnisse werden genutzt, um die maximale Direktivität der Einkopplung in eine Leitung zu bestimmen. Fasst man die Einkopplung externer Felder in eine Leitung als einen Störfestigkeitstest auf, so kann die maximale Direktivität benutzt werden, um einen Vergleich zwischen unterschiedlichen Messumgebungen wie Absorberhallen und Modenverwirbelungskammern herzustellen.

  8. Von Humboldt bis Einstein. Berlin als Weltzentrum der exakten Wissenschaften.

    NASA Astrophysics Data System (ADS)

    Meschkowski, H.

    Contents: 1. Die Anfänge. 2. Die Ära Dirichlet-Jacobi. 3. Der Ausbau der experimentellen Naturwissenschaften. 4. Alexander von Humboldt. 5. Berlin wird "Weltzentrum" der Mathematik. 6. Die Ära Helmholtz. 7. Neue Arbeitsweisen der Astronomie. 8. Chemie: Forschung und Industrie. 9. Max Planck. 10. Ins technische Zeitalter. 11. Zur Mathematik der zwanziger Jahre. 12. Albert Einstein. 13. Fortschritte der Grundlagenforschung. 14. Erwin Schrödinger: Physiker, Philosoph und Poet. 15. Zum Schluß.

  9. Metabolic Flux Analysis of Lipid Biosynthesis in the Yeast Yarrowia lipolytica Using 13C-Labled Glucose and Gas Chromatography-Mass Spectrometry.

    PubMed

    Zhang, Huaiyuan; Wu, Chao; Wu, Qingyu; Dai, Junbiao; Song, Yuanda

    2016-01-01

    The oleaginous yeast Yarrowia lipolytica has considerable potential for producing single cell oil, which can be converted to biodiesel, a sustainable alternative to fossil fuels. However, extensive fundamental and engineering efforts must be carried out before commercialized production become cost-effective. Therefore, in this study, metabolic flux analysis of Y. lipolytica was performed using 13C-labeled glucose as a sole carbon source in nitrogen sufficient and insufficient media. The nitrogen limited medium inhibited cell growth while promoting lipid accumulation (from 8.7% of their biomass to 14.3%). Metabolic flux analysis showed that flux through the pentose phosphate pathway was not significantly regulated by nitrogen concentration, suggesting that NADPH generation is not the limiting factor for lipid accumulation in Y. lipolytica. Furthermore, metabolic flux through malic enzyme was undetectable, confirming its non-regulatory role in lipid accumulation in this yeast. Nitrogen limitation significantly increased flux through ATP:citrate lyase (ACL), implying that ACL plays a key role in providing acetyl-CoA for lipid accumulation in Y. lipolytica. PMID:27454589

  10. The fatty acid transport protein Fat1p is involved in the export of fatty acids from lipid bodies in Yarrowia lipolytica.

    PubMed

    Dulermo, Rémi; Gamboa-Meléndez, Heber; Dulermo, Thierry; Thevenieau, France; Nicaud, Jean-Marc

    2014-09-01

    In order to live, cells need to import different molecules, such as sugars, amino acids or lipids, using transporters. In Saccharomyces cerevisiae, the ScFAT1 gene encodes the long-chain fatty acid transporter; however, the transport of fatty acids (FAs) in the oleaginous yeast Yarrowia lipolytica has not yet been studied. In contrast to what has previously been found for ΔScfat1 strains, ΔYlfat1 yeast was still able to grow on substrates containing short-, medium- or long-chain FAs. We observed a notable difference in cell lipid content between wild-type (WT) and deletion mutant strains after 24 h of culture in minimal oleate medium: in the WT strain, lipids represented 24% of cell dry weight (CDW), while they accounted for 37% of CDW in the ΔYlfat1 strain. This result indicates that YlFat1p is not involved in cell lipid uptake. Moreover, we also observed that fatty acid remobilisation was decreased in the ΔYlfat1 strain and that fluorescence-tagged YlFat1p proteins localised to the interfaces between lipid bodies, which suggests that YlFat1p may play a role in the export of FAs from lipid bodies. PMID:24945074

  11. Citric acid production from partly deproteinized whey under non-sterile culture conditions using immobilized cells of lactose-positive and cold-adapted Yarrowia lipolytica B9.

    PubMed

    Arslan, Nazli Pinar; Aydogan, Mehmet Nuri; Taskin, Mesut

    2016-08-10

    The present study was performed to produce citric acid (CA) from partly deproteinized cheese whey (DPCW) under non-sterile culture conditions using immobilized cells of the cold-adapted and lactose-positive yeast Yarrowia lipolytica B9. DPCW was prepared using the temperature treatment of 90°C for 15min. Sodium alginate was used as entrapping agent for cell immobilization. Optimum conditions for the maximum CA production (33.3g/L) in non-sterile DPCW medium were the temperature of 20°C, pH 5.5, additional lactose concentration of 20g/L, sodium alginate concentration of 2%, number of 150 beads/100mL and incubation time of 120h. Similarly, maximum citric acid/isocitric acid (CA/ICA) ratio (6.79) could be reached under these optimal conditions. Additional nitrogen and phosphorus sources decreased CA concentration and CA/ICA ratio. Immobilized cells were reused in three continuous reaction cycles without any loss in the maximum CA concentration. The unique combination of low pH and temperature values as well as cell immobilization procedure could prevent undesired microbial contaminants during CA production. This is the first work on CA production by cold-adapted microorganisms under non-sterile culture conditions. Besides, CA production using a lactose-positive strain of the yeast Y. lipolytica was investigated for the first time in the present study. PMID:27234881

  12. Selection of new over-producing derivatives for the improvement of extracellular lipase production by the non-conventional yeast Yarrowia lipolytica.

    PubMed

    Fickers, Patrick; Fudalej, Franck; Nicaud, Jean-Marc; Destain, Jacqueline; Thonart, Phillipe

    2005-02-23

    The non-conventional yeast Yarrowia lipolytica produces an extracellular lipase encoded by the LIP2 gene. Mutant strains with enhanced productivity were previously obtained either by chemical mutagenesis or genetic engineering. In this work, we used one of these mutants, named LgX64.81 to select new overproducing strains following by amplification of the LIP2 gene. We also developed a process for lipase production in bioreactors and compared lipase production levels in batch and fed-batch cultures. Batch culture led to a lipase production of 26450 U ml(-1) in a media containing olive oil and tryptone as carbon and nitrogen sources. Feeding of a combination of tryptone and olive oil at the end of the exponential growth phase yielded to lipase activity of 158246 U ml(-1) after 80 h of cultivation. In addition this production system developed for the extracellular lipase could also be applied for other heterologous protein production since we have demonstrated that LgX64.81 is an interesting alternative host strain. PMID:15639099

  13. Yarrowia lipolytica Cells Mutant for the PEX24 Gene Encoding a Peroxisomal Membrane Peroxin Mislocalize Peroxisomal Proteins and Accumulate Membrane Structures Containing Both Peroxisomal Matrix and Membrane Proteins

    PubMed Central

    Tam, Yuen Yi C.; Rachubinski, Richard A.

    2002-01-01

    Peroxins are proteins required for peroxisome assembly and are encoded by the PEX genes. Functional complementation of the oleic acid–nonutilizing strain mut1-1 of the yeast Yarrowia lipolytica has identified the novel gene, PEX24. PEX24 encodes Pex24p, a protein of 550 amino acids (61,100 Da). Pex24p is an integral membrane protein of peroxisomes that exhibits high sequence homology to two hypothetical proteins encoded by the open reading frames YHR150W and YDR479C of the Saccharomyces cerevisiae genome. Pex24p is detectable in wild-type cells grown in glucose-containing medium, and its levels are significantly increased by incubation of cells in oleic acid–containing medium, the metabolism of which requires intact peroxisomes. pex24 mutants are compromised in the targeting of both matrix and membrane proteins to peroxisomes. Although pex24 mutants fail to assemble functional peroxisomes, they do harbor membrane structures that contain subsets of peroxisomal proteins. PMID:12181338

  14. Removal of hexavalent chromium ions by Yarrowia lipolytica cells modified with phyto-inspired Fe0/Fe3O4 nanoparticles

    NASA Astrophysics Data System (ADS)

    Rao, Ashit; Bankar, Ashok; Kumar, Ameeta Ravi; Gosavi, Suresh; Zinjarde, Smita

    2013-03-01

    The removal of hexavalent chromium [Cr (VI)], an important ground water pollutant by phyto-inspired Fe0/Fe3O4 nanocomposite-modified cells of Yarrowia lipolytica (NCIM 3589 and NCIM 3590), was investigated. Electron microscopy and magnetometer studies indicated an effective modification of yeast cell surfaces by the nanocomposites. The effect of pH, temperature, agitation speed, contact time and initial metal ion concentration on the removal of Cr (VI) was determined. The specific uptake values at pH 2.0 were 186.32 ± 3.17 and 137.31 ± 4.53 mg g- 1 for NCIM 3589 and NCIM 3590, respectively, when 1000 mg L- 1 of metal ion concentrations were used. The equilibrium data fitted to Scatchard, Langmuir and linearized Freundlich models suggesting that adsorption played a role in the removal of Cr (VI) ions. The surface modified yeast cells displayed higher values of Langmuir and Scatchard coefficients than the unmodified cells indicating that the former were more efficient in Cr (VI) removal. The enhanced detoxification of Cr (VI) ions by this composite material could be attributed to the reductive power of the Fe0/Fe3O4 nanocomposites as well the yeast cell surface functional groups.

  15. Citric acid production from extract of Jerusalem artichoke tubers by the genetically engineered yeast Yarrowia lipolytica strain 30 and purification of citric acid.

    PubMed

    Wang, Ling-Fei; Wang, Zhi-Peng; Liu, Xiao-Yan; Chi, Zhen-Ming

    2013-11-01

    In this study, citric acid production from extract of Jerusalem artichoke tubers by the genetically engineered yeast Yarrowia lipolytica strain 30 was investigated. After the compositions of the extract of Jerusalem artichoke tubers for citric acid production were optimized, the results showed that natural components of extract of Jerusalem artichoke tubers without addition of any other components were suitable for citric acid production by the yeast strain. During 10 L fermentation using the extract containing 84.3 g L(-1) total sugars, 68.3 g L(-1) citric acid was produced and the yield of citric acid was 0.91 g g(-1) within 336 h. At the end of the fermentation, 9.2 g L(-1) of residual total sugar and 2.1 g L(-1) of reducing sugar were left in the fermented medium. At the same time, citric acid in the supernatant of the culture was purified. It was found that 67.2 % of the citric acid in the supernatant of the culture was recovered and purity of citric acid in the crystal was 96 %. PMID:23584740

  16. Acidic phospholipid-independent interaction of Yas3p, an Opi1-family transcriptional repressor of Yarrowia lipolytica, with the endoplasmic reticulum.

    PubMed

    Kobayashi, Satoshi; Tezaki, Satoshi; Horiuchi, Hiroyuki; Fukuda, Ryouichi; Ohta, Akinori

    2015-12-01

    In the n-alkane-assimilating yeast Yarrowia lipolytica, the transcription of ALK1, encoding cytochrome P450, that catalyses n-alkane hydroxylation is activated by a complex composed of Yas1p and Yas2p via a promoter element, ARE1, in response to n-alkanes. An Opi1-family transcription factor, Yas3p, represses the transcription by binding to Yas2p in the nucleus when cultured in glucose-containing medium, but it is localized to the ER, presumably through interaction with acidic phospholipids, phosphatidic acid and/or phospho inositides, when cultured in n-alkane-containing medium. Here, to elucidate the mechanisms regulating the localization of Yas3p, point and deletion mutants of Yas3p were constructed and analysed. The substitution of Trp(360) and Cys(361) by Arg abrogated the localization of Yas3p to the ER and decreased ARE1-mediated transcriptional activation by n-alkane. A Yas3p truncation mutant consisting of residues 259-422 did not bind to acidic phospholipids, but it was localized to the ER in the presence of n-alkane, implying the acidic-phospholipid-independent recruitment of this mutant to the ER in response to n-alkane. The W360R and C361R substitutions in this truncation mutant abolished its localization to the ER. The results suggest that these residues are implicated in the acidic phospholipid-independent interaction of Yas3p to the ER. PMID:26284565

  17. Analysis of ATP-citrate lyase and malic enzyme mutants of Yarrowia lipolytica points out the importance of mannitol metabolism in fatty acid synthesis.

    PubMed

    Dulermo, Thierry; Lazar, Zbigniew; Dulermo, Rémi; Rakicka, Magdalena; Haddouche, Ramedane; Nicaud, Jean-Marc

    2015-09-01

    The role of the two key enzymes of fatty acid (FA) synthesis, ATP-citrate lyase (Acl) and malic enzyme (Mae), was analyzed in the oleaginous yeast Yarrowia lipolytica. In most oleaginous yeasts, Acl and Mae are proposed to provide, respectively, acetyl-CoA and NADPH for FA synthesis. Acl was mainly studied at the biochemical level but no strain depleted for this enzyme was analyzed in oleaginous microorganisms. On the other hand the role of Mae in FA synthesis in Y. lipolytica remains unclear since it was proposed to be a mitochondrial NAD(H)-dependent enzyme and not a cytosolic NADP(H)-dependent enzyme. In this study, we analyzed for the first time strains inactivated for corresponding genes. Inactivation of ACL1 decreases FA synthesis by 60 to 80%, confirming its essential role in FA synthesis in Y. lipolytica. Conversely, inactivation of MAE1 has no effects on FA synthesis, except in a FA overaccumulating strain where it improves FA synthesis by 35%. This result definitively excludes Mae as a major key enzyme for FA synthesis in Y. lipolytica. During the analysis of both mutants, we observed a negative correlation between FA and mannitol level. As mannitol and FA pathways may compete for carbon storage, we inactivated YlSDR, encoding a mannitol dehydrogenase converting fructose and NADPH into mannitol and NADP+. The FA content of the resulting mutant was improved by 60% during growth on fructose, demonstrating that mannitol metabolism may modulate FA synthesis in Y. lipolytica. PMID:25959598

  18. Involvement of acyl-CoA synthetase genes in n-alkane assimilation and fatty acid utilization in yeast Yarrowia lipolytica.

    PubMed

    Tenagy; Park, Jun Seok; Iwama, Ryo; Kobayashi, Satoshi; Ohta, Akinori; Horiuchi, Hiroyuki; Fukuda, Ryouichi

    2015-06-01

    Here, we investigated the roles of YAL1 (FAA1) and FAT1 encoding acyl-CoA synthetases (ACSs) and three additional orthologs of ACS genes FAT2-FAT4 of the yeast Yarrowia lipolytica in the assimilation or utilization of n-alkanes and fatty acids. ACS deletion mutants were generated to characterize their function. The FAT1 deletion mutant exhibited decreased growth on n-alkanes of 10-18 carbons, whereas the FAA1 mutant showed growth reduction on n-alkane of 16 carbons. However, FAT2-FAT4 deletion mutants did not show any growth defects, suggesting that FAT1 and FAA1 are involved in the activation of fatty acids produced during the metabolism of n-alkanes. In contrast, deletions of FAA1 and FAT1-FAT4 conferred no defect in growth on fatty acids. The wild-type strain grew in the presence of cerulenin, an inhibitor of fatty acid synthesis, by utilizing exogenously added fatty acid or fatty acid derived from n-alkane when oleic acid or n-alkane of 18 carbons was supplemented. However, the FAA1 deletion mutant did not grow, indicating a critical role for FAA1 in the utilization of fatty acids. Fluorescent microscopic observation and biochemical analyses suggested that Fat1p is present in the peroxisome and Faa1p is localized in the cytosol and to membranes. PMID:26019148

  19. Optimization of a low-cost hyperosmotic medium and establishing the fermentation kinetics of erythritol production by Yarrowia lipolytica from crude glycerol.

    PubMed

    Yang, Li-Bo; Zhan, Xiao-Bei; Zhu, Li; Gao, Min-Jie; Lin, Chi-Chung

    2016-05-18

    The production of erythritol by Yarrowia lipolytica from low-cost substitutable substrates for high yield was investigated. Crude glycerol, urea, and NaCl related to osmotic pressure were the most significant factors affecting erythritol production. An artificial neural network model and genetic algorithm were used to search the optimal composition of the significant factors and locate the resulting erythritol yield. Medium with 232.39 g/L crude glycerol, 1.57 g/L urea, and 31.03 g/L NaCl led to predictive maximum erythritol concentration of 110.7 g/L. The erythritol concentration improved from 50.4 g/L to 109.2 g/L with the optimized medium, which was reproducible. Erythritol fermentation kinetics were investigated in a batch system. Multistep fermentation kinetic models with hyperosmotic inhibitory effects were developed. The resulting mathematical equations provided a good description of temporal variations such as microbial growth (X), substrate consumption (S), and product formation (P) in erythritol fermentation. The accordingly derived model is the first reported model for fermentative erythritol production from glycerol, providing useful information to optimize the growth of Y. lipolytica and contributing visual description for the erythritol fermentation process under high osmotic pressure, as well as improvement of productivity and efficiency. PMID:26176584

  20. Efficient biosynthesis of γ-decalactone in ionic liquids by immobilized whole cells of Yarrowia lipolytica G3-3.21 on attapulgite.

    PubMed

    Zhao, Yuping; Xu, Yan; Jiang, Changxing

    2015-10-01

    In this study, the biosynthesis of γ-decalactone (GDL) was successfully conducted in an ionic liquid (IL)-containing cosolvent system using immobilized cells of Yarrowia lipolytica G3-3.21 on attapulgite (ATG). We found the immobilized Y. lipolytica G3-3.21 cells in N-butyl-pyridinium tetrafluoroborate ([BPy]BF4) solution gave the highest activity of C16-Acyl-CoA oxidase and the maximum yield of GDL. The optimum immobilization conditions for the highest yield of GDL were 20 g/L of ATG, 1.5 % of CaCl2 and 2 % of sodium alginate (NaAlg). The optimal [BPy]BF4 content, buffer pH, reaction temperature, shaking speed, castor oil and glucose contents were 7.5 %, 26 °C, 150 rpm, 100 g/L and 10 %, respectively. Under the optimized conditions, the GDL yield was up to 8.05 g/L. After ten times of reuse, the GDL yield was 7.51 g/L, corresponding to 93.3 % of that obtained in the first batch, suggesting a good reusability and potential for industrial applications. PMID:26091898

  1. L-Phenylalanine catabolism and 2-phenylethanol synthesis in Yarrowia lipolytica--mapping molecular identities through whole-proteome quantitative mass spectrometry analysis.

    PubMed

    Celińska, Ewelina; Olkowicz, Mariola; Grajek, Włodzimierz

    2015-08-01

    A world-wide effort is now being pursued towards the development of flavors and fragrances (F&F) production independently from traditional sources, as well as autonomously from depleting fossil fuel supplies. Biotechnological production of F&F by microbes has emerged as a vivid solution to the current market limitations. Amongst a wide variety of fragrant chemicals, 2-PE is of significant interest to both scientific and industrial community. Although the general overview of the 2-PE synthesis pathway is commonly known, involvement of particular molecular identities in this pathway has not been elucidated in Yarrowia lipolytica to date. The aim of this study was mapping molecular identities involved in 2-PE synthesis in Y. lipolytica. To acquire a comprehensive landscape of the proteins that are directly and indirectly involved in L-Phe degradation and 2-PE synthesis, we took advantage of comprehensibility and sensitivity of high-throughput LC-MS/MS-quantitative analysis. Amongst a number of proteins involved in amino acid turnover and the central carbon metabolism, enzymes involved in L-Phe conversion to 2-PE have been identified. Results on yeast-to-hyphae transition in relation to the character of the provided nitrogen source have been presented. PMID:26060219

  2. A newly identified fatty alcohol oxidase gene is mainly responsible for the oxidation of long-chain ω-hydroxy fatty acids in Yarrowia lipolytica.

    PubMed

    Gatter, Michael; Förster, André; Bär, Kati; Winter, Miriam; Otto, Christina; Petzsch, Patrick; Ježková, Michaela; Bahr, Katrin; Pfeiffer, Melanie; Matthäus, Falk; Barth, Gerold

    2014-09-01

    Nine potential (fatty) alcohol dehydrogenase genes and one alcohol oxidase gene were identified in Yarrowia lipolytica by comparative sequence analysis. All relevant genes were deleted in Y. lipolytica H222ΔP which is lacking β-oxidation. Resulting transformants were tested for their ability to accumulate ω-hydroxy fatty acids and dicarboxylic acids in the culture medium. The deletion of eight alcohol dehydrogenase genes (FADH, ADH1-7), which may be involved in ω-oxidation, led only to a slightly increased accumulation of ω-hydroxy fatty acids, whereas the deletion of the fatty alcohol oxidase gene (FAO1), which has not been described yet in Y. lipolytica, exhibited a considerably higher effect. The combined deletion of the eight (fatty) alcohol dehydrogenase genes and the alcohol oxidase gene further reduced the formation of dicarboxylic acids. These results indicate that both (fatty) alcohol dehydrogenases and an alcohol oxidase are involved in ω-oxidation of long-chain fatty acids whereby latter plays the major role. This insight marks the first step toward the biotechnological production of long-chain ω-hydroxy fatty acids with the help of the nonconventional yeast Y. lipolytica. The overexpression of FAO1 can be further used to improve existing strains for the production of dicarboxylic acids. PMID:24931727

  3. Improved production of fatty acids by Saccharomyces cerevisiae through screening a cDNA library from the oleaginous yeast Yarrowia lipolytica.

    PubMed

    Shi, Shuobo; Ji, Haichuan; Siewers, Verena; Nielsen, Jens

    2016-02-01

    Biological production of fatty acid (FA)-derived products has gained increasing attention to replace petroleum-based fuels and chemicals. FA biosynthesis is highly regulated, and usually it is challenging to design rational engineering strategies. In addition, the conventional 'one sample at a time' method for lipid determination is time consuming and laborious, and it is difficult to screen large numbers of samples. Here, a method for detecting free FAs in viable cells using Nile red staining was developed for use in large-scale screening. Following optimization of the method, it was used for screening a cDNA library from the oleaginous yeast Yarrowia lipolytica for identification of genes/enzymes that were able to enhance free FA accumulation in Saccharomyces cerevisiae. Several novel enzymes resulting in increasing FA accumulation were discovered. These targets include a GPI anchor protein, malate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, FA hydroxylase, farnesyltransferase, anoctamin, dihydrolipoamide dehydrogenase and phosphatidylethanolamine-binding protein. The best enzyme resulted in a 2.5-fold improvement in production of free FAs. Our findings not only provide a novel method for high-throughput evaluation of the content of free FAs, but also give new insight into how enzymes from Y. lipolytica may increase the production of fatty acids in S. cerevisiae. PMID:26658002

  4. Extracellular expression of YlLip11 with a native signal peptide from Yarrowia lipolytica MSR80 in three different yeast hosts.

    PubMed

    Kumari, Arti; Baronian, Keith; Kunze, Gotthard; Gupta, Rani

    2015-06-01

    Lipase YlLip11 from Yarrowia lipolytica was expressed with a signal peptide encoding sequence in Arxula adeninivorans, Saccharomyces cerevisiae and Hansenula polymorpha using the Xplor®2 transformation/expression platform and an expression module with the constitutive Arxula-derived TEF1 promoter. The YlLip11 signal peptide was functional in all of the yeast hosts with 97% of the recombinant enzyme being secreted into the culture medium. However, recombinant YlLip11 with His Tag fused at C-terminal was not active. The best recombinant YlLip11 producing A. adeninivorans G1212/YRC102-YlLip11 transformant cultivated in shake flasks produced 2654 U/L lipase, followed by S. cerevisiae SEY6210/YRC103-YlLip11 (1632U/L) and H. polymorpha RB11/YRC103-YlLip11 (1144U/L). Although the biochemical parameters of YlLip11 synthesized in different hosts were similar, their glycosylation level and thermo stability differed. The protein synthesized by the H. polymorpha transformant had the highest degree of glycosylation and with a t1/2 of 60min at 70°C, exhibited the highest thermostability. PMID:25725269

  5. Metabolic Flux Analysis of Lipid Biosynthesis in the Yeast Yarrowia lipolytica Using 13C-Labled Glucose and Gas Chromatography-Mass Spectrometry

    PubMed Central

    Zhang, Huaiyuan; Wu, Chao; Wu, Qingyu; Dai, Junbiao; Song, Yuanda

    2016-01-01

    The oleaginous yeast Yarrowia lipolytica has considerable potential for producing single cell oil, which can be converted to biodiesel, a sustainable alternative to fossil fuels. However, extensive fundamental and engineering efforts must be carried out before commercialized production become cost-effective. Therefore, in this study, metabolic flux analysis of Y. lipolytica was performed using 13C-labeled glucose as a sole carbon source in nitrogen sufficient and insufficient media. The nitrogen limited medium inhibited cell growth while promoting lipid accumulation (from 8.7% of their biomass to 14.3%). Metabolic flux analysis showed that flux through the pentose phosphate pathway was not significantly regulated by nitrogen concentration, suggesting that NADPH generation is not the limiting factor for lipid accumulation in Y. lipolytica. Furthermore, metabolic flux through malic enzyme was undetectable, confirming its non-regulatory role in lipid accumulation in this yeast. Nitrogen limitation significantly increased flux through ATP:citrate lyase (ACL), implying that ACL plays a key role in providing acetyl-CoA for lipid accumulation in Y. lipolytica. PMID:27454589

  6. Role of surface-inoculated Debaryomyces hansenii and Yarrowia lipolytica strains in dried fermented sausage manufacture. Part 2: Evaluation of their effects on sensory quality and biogenic amine content.

    PubMed

    Iucci, Luciana; Patrignani, Francesca; Belletti, Nicoletta; Ndagijimana, Maurice; Elisabetta Guerzoni, M; Gardini, Fausto; Lanciotti, Rosalba

    2007-04-01

    The aim was to study the effects of Debaryomyces hansenii and Yarrowia lipolytica strains, used with lactic acid starter cultures (Lactobacillus plantarum), in the manufacture of dried fermented sausages in order to understand their effects on volatile profile, biogenic amine content and sensory properties. The experimental data showed that every yeast strain produced a specific profile of volatile metabolic products. The yeasts also gave sausages with distinctive sensory properties. The degree of mincing also influenced these properties, but none of these factors had significant influence upon the accumulation of biogenic amines. PMID:22064032

  7. Use of Plackett-Burman design for rapid screening of nitrogen and carbon sources for the production of lipase in solid state fermentation by Yarrowia lipolytica from mustard oil cake (Brassica napus)

    PubMed Central

    Imandi, Sarat Babu; Karanam, Sita Kumari; Garapati, Hanumantha Rao

    2013-01-01

    Mustard oil cake (Brassica napus), the residue obtained after extraction of mustard oil from mustard oil seeds, was investigated for the production of lipase under solid state fermentation (SSF) using the marine yeast Yarrowia lipolytica NCIM 3589. Process parameters such as incubation time, biomass concentration, initial moisture content, carbon source concentration and nitrogen source concentration of the medium were optimized. Screening of ten nitrogen and five carbon sources has been accomplished with the help of Plackett-Burman design. The highest lipase activity of 57.89 units per gram of dry fermented substrate (U/gds) was observed with the substrate of mustard oil cake in four days of fermentation. PMID:24516460

  8. Use of Plackett-Burman design for rapid screening of nitrogen and carbon sources for the production of lipase in solid state fermentation by Yarrowia lipolytica from mustard oil cake (Brassica napus).

    PubMed

    Imandi, Sarat Babu; Karanam, Sita Kumari; Garapati, Hanumantha Rao

    2013-01-01

    Mustard oil cake (Brassica napus), the residue obtained after extraction of mustard oil from mustard oil seeds, was investigated for the production of lipase under solid state fermentation (SSF) using the marine yeast Yarrowia lipolytica NCIM 3589. Process parameters such as incubation time, biomass concentration, initial moisture content, carbon source concentration and nitrogen source concentration of the medium were optimized. Screening of ten nitrogen and five carbon sources has been accomplished with the help of Plackett-Burman design. The highest lipase activity of 57.89 units per gram of dry fermented substrate (U/gds) was observed with the substrate of mustard oil cake in four days of fermentation. PMID:24516460

  9. Werner Heisenberg zum 100. Geburtstag: Pionier der Quantenmechanik

    NASA Astrophysics Data System (ADS)

    Jacobi, Manfred

    2001-11-01

    Werner Heisenberg war eine der prägendsten Gestalten der Physik des 20. Jahrhunderts. Zu seinen wichtigsten Verdiensten gehören die Grundlegung der Quantenmechanik, die Formulierung der Unschärferelationen sowie die Beteiligung an der Ausarbeitung der Kopenhagener Deutung der Quantenmechanik. Darüber hinaus lieferte er Arbeiten von fundamentalem Charakter zur Theorie des Atomkerns, zur kosmischen Strahlung und zur Quantenfeldtheorie. Während des Krieges war er an den Arbeiten des Uranvereins beteiligt, der die Möglichkeit einer Entwicklung von Kernwaffen untersuchte, jedoch über Vorarbeiten zur Reaktorphysik nicht hinauskam. Wegen dieser Tätigkeit wurde er bei Kriegsende für einige Monate in England interniert. Nach seiner Rückkehr widmete er sich vor allem dem Aufbau der Physik in Deutschland, die während der NS-Zeit nahezu ihrer gesamten Substanz beraubt worden war.

  10. ARPES studies of van der Waals heterostructure

    NASA Astrophysics Data System (ADS)

    Wang, Eryin; Lu, Xiaobo; Chen, Guorui; Fedorov, Alexei V.; Zhang, Yuanbo; Zhang, Guangyu; Zhou, Shuyun

    Van der Waals heterostructures are a novel class of ``materials by design'' which are formed by stacking different two-dimensional crystals together via van der Waals interaction. The periodic potential by the Moir é superlattice can be used as a control knob for tuning the electronic properties of two dimensional materials and can induce various novel quantum phenomena. Here we report direct electronic structure studies the of a model van der Waals heterostructure using angle-resolved photoemission spectroscopy (ARPES). This work is supported by the National Natural Science Foundation of China and Ministry of Education of China.

  11. Qualitätsmanagement in der Lebensmittelindustrie

    NASA Astrophysics Data System (ADS)

    Thorn, Volker

    Die wesentlichen Kunden der Lebensmittelindustrie sind der Einzel- und Großhandel und die Verbraucher. Jedes Unternehmen kann mittel- und langfristig nur existieren, wenn seine Kunden zufrieden sind. Kunden sind zufrieden, wenn ihre Erwartungen, die sie an Produkt, Service und Preis stellen, erfüllt werden. Also die bestimmte erwartete Qualität (Leistung) sichergestellt wird. Trotz aller Bemühungen und Anstrengungen der Anbieter, Qualitätsprodukte auf den Markt zu bringen, kames in den letzten Jahren immer wieder zu Lebensmittelskandalen.

  12. Molecular Characterization of the Elaeis guineensis Medium-Chain Fatty Acid Diacylglycerol Acyltransferase DGAT1-1 by Heterologous Expression in Yarrowia lipolytica.

    PubMed

    Aymé, Laure; Jolivet, Pascale; Nicaud, Jean-Marc; Chardot, Thierry

    2015-01-01

    Diacylglycerol acyltransferases (DGAT) are involved in the acylation of sn-1,2-diacylglycerol. Palm kernel oil, extracted from Elaeis guineensis (oil palm) seeds, has a high content of medium-chain fatty acids mainly lauric acid (C12:0). A putative E. guineensis diacylglycerol acyltransferase gene (EgDGAT1-1) is expressed at the onset of lauric acid accumulation in the seed endosperm suggesting that it is a determinant of medium-chain triacylglycerol storage. To test this hypothesis, we thoroughly characterized EgDGAT1-1 activity through functional complementation of a Yarrowia lipolytica mutant strain devoid of neutral lipids. EgDGAT1-1 expression is sufficient to restore triacylglycerol accumulation in neosynthesized lipid droplets. A comparative functional study with Arabidopsis thaliana DGAT1 highlighted contrasting substrate specificities when the recombinant yeast was cultured in lauric acid supplemented medium. The EgDGAT1-1 expressing strain preferentially accumulated medium-chain triacylglycerols whereas AtDGAT1 expression induced long-chain triacylglycerol storage in Y. lipolytica. EgDGAT1-1 localized to the endoplasmic reticulum where TAG biosynthesis takes place. Reestablishing neutral lipid accumulation in the Y. lipolytica mutant strain did not induce major reorganization of the yeast microsomal proteome. Overall, our findings demonstrate that EgDGAT1-1 is an endoplasmic reticulum DGAT with preference for medium-chain fatty acid substrates, in line with its physiological role in palm kernel. The characterized EgDGAT1-1 could be used to promote medium-chain triacylglycerol accumulation in microbial-produced oil for industrial chemicals and cosmetics. PMID:26581109

  13. Fe(III) mineral reduction followed by partial dissolution and reactive oxygen species generation during 2,4,6-trinitrotoluene transformation by the aerobic yeast Yarrowia lipolytica.

    PubMed

    Ziganshin, Ayrat M; Ziganshina, Elvira E; Byrne, James; Gerlach, Robin; Struve, Ellen; Biktagirov, Timur; Rodionov, Alexander; Kappler, Andreas

    2015-01-01

    Understanding the factors that influence pollutant transformation in the presence of ferric (oxyhydr)oxides is crucial to the efficient application of different remediation strategies. In this study we determined the effect of goethite, hematite, magnetite and ferrihydrite on the transformation of 2,4,6-trinitrotoluene (TNT) by Yarrowia lipolytica AN-L15. The presence of ferric (oxyhydr)oxides led to a small decrease in the rate of TNT removal. In all cases, a significant release of NO2 (-) from TNT and further NO2 (-) oxidation to NO3 (-) was observed. A fraction of the released NO2 (-) was abiotically decomposed to NO and NO2, and then NO was likely oxidized abiotically to NO2 by O2. ESR analysis revealed the generation of superoxide in the culture medium; its further protonation at low pH resulted in the formation of hydroperoxyl radical. Presumably, a fraction of NO released during TNT degradation reacted with superoxide and formed peroxynitrite, which was further rearranged to NO3 (-) at the acidic pH values observed in this study. A transformation and reduction of ferric (oxyhydr)oxides followed by partial dissolution (in the range of 7-86% of the initial Fe(III)) were observed in the presence of cells and TNT. Mössbauer spectroscopy showed some minor changes for goethite, magnetite and ferrihydrite samples during their incubation with Y. lipolytica and TNT. This study shows that i) reactive oxygen and nitrogen species generated during TNT transformation by Y. lipolytica participate in the abiotic conversion of TNT and ii) the presence of iron(III) minerals leads to a minor decrease in TNT transformation. PMID:25852985

  14. Molecular Characterization of the Elaeis guineensis Medium-Chain Fatty Acid Diacylglycerol Acyltransferase DGAT1-1 by Heterologous Expression in Yarrowia lipolytica

    PubMed Central

    Aymé, Laure; Jolivet, Pascale; Nicaud, Jean-Marc; Chardot, Thierry

    2015-01-01

    Diacylglycerol acyltransferases (DGAT) are involved in the acylation of sn-1,2-diacylglycerol. Palm kernel oil, extracted from Elaeis guineensis (oil palm) seeds, has a high content of medium-chain fatty acids mainly lauric acid (C12:0). A putative E. guineensis diacylglycerol acyltransferase gene (EgDGAT1-1) is expressed at the onset of lauric acid accumulation in the seed endosperm suggesting that it is a determinant of medium-chain triacylglycerol storage. To test this hypothesis, we thoroughly characterized EgDGAT1-1 activity through functional complementation of a Yarrowia lipolytica mutant strain devoid of neutral lipids. EgDGAT1-1 expression is sufficient to restore triacylglycerol accumulation in neosynthesized lipid droplets. A comparative functional study with Arabidopsis thaliana DGAT1 highlighted contrasting substrate specificities when the recombinant yeast was cultured in lauric acid supplemented medium. The EgDGAT1-1 expressing strain preferentially accumulated medium-chain triacylglycerols whereas AtDGAT1 expression induced long-chain triacylglycerol storage in Y. lipolytica. EgDGAT1-1 localized to the endoplasmic reticulum where TAG biosynthesis takes place. Reestablishing neutral lipid accumulation in the Y. lipolytica mutant strain did not induce major reorganization of the yeast microsomal proteome. Overall, our findings demonstrate that EgDGAT1-1 is an endoplasmic reticulum DGAT with preference for medium-chain fatty acid substrates, in line with its physiological role in palm kernel. The characterized EgDGAT1-1 could be used to promote medium-chain triacylglycerol accumulation in microbial-produced oil for industrial chemicals and cosmetics. PMID:26581109

  15. Cloning and Characterization of a Pyruvate Carboxylase Gene from Penicillium rubens and Overexpression of the Genein the Yeast Yarrowia lipolytica for Enhanced Citric Acid Production.

    PubMed

    Fu, Ge-Yi; Lu, Yi; Chi, Zhe; Liu, Guang-Lei; Zhao, Shou-Feng; Jiang, Hong; Chi, Zhen-Ming

    2016-02-01

    In this study, a pyruvate carboxylase gene (PYC1) from a marine fungus Penicillium rubens I607 was cloned and characterized. ORF of the gene (accession number: KM397349.1) had 3534 bp encoding 1177 amino acids with a molecular weight of 127.531 kDa and a PI of 6.20. The promoter of the gene was located at -1200 bp and contained a TATAA box, several CAAT boxes and a sequence 5'-SYGGRG-3'. The PYC1 deduced from the gene had no signal peptide, was a homotetramer (α4), and had the four functional domains. After expression of the PYC1 gene from the marine fungus in the marine-derived yeast Yarrowia lipolytica SWJ-1b, the transformant PR32 obtained had much higher specific pyruvate carboxylase activity (0.53 U/mg) than Y. lipolytica SWJ-1b (0.07 U/mg), and the PYC1 gene expression (133.8%) and citric acid production (70.2 g/l) by the transformant PR32 were also greatly enhanced compared to those (100 % and 27.3 g/l) by Y. lipolytica SWJ-1b. When glucose concentration in the medium was 60.0 g/l, citric acid (CA) concentration formed by the transformant PR32 was 36.1 g/l, leading to conversion of 62.1% of glucose into CA. During a 10-l fed-batch fermentation, the final concentration of CA was 111.1 ± 1.3 g/l, the yield was 0.93 g/g, the productivity was 0.46 g/l/h, and only 1.72 g/l reducing sugar was left in the fermented medium within 240 h. HPLC analysis showed that most of the fermentation products were CA. However, minor malic acid and other unknown products also existed in the culture. PMID:26470708

  16. Functional roles and substrate specificities of twelve cytochromes P450 belonging to CYP52 family in n-alkane assimilating yeast Yarrowia lipolytica.

    PubMed

    Iwama, Ryo; Kobayashi, Satoshi; Ishimaru, Chiaki; Ohta, Akinori; Horiuchi, Hiroyuki; Fukuda, Ryouichi

    2016-06-01

    Yarrowia lipolytica possesses twelve ALK genes, which encode cytochromes P450 in the CYP52 family. In this study, using a Y. lipolytica strain from which all twelve ALK genes had been deleted, strains individually expressing each of the ALK genes were constructed and their roles and substrate specificities were determined by observing their growth on n-alkanes and analyzing fatty acid metabolism. The results suggested that the twelve Alk proteins can be categorized into four groups based on their substrate specificity: Alk1p, Alk2p, Alk9p, and Alk10p, which have significant activities to hydroxylate n-alkanes; Alk4p, Alk5p, and Alk7p, which have significant activities to hydroxylate the ω-terminal end of dodecanoic acid; Alk3p and Alk6p, which have significant activities to hydroxylate both n-alkanes and dodecanoic acid; and Alk8p, Alk11p, and Alk12p, which showed faint or no activities to oxidize these substrates. The involvement of Alk proteins in the oxidation of fatty alcohols and fatty aldehydes was also analyzed by measuring viability of the mutant deleted for twelve ALK genes in medium containing dodecanol and by observing growth on dodecanal of a mutant strain, in which twelve ALK genes were deleted along with four fatty aldehyde dehydrogenase genes. It was suggested that ALK gene(s) is/are involved in the detoxification of dodecanol and the assimilation of dodecanal. These results imply that genes encoding CYP52-family P450s have undergone multiplication and diversification in Y. lipolytica for assimilation of various hydrophobic compounds. PMID:27039152

  17. Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates.

    PubMed

    Lindquist, Mitch R; López-Núñez, Juan Carlos; Jones, Marjorie A; Cox, Elby J; Pinkelman, Rebecca J; Bang, Sookie S; Moser, Bryan R; Jackson, Michael A; Iten, Loren B; Kurtzman, Cletus P; Bischoff, Kenneth M; Liu, Siqing; Qureshi, Nasib; Tasaki, Kenneth; Rich, Joseph O; Cotta, Michael A; Saha, Badal C; Hughes, Stephen R

    2015-11-01

    Increased interest in sustainable production of renewable diesel and other valuable bioproducts is redoubling efforts to improve economic feasibility of microbial-based oil production. Yarrowia lipolytica is capable of employing a wide variety of substrates to produce oil and valuable co-products. We irradiated Y. lipolytica NRRL YB-567 with UV-C to enhance ammonia (for fertilizer) and lipid (for biodiesel) production on low-cost protein and carbohydrate substrates. The resulting strains were screened for ammonia and oil production using color intensity of indicators on plate assays. Seven mutant strains were selected (based on ammonia assay) and further evaluated for growth rate, ammonia and oil production, soluble protein content, and morphology when grown on liver infusion medium (without sugars), and for growth on various substrates. Strains were identified among these mutants that had a faster doubling time, produced higher maximum ammonia levels (enzyme assay) and more oil (Sudan Black assay), and had higher maximum soluble protein levels (Bradford assay) than wild type. When grown on plates with substrates of interest, all mutant strains showed similar results aerobically to wild-type strain. The mutant strain with the highest oil production and the fastest doubling time was evaluated on coffee waste medium. On this medium, the strain produced 0.12 g/L ammonia and 0.20 g/L 2-phenylethanol, a valuable fragrance/flavoring, in addition to acylglycerols (oil) containing predominantly C16 and C18 residues. These mutant strains will be investigated further for potential application in commercial biodiesel production. PMID:26272089

  18. Cell-surface expression of Aspergillus saitoi-derived functional α-1,2-mannosidase on Yarrowia lipolytica for glycan remodeling.

    PubMed

    Moon, Hye Yun; Van, Trinh Luu; Cheon, Seon Ah; Choo, Jinho; Kim, Jeong-Yoon; Kang, Hyun Ah

    2013-08-01

    Expression of proteins on the surface of yeast has a wide range of applications, such as development of live vaccines, screening of antibody libraries, and use as whole-cell biocatalysts. The hemiascomycetes yeast Yarrowia lipolytica has been raised as a potential host for heterologous expression of recombinant proteins. In this study, we report the expression of Aspergillus saitoi α-1,2-mannosidase, encoded by the msdS gene, on the cell surface of Y. lipolytica. As the first step to achieve the secretory expression of msdS protein, four different signal sequences-derived from the endogenous Y. lipolytica Lip2 and Xpr2 prepro regions and the heterologous A. niger α-amylase and rice α-amylase signal sequences-were analyzed for their secretion efficiency. It was shown that the YlLip2 prepro sequence was most efficient in directing the secretory expression of msdS in fully N-glycosylated forms. The surface display of msdS was subsequently directed by fusing GPI anchoring motifs derived from Y. lipolytica cell wall proteins, YlCwp1p and YlYwp1p, respectively, to the C-terminus of the Lip2 prepro-msdS protein. The expression of actively functional msdS protein on the cell surface was confirmed by western blot, flow cytometry analysis, along with the α-1,2-mannosidase activity assay using intact Y. lipolytica cells as the enzyme source. Furthermore, the glycoengineered Y. lipolytica Δoch1Δmpo1 strains displaying α-1,2-mannosidase were able to convert Man8GlcNAc2 to Man5GlcNAc2 efficiently on their cell-wall mannoproteins, demonstrating its potential used for glycoengineering in vitro or in vivo. PMID:23990303

  19. Genetics Home Reference: van der Woude syndrome

    MedlinePlus

    ... people with this disorder are born with a cleft lip , a cleft palate (an opening in the roof ... People with van der Woude syndrome who have cleft lip and/or palate, like other individuals with these ...

  20. Wie entsteht Bewusstsein?: Einfache Modelle der Selbstwahrnehmung

    NASA Astrophysics Data System (ADS)

    Schuster, Heinz Georg

    2003-07-01

    Was ist die funktionale Basis der Entstehung des Bewusstseins? Warum bringt dieses in der Evolution einen Vorteil? Diese Fragen werden anhand von drei Spielen untersucht, in denen die Spieler mathematisch modelliert sind. Im ersten Spiel lernt ein Spieler, seine Stellung in einer Hierarchie aus der Interaktion mit anderen zu erraten. Im zweiten Spiel macht ein Zuwachs des Gedächtnisses einen Spieler zum Gewinner: Das ermöglicht ihm einen Spiel entscheidenden Überblick über sein eigenes Verhalten und das seines Gegners. Im dritten Spiel müssen die Spieler die Farbe des Hutes auf dem eigenen Kopf aus der Reaktion des Gegners erraten. Hier ist die Fähigkeit von Vorteil, Inhalte aus dem Langzeit- und Kurzzeitgedächtnis im Geist abbilden und manipulieren zu können.

  1. Checkpoint-Inhibitoren in der Immuntherapie: Ein Meilenstein in der Behandlung des malignen Melanoms.

    PubMed

    Wilden, Sophia M; Lang, Berenice M; Mohr, Peter; Grabbe, Stephan

    2016-07-01

    Seit Jahrzehnten ist bekannt, dass Tumoren vom Immunsystem erkannt und zerstört werden können. Diese, vor allem in Tierversuchen gewonnene Erkenntnis konnte jedoch in der Vergangenheit nicht zum Nutzen unserer Patienten umgesetzt werden, da immunonkologische Therapieansätze in den letzten Jahrzehnten in der Anwendung beim Menschen stets versagt haben. Daher hat, mit Ausnahme der adjuvanten Interferontherapie, keines dieser Verfahren den Einzug in die klinische Versorgung gefunden. Langzeitüberleben unter guter Lebensqualität war dabei sehr wenigen Patienten vorbehalten. Mit den neuen immunologischen Therapieansätzen wird jedoch sowohl das Langzeitüberleben als auch die Lebensqualität onkologischer Patienten neu definiert. Auf die neuen "Immun-Checkpoint-Inhibitoren" spricht erstmals ein relevanter Teil der behandelten Patienten an und diese zeigen in der Regel langandauernde Remissionen bis hin zur Heilung. Schon jetzt ist klar, dass die Immuntherapie in Zukunft eine der wesentlichen Therapiesäulen bei der Behandlung des metastasierten Melanoms und auch vieler anderer fortgeschrittener Tumoren bilden wird. In dieser Übersicht werden die wichtigsten neuen Therapiemodalitäten besprochen und sowohl deren Wirkprinzip als auch klinische Daten zum Therapieansprechen und zu erwartenden Nebenwirkungen der Therapie referiert. PMID:27373243

  2. Wie wissenschaftlich ist der Evolutionsgedanke?

    NASA Astrophysics Data System (ADS)

    Vollmer, Gerhard

    Darwin war ein besonnener Mann; alles Aufsehen war ihm zuwider. Trotzdem hat er eine Revolution ausgelöst, deren Wirkung nicht auf die Biologie beschränkt blieb. Seine Theorie lässt sich in fünf Teiltheorien zerlegen, die sich durch die Begriffe Artenwandel, Verwandtschaft alles Lebendigen und gemeinsamer Ursprung, Artenaufspaltung und Artenvielfalt, Gradualismus, natürliche Auslese charakterisieren lassen. Dadurch wurden mehrere religiöse und weitere weltanschauliche Überzeugungen in Frage gestellt. Deshalb wird die Evolutionstheorie auch heute noch vielfach kritisiert, ja bekämpft. Die Vorwürfe lassen sich ordnen nach den Kriterien, mit denen wir erfahrungswissenschaftliche Theorien beurteilen. Haltbar ist daran nur, dass es für die Evolutionstheorie zwar beliebig viele Bestätigungen gibt, aber nur wenige Widerlegungsmöglichkeiten. Durch die neuerdings entwickelten und durchgeführten Evolutionsexperimente ist die empirische Situation allerdings deutlich besser geworden. Am (erfahrungs)wissenschaftlichen Charakter der Evolutionstheorie besteht deshalb kein Zweifel.

  3. Engineering the yeast Yarrowia lipolytica for the production of therapeutic proteins homogeneously glycosylated with Man8GlcNAc2 and Man5GlcNAc2

    PubMed Central

    2012-01-01

    Background Protein-based therapeutics represent the fastest growing class of compounds in the pharmaceutical industry. This has created an increasing demand for powerful expression systems. Yeast systems are widely used, convenient and cost-effective. Yarrowia lipolytica is a suitable host that is generally regarded as safe (GRAS). Yeasts, however, modify their glycoproteins with heterogeneous glycans containing mainly mannoses, which complicates downstream processing and often interferes with protein function in man. Our aim was to glyco-engineer Y. lipolytica to abolish the heterogeneous, yeast-specific glycosylation and to obtain homogeneous human high-mannose type glycosylation. Results We engineered Y. lipolytica to produce homogeneous human-type terminal-mannose glycosylated proteins, i.e. glycosylated with Man8GlcNAc2 or Man5GlcNAc2. First, we inactivated the yeast-specific Golgi α-1,6-mannosyltransferases YlOch1p and YlMnn9p; the former inactivation yielded a strain producing homogeneous Man8GlcNAc2 glycoproteins. We tested this strain by expressing glucocerebrosidase and found that the hypermannosylation-related heterogeneity was eliminated. Furthermore, detailed analysis of N-glycans showed that YlOch1p and YlMnn9p, despite some initial uncertainty about their function, are most likely the α-1,6-mannosyltransferases responsible for the addition of the first and second mannose residue, respectively, to the glycan backbone. Second, introduction of an ER-retained α-1,2-mannosidase yielded a strain producing proteins homogeneously glycosylated with Man5GlcNAc2. The use of the endogenous LIP2pre signal sequence and codon optimization greatly improved the efficiency of this enzyme. Conclusions We generated a Y. lipolytica expression platform for the production of heterologous glycoproteins that are homogenously glycosylated with either Man8GlcNAc2 or Man5GlcNAc2 N-glycans. This platform expands the utility of Y. lipolytica as a heterologous expression host

  4. TÜV - Zertifizierungen in der Life Science Branche

    NASA Astrophysics Data System (ADS)

    Schaff, Peter; Gerbl-Rieger, Susanne; Kloth, Sabine; Schübel, Christian; Daxenberger, Andreas; Engler, Claus

    Life Sciences [1] (Lebenswissenschaften) sind ein globales Innovationsfeld mit Anwendungen der Bio- und Medizinwissenschaften, der Pharma-, Chemie-, Kosmetik- und Lebensmittelindustrie. Diese Branche zeichnet sich durch eine stark interdisziplinäre Ausrichtung aus, mit Anwendung wissenschaftlicher Erkenntnisse und Einsatz von Ausgangsstoffen aus der modernen Biologie, Chemie und Humanmedizin sowie gezielter marktwirtschaftlich orientierter Arbeit.

  5. Historisches Rätsel Ein Pionier der Raumfahrt

    NASA Astrophysics Data System (ADS)

    Loos, Andreas

    2004-05-01

    Geschichten über seine Person muss man in der Regel mit Vorsicht genießen. Schließlich war der gesuchte Raketenforscher ein begabter Selbstdarsteller, der schon seine ersten größeren Versuche zu öffentlichen Flugschauen machte - und dafür Eintrittsgeld nahm.

  6. Role of surface-inoculated Debaryomyces hansenii and Yarrowia lipolytica strains in dried fermented sausage manufacture. Part 1: Evaluation of their effects on microbial evolution, lipolytic and proteolytic patterns.

    PubMed

    Patrignani, Francesca; Iucci, Luciana; Vallicelli, Melania; Guerzoni, M Elisabetta; Gardini, Fausto; Lanciotti, Rosalba

    2007-04-01

    The aim of this work was to study the effects of Debaryomyces hansenii and Yarrowia lipolytica strains, used with lactic acid starter cultures (Lactobacillus plantarum), on the manufacture of dried fermented sausages to understand their role on sausage microbial evolution, lipolytic and proteolytic patterns. The inoculation of the yeast strains did not markedly affect the sausage's microbial flora. The sausages with the yeast strains showed more marked and earlier water activity (a(w)) reductions. Moreover, the surface inoculation of the yeast strains resulted, at the end of ripening, in more pronounced proteolysis and lipolysis. The lipolytic patterns of the products were affected not only by the yeast strain but also by the level of mincing of the meat mixture used. PMID:22064033

  7. Evaluation of heterologous α-amylase production in two expression platforms dedicated for Yarrowia lipolytica: commercial Po1g-pYLSC (php4d) and custom-made A18-pYLTEF (pTEF).

    PubMed

    Celińska, Ewelina; Borkowska, Monika; Białas, Wojciech

    2016-05-01

    In view of the constantly increasing demand for cost-effective, low-energy and environmentally friendly industrial processes and household care products, enzyme production occupies an essential place in the field of biotechnology. Along with increasing demand for industrial and household care enzymes, the demand for heterologous expression platforms has also increased. Apart from the conventional hosts, e.g. Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris, routinely used in heterologous protein expression, the non-conventional ones have become more and more exploited in this field. Among the available yeast host systems, Yarrowia lipolytica appears to be an attractive alternative. The aim of this study was to compare efficiency of two Yarrowia-based expression platforms, commercial Po1g-pYLSC and custom-made A18-pYLTEF, in expression of an insect-derived, raw-starch-digesting α-amylase, to select the 'champion' system for further studies on this valuable enzyme. Both expression platforms were compared with respect to copy number of the integrated expression cassette/transformed genome, and the recombinant strains performance (Po1g-pYLSC-derived 4.29 strain, and A18-pYLTEF-derived B9 strain) during batch bioreactor cultures. Our results demonstrate that the average number of integration events into the recipient's genome was comparable for both expression systems under investigation, but with varying distribution of the multicopy integrants; and the number of the recombinant gene copies was highly correlated with the acquired amylolytic activity of the strains. Due to severe susceptibility of the recombinant AMY1 polypeptide to native proteases of the custom-made expression system, the final yield of the enzyme was substantially lower when compared to the commercial Po1g-pYLSC (reaching a maximum level of 142.84 AU/l). Copyright © 2015 John Wiley & Sons, Ltd. PMID:26694961

  8. Darwin, Engels und die Rolle der Arbeit in der biologischen und kulturellen Evolution des Menschen

    NASA Astrophysics Data System (ADS)

    Reichholf, Josef H.

    Im Jahre 1876, 5 Jahre nach Erscheinen von Darwins Buch über die Evolution des Menschen und die sexuelle Selektion (Darwin 1871), veröffentlichte Friedrich Engels den berühmt gewordenen Essay "Anteil der Arbeit an der Menschwerdung des Affen“ (Engels 1876). Die Kernfrage darin lautet in Kurzform: Warum hat der Mensch eigentlich ein Bedürfnis nach Arbeit? Engels Antwort wird nachfolgend näher betrachtet und vom gegenwärtigen Kenntnisstand aus beurteilt. Wie sich zeigen wird, beantworten seine Überlegungen die Frage nicht wirklich. Sie ist weiterhin offen. Es können lediglich einige zusätzliche Anhaltspunkte zur Diskussion gestellt werden. Angesichts des drängenden Problems millionenfacher Arbeitslosigkeit und der Forderungen nach einem "Grundrecht auf Arbeit“ kommt den Überlegungen zum möglichen Ursprung des Bedürfnisses nach Arbeit mehr als nur akademisches Interesse zu.

  9. Die Evolution der Religiosität

    NASA Astrophysics Data System (ADS)

    Voland, Eckart

    Ein konsequent darwinischer Blick auf den Menschen bedeutet, auch im Denken, Fühlen und Handeln biologische Anpassungsgeschichte zu suchen, denn auch die psychischen und mentalen Eigenheiten des Homo sapiens unterliegen der natürlichen Selektion. Lässt sich die religiöse Lebenspraxis von Menschen daher auch aus einer Fitnessperspektive betrachten?

  10. The Forced van der Pol Equation

    ERIC Educational Resources Information Center

    Fay, Temple H.

    2009-01-01

    We report on a study of the forced van der Pol equation x + [epsilon](x[superscript 2] - 1)x + x = F cos[omega]t, by solving numerically the differential equation for a variety of values of the parameters [epsilon], F and [omega]. In doing so, many striking and interesting trajectories can be discovered and phenomena such as frequency entrainment,…

  11. Obituary for Jan van der Pers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    After a short but valiant struggle against cancer, Jan van der Pers died on 29 April, 2006 in the hospital in Hilversum, The Netherlands, close to his home. Our conversations with Jan during the last months of his life showed the remarkable strength and positive attitude typical of him. Discussions...

  12. Auf der Suche nach extrasolaren Transitplaneten

    NASA Astrophysics Data System (ADS)

    Heller, René

    2010-06-01

    Planeten um andere Sonnen, die von der Erde aus gesehen einmal während ihres Orbits vor ihrem Zentralstern vorbeiziehen, eröffnen eine bis vor Kurzem ungeahnte Palette an Möglichkeiten zu ihrer Untersuchung. Nur: Wo am Himmel lassen sich diese Kandidaten für Sternbedeckungen eigentlich finden?

  13. Chirurgische Behandlung von Melanomen in der Schwangerschaft: eine praktische Anleitung.

    PubMed

    Crisan, Diana; Treiber, Nicolai; Kull, Thomas; Widschwendter, Peter; Adolph, Oliver; Schneider, Lars Alexander

    2016-06-01

    Als ein Tumor, der primär eine chirurgische Behandlung erfordert, ist ein neu diagnostiziertes oder vorbestehendes Melanom in der Schwangerschaft eine klinische Rarität. In solchen Fällen steht der Chirurg vor der Herausforderung, ein geeignetes therapeutisches Vorgehen festlegen zu müssen. Auf der Grundlage unserer klinischen Erfahrung und einer Übersicht über die Literatur geben wir in der vorliegenden Arbeit eine Anleitung für das praktische Vorgehen bei dieser seltenen klinischen Konstellation. Unserer Erfahrung nach müssen schwangere Melanom-Patientinnen im Hinblick auf ihre therapeutischen Optionen ausführlich beraten werden. Naturgemäß setzen sie ihr ungeborenes Kind an die erste Stelle und zögern, der erforderlichen Operation zuzustimmen, obwohl bei ihnen eine möglicherweise lebensbedrohliche Erkrankung diagnostiziert worden ist. Daher ist es entscheidend, diese Patientinnen klar darüber zu informieren, dass, wie die vorliegenden medizinischen Erfahrungen zeigen, eine Schwangerschaft per se kein Grund ist, eine notwendige Melanom-Operation aufzuschieben. Jedoch müssen bei einigen Parametern wie den präoperativen Bildgebungsverfahren, der Positionierung auf dem Operationstisch, der Überwachung, Anästhesie und der perioperativen Medikation bestimmte Anpassungen vorgenommen werden, um der speziellen Situation Rechnung zu tragen. PMID:27240063

  14. Die neogene Hebungsgeschichte der Patagonischen Anden im Kontext der Subduktion eines aktiven Spreizungszentrums

    NASA Astrophysics Data System (ADS)

    Warkus, Frank

    2002-05-01

    Das Phänomen der Subduktion eines aktiven Spreizungszentrums an der Südspitze Südamerikas ist seit langem bekannt. Eine Vielzahl von geologischen Beobachtungen wurden mit diesem Phänomen in Verbindung gebracht, trotzdem ist der genaue Mechanismus der Beeinflussung des aktiven Kontinentalrandes weitgehend unbekannt. Die Zusammenhänge zwischen den Subduktionsprozessen und der Entwicklung der patagonischen Anden zwischen 47°S und 48°S stehen im Mittelpunkt der Untersuchungen. Um eine detaillierte zeitliche Auflösung der zugrunde liegenden Prozesse untersuchen zu können, wurde die Entwicklung der Vorlandsedimentation, die thermische Entwicklung und die Heraushebung der Oberkruste des andinen Orogens untersucht und diese in Bezug zur Subduktion des Chile-Rückens gesetzt. Im Bereich von 47°30′S wurden die synorogenen Vorlandsedimente der Santa Cruz Formation sedimentologisch untersucht. Diese fluviatilen Sedimente wurden in einem reliefarmen Vorlandgebiet durch häufige Rinnenverlagerung und dem Aufbau von Rinnenumlagerungsgürteln in Kombination mit assoziierten groräumigen Überflutungsablagerungen akkumuliert. Sie stehen in einem engen Zusammenhang mit der orogenen Entwicklung im andinen Liefergebiet. Dies spiegelt sich in dem nach oben gröber werdenden Zyklus der Santa Cruz Formation wider. Die magnetostratigraphischen Untersuchungen einer 270 m mächtigen Sequenz aus der Basis der Santa Cruz Formation, die mit 329 Einzelproben aus 96 Probenpunkten beprobt wurde, ergab 7 Umkehrungen der geomagnetischen Feldrichtung. Mit Hilfe der geomagnetischen Polaritätszeitskala (CANDE AND KENT, 1995) konnte der untersuchte Abschnitt der Santa Cruz Formation zwischen 16.2 und 18.5 Ma datiert werden. Als Träger der Sedimentations-Remanenz konnten überwiegend Pseudoeinbereichs-Magentitpartikel und untergeordnet Hämatitpartikel identifiziert werden. An drei Profilen der Santa Cruz Formation wurden aus Sandsteinlagen unterschiedlicher stratigraphischer Position

  15. Über die Methode der physikalischen Naturbeschreibung [33

    NASA Astrophysics Data System (ADS)

    Wenn es zu den Aufgaben der Philosophie und der wissensehaftlicher Theologie gehört, das Warum und Wie unserer Existenz und ihrer Umwelt zu ergründen, so muss als ihr Ausgangspunkt eine vollständige und möglichst übersichtliche Beschreibung unserer Empfindungen vorliegen. Einen Teil dieser Empfindungen fassen wir unter dem Namen Sinnesempfindungen zusammen. Ob ein solcher Aussehnitt genau abgegrenzt werden kann, soll hier nicht untersucht werden. Es kann sogar mit gewisser Berechtigung behauptet werden, dass eine Unterteilung gar nicht streng durchführbar ist. Der schon in der Philosophie der Antike uns entgegentretende Gedanke, dass die Welt nur in ihrer Gesamtheit rerständlich sein kann, kommt sicherlich der Wahrheit näher als die philosophischen Systeme des verflossenen Jahrhunderts, welche glaubten, dass alies aus unseren materialistischen Erkenntnissen heraus erklärt werden könne. Die Entstehung dieser Systeme, ich denke z. Â. an den Monisimis Haeckels, erklärt sich aus der Hybris, welche die Reaktion der Philosophie auf die bedeutenden Erfolge der exakten Naturwissenschaften war. Die ebenfalls nicht zu imterschätzenden Fortschritte unserer heutigen Kenntnisse auf diesem Gebiete zwingen din Naturforschung eher zu einer immer steigenden Bescheidenheit den "Welträtseln" gegenüber, deren letzte Konsequenz vielleicht darin liegen wird, dass sie die eingangs der Philosophie und Theologie zugewiesene Aufgabe als unlosbar bezeichnen muss..

  16. Resonance broadening and van der waals broadening

    NASA Astrophysics Data System (ADS)

    Mashonkina, L.

    2010-11-01

    Resonance broadening is important for the hydrogen lines in the spectra of F-type and later stars. In the corresponding temperature regime, the extended wings of the Balmer lines are used as a stellar effective temperature indicator. We show the effect of the use of two broadening theories, Ali & Griem (1965, 1966) and Barklem et al. (2000a, 2000b), on the effective temperature derived in non-LTE from Hα and Hβ in the Sun and the metal-poor star HD19445. Van der Waals broadening is important for strong spectral lines in the atmospheres of F-type and later stars. For the selected transitions in Ca I and Ca II, line profile comparisons are made between applying the van der Waals damping constants from laboratory measurements, the ABO perturbation theory, and the classic Unsöld approximation.

  17. van der Waals Forces between Cylinders

    PubMed Central

    Mitchell, D. J.; Ninham, B. W.; Richmond, P.

    1973-01-01

    We derive the retarded van der Waals interaction between two long thin parallel dielectric cylinders immersed in a solvent. The result shows that the ultraviolet interactions which may be responsible for the specificity of macromolecular interactions do not operate strongly over distances R ≥ 50 Å. For greater distances the contribution of these frequencies ξ is damped by a factor ∞ e-ξR/c. PMID:4696763

  18. Mehr Mathematik Wagen in der Medizin

    NASA Astrophysics Data System (ADS)

    Deuflhard, Peter; Dössel, Olaf; Louis, Alfred K.; Zachow, Stefan

    In diesem Artikel wird an drei Erfolgsmodellen dargestellt, wie das Zusammenwirken von Mathematik und Medizin eine Entwicklung hin zu patientenspezifischen Modellen auf Basis moderner medizinischer Bildgebung angestoßen hat, die in naher Zukunft dynamisch weiter Raum greifen wird. Dabei existiert ein Gleichklang der Interessen von Medizin und Mathematik: Beide Disziplinen wollen die Resultate schnell und zuverlässig. Für die Klinik heißt dies, dass notwendige Rechnungen in möglichst kurzer Zeit, und zwar auf dem PC, ablaufen müssen und dass die Resultate so genau und belastbar sein müssen, dass medizinische Entscheidungen darauf aufbauen können. Für die Mathematik folgt daraus, dass höchste Anforderungen an die Effizienz der verwendeten Algorithmen und die darauf aufbauende Software in Numerik und Visualisierung zu stellen sind. Allerdings ist es noch ein weiter Weg, bis anatomische und medizinisch brauchbare funktionelle Modelle auch nur für die wichtigsten Körperteile und die häufigsten Krankheitsfälle verfügbar sein werden. Führende Universitätskliniken könnten, als Zentren einer interdisziplinären Kooperation von Medizinern, Ingenieuren und Mathematikern, eine Vorreiterrolle dabei übernehmen, mehr Mathematik in der Medizin zu wagen. Dies wäre zweifellos ein wichtiger Schritt in Richtung auf eine individuelle quantitative Medizin, bei dem Deutschland die besten Voraussetzungen hätte, die Rolle des "Schrittmachers“ zu übernehmen.

  19. Zellulare Nichtlineare Netzwerke: Anwendungen in der Informationstechnologie

    NASA Astrophysics Data System (ADS)

    Tetzlaff, R.

    2007-06-01

    Zellulare Nichtlineare Netzwerke (CNN) wurden 1988 von Chua und Yang (Chua und Yang, 1988) eingeführt. Diese Netzwerke sind dadurch gekennzeichnet, dass eine Zelle, die die kleinste Einheit eines CNN darstellt, nur mit Zellen innerhalb einer bestimmten Umgebung verbunden ist. üblicherweise sind Art und Stärke der Wechselwirkung zwischen zwei Zellen eines CNN translationsinvariant, d.h. sie hängen nur von der relativen Lage beider Zellen zueinander ab. Im Vordergrund aktueller Arbeiten stehen auf derartigen Netzwerken basierende schaltungstechnische Realisierungen mit bis zu 176x144 Zellen, die eine direkte Verbindung zu zweidimensionalen optischen Sensor-Anordnungen aufweisen. Über einen separaten Speicherbereich können die Zellkopplungen eines Netzwerks verändert werden, wodurch eine adaptive Verarbeitung von mehrdimensionalen Sensorsignalen ermöglicht wird. Das kürzlich vorgestellte so genannte EyeRis System (Anafocus Ltd.) enthält zusätzlich noch einen Standardprozessor und stellt (bei einer Größe vergleichbar mit der einer Kreditkarte) daher ein vollständiges superschnelles System zur Informationsverarbeitung dar. In diesem Beitrag sollen, nach einem kurzen Überblick über die Eigenschaften von CNN, aktuelle Realisierungen und exemplarisch eine neuere eigene Anwendung vorgestellt und besprochen werden.

  20. Zum Wissenschaftsverständnis der modernen Evolutionsbiologie

    NASA Astrophysics Data System (ADS)

    Sommer, Ralf J.

    Die moderne Evolutionsbiologie hat ihren Ursprung in den Arbeiten von Charles Darwin und Alfred Wallace (Darwin 1963). Der gemeinsame Ausgangspunkt des Evolutionsgedanken ist dabei die Beobachtung, dass die biologische Welt nicht konstant ist. Biologische Systeme und alle darin lebenden Organismen unterliegen über längere Zeiträume hinweg einer stetigen Veränderung. Diese grundlegende Eigenschaft biologischer Systeme macht die Biologie zu einer historischen Wissenschaft und stellt einen wichtigen Gegensatz zu großen Teilen der Physik dar. Obwohl die Aussage von der Veränderlichkeit der Arten heute trivial klingt, war sie im 19. Jahrhundert eine Revolution, da die Konstanz der Arten und der Welt eine vorherrschende Stellung im damaligen Weltbild hatte (Amundson 2005).

  1. Der Telemanipulator daVinci als mechanisches Trackingsystem

    NASA Astrophysics Data System (ADS)

    Käst, Johannes; Neuhaus, Jochen; Nickel, Felix; Kenngott, Hannes; Engel, Markus; Short, Elaine; Reiter, Michael; Meinzer, Hans-Peter; Maier-Hein, Lena

    Der Telemanipulator daVinci (Intuitive Surgical, Sunnyvale, Kalifornien) ist ein M aster-Slave System für roboterassistierte minimalinvasive Chirurgie. Da er über integrierte Gelenksensoren verfügt, kann er unter Verwendung der daVinci-API als mechanisches Trackingsystem verwendet werden. In dieser Arbeit evaluieren wir die Präzision und Genauigkeit eines daVinci mit Hilfe eines Genauigkeitsphantoms mit bekannten Maßen. Der ermittelte Positionierungsfehler liegt in der Größenordnung von 6 mm und ist somit für einen Großteil der medizinischen Fragestellungen zu hoch. Zur Reduktion des Fehlers schlagen wir daher eine Kalibrierung der Gelenksensoren vor.

  2. Jahre Entwicklung der Instandhaltung - von der ausfallorientierten Instandhaltung zum gemeinsamen TPM und RCM

    NASA Astrophysics Data System (ADS)

    Iske, Friedhelm

    Zur Einleitung meines Beitrages möchte ich von einem Gespräch mit einem Mitarbeiter berichten, das ich als junger Vorgesetzter einer Instandhaltungsgruppe 1988 führte. Der engagierte Mitarbeiter feierte damals sein vierzigjähriges Dienstjubiläum und war stolz auf das von ihm Geleistete sowie auf den besonderen Einsatz seiner Altersgruppe, die nach dem Zweiten Weltkrieg das Werk wieder aufgebaut hatte. Auf meine Frage, was denn damals die erste Aufgabe in der Firma war, bekam ich kurz und knapp und mit einer Selbstverständlichkeit die selbstbewusste Antwort: "Unser Pferd füttern und mit dem Pferd die innerbetrieblichen Transporte erledigen“. Als junger, technisch orientierter Vorgesetzter war ich über diese Antwort sehr überrascht. Gedanklich weit entfernt war die Vorstellung, dass in der Vergangenheit Transporte mit einem Pferd erledigt wurden.

  3. Bereits nach Ablauf der Halbwertszeit droht der vollständige Zerfall Die britische Atomic Scientists’ Association, die Ideologie der „objektiven” Wissenschaft und die H-Bombe

    NASA Astrophysics Data System (ADS)

    Laucht, Christoph

    Präsident Harry Trumans Verlautbarung vom 31.1.1950, seine Regierung wolle die Entwicklung der Wasserstoffbombe vorantreiben, fand große Beachtung in den britischen Medien. Die illustrierte Zeitschrift Picture Post widmete der HBombe einen Artikel, der unter anderem kurze Stellungnahmen der britischen Atomwissenschaftler Eric Burhop, Kathleen Lonsdale, Harrie Massey, Rudolf Peierls und Maurice Pryce enthielt, die alle Mitglieder der Atomic Scientists' Association (ASA) waren.

  4. Information Interaction Study for DER and DMS Interoperability

    NASA Astrophysics Data System (ADS)

    Liu, Haitao; Lu, Yiming; Lv, Guangxian; Liu, Peng; Chen, Yu; Zhang, Xinhui

    The Common Information Model (CIM) is an abstract data model that can be used to represent the major objects in Distribution Management System (DMS) applications. Because the Common Information Model (CIM) doesn't modeling the Distributed Energy Resources (DERs), it can't meet the requirements of DER operation and management for Distribution Management System (DMS) advanced applications. Modeling of DER were studied based on a system point of view, the article initially proposed a CIM extended information model. By analysis the basic structure of the message interaction between DMS and DER, a bidirectional messaging mapping method based on data exchange was proposed.

  5. Phänomenologische Grundlagen der Wärmelehre

    NASA Astrophysics Data System (ADS)

    Heintze, Joachim

    Die Physik der Wärme lässt sich auf zweierlei Weise formulieren: Einmal als Mechanik eines Systems, das eine enorm große Zahl von Teilchen enthält (statistische Mechanik), und einmal mit Hilfe von ad hoc eingeführten Größen, den sogenannten Zustandsgrößen, die geeignet sind, das Verhalten eines solchen Systems zu beschreiben, ohne dass man die Teilchen selbst und ihre Bewegungen betrachten muss (Thermodynamik). Wir werden beide Ansätze in ihrer einfachsten und anschaulichsten Ausprägung in Kap. 5 (kinetische Gastheorie) bzw. in Kap. 8 (Grundbegriffe der Thermodynamik) behandeln. Obgleich sich die volle Durchführung des Programms als begrifflich und mathematisch recht schwierig erweist - die Vorlesung "Thermodynamik und Statistik" steht gewöhnlich am Ende der Kursvorlesungen über theoretische Physik - werden wir doch auf der Grundlage der Kap. 5 und 8 eine Menge über die Physik der Wärme lernen können. Den Ausgangspunkt der Wärmelehre bilden jedoch allemal die Naturerscheinungen, die wir hier in Kap. 4 behandeln wollen: Wärme, Kälte, Temperaturausgleich. Wir werden untersuchen, wie man diese Begriffe quantifizieren kann, und was bei der Erwärmung oder Abkühlung eines Körpers vor sich geht. Dabei werden wir auf den I. und II. Hauptsatz der Wärmelehre stoßen.

  6. Biological activity of recombinant Der p 2, Der p 5 and Der p 7 allergens of the house-dust mite Dermatophagoides pteronyssinus.

    PubMed

    Lynch, N R; Thomas, W R; Garcia, N M; Di Prisco, M C; Puccio, F A; L'opez, R I; Hazell, L A; Shen, H D; Lin, K L; Chua, K Y

    1997-09-01

    Der p 2, Der p 5 and Der p 7 are three allergens of the house-dust mite Dermatophagoides pteronyssinus that have been cloned and expressed in Escherichia coli as fusion proteins with glutathione-S-transferase (GST). We showed that these recombinant allergens produced immediate hypersensitivity skin-test reactions in 70, 60 and 52% respectively of a group of mite-sensitive allergic patients who were strongly positive to whole mite extract (WME). Comparable positivities were found for serum levels of specific IgE antibody against these allergens, as measured by the radioallergosorbant test (RAST). Overall, for the group of allergic patients that we evaluated, the serum IgE antibody concentrations against Der p 2, 5 and 7 were calculated to represent about one third, one quarter and one fifth respectively of the levels measured against the WME. However, for some patients the activity determined against the separate allergens was far higher than that detected against the WME, thus indicating that the concentration of these can be limiting in the WME. We found no significant correlations between the RAST levels against Derp 2 and either Derp 5 or 7, and RAST-inhibition tests indicated a lack of cross-reactivity between Der p 2 and the other two allergens. In contrast, the RAST results revealed the existence of a significant immunological relationship between Der p 5 and 7. Although a certain degree of reactivity against the GST fusion partner was found in the allergic patients studied, this was not a significant influence in determining the positivity against the recombinant allergens. These results confirm the in vivo biological activity of recombinant Der p 2, 5 and 7, and indicate that whilst Der p 2 is undoubtedly a major mite allergen, both Der p 5 and 7 make important contributions toward the overall allergenic activity of house-dust mites. PMID:9303332

  7. Die Deutsche Statistische Gesellschaft in der Weimarer Republik und während der Nazidiktatur

    NASA Astrophysics Data System (ADS)

    Wilke, Jürgen

    Nach anfänglichen Schwierigkeiten durch den 1. Weltkrieg erlangte die Deutsche Statistische Gesellschaft (DStatG) unter dem renommierten Statistiker und Vorsitzenden der DStatG, Friedrich Zahn, durch eine Vielzahl von Aktivitäten hohes Ansehen. Es gab Bestrebungen, Statistiker aus allen Arbeitsfeldern der Statistik in die DStatG zu integrieren, wobei die "Mathematische Statistik" nur zögerlich akzeptiert wurde (Konjunkturforschung, Zeitreihenanalyse). Nach der Machtübernahme 1933 durch Adolf Hitler geriet die DStatG in das Fahrwasser nationalsozialistischer Ideologie und Politik (Führerprinzip, Gleichschaltung des Vereinswesens). Damit war eine personelle Umstrukturierung in der DStatG verbunden. Politisch Missliebige und rassisch Verfolgte mussten die DStatG verlassen (Bernstein, Freudenberg, Gumbel u.a.). Unter den Statistikern gab es alle Abstufungen im Verhalten zum Regime von Ablehnung und zwangsweiser Anpassung über bereitwilliges Mitläufertum bis zu bewusster Täterschaft. Besonders die Bevölkerungsstatistik wurde durch die NS- Rassenpolitik auf lange Sicht diskreditiert. Im Rahmen von Wirtschaftsplanung und Aufrüstung wurden neue zukunftsträchtige statistische Modelle (Grünig, Bramstedt, Leisse) entwickelt.

  8. van der Waals Heterostructures Grown by MBE

    NASA Astrophysics Data System (ADS)

    Hinkle, Christopher

    In this work, we demonstrate the high-quality MBE heterostructure growth of various layered 2D materials by van der Waals epitaxy (VDWE). The coupling of different types of van der Waals materials including transition metal dichalcogenide thin films (e.g., WSe2, WTe2, HfSe2) , insulating hexagonal boron nitride (h-BN), and topological insulators (e.g., Bi2Se3) allows for the fabrication of novel electronic devices that take advantage of unique quantum confinement and spin-based characteristics. The relaxed lattice-matching criteria of van der Waals epitaxy has allowed for high-quality heterostructure growth with atomically abrupt interfaces, allowing us to couple these materials based primarily on their band alignment and electronic properties. We will discuss the impact of sample preparation, surface reactivity, and lattice mismatch of various substrates (sapphire, graphene, TMDs, Bi2Se3) on the growth mode and quality of the films and will discuss our studies of substrate temperature and flux rates on the resultant growth and grain size. Structural and chemical characterization was conducted via reflection high energy electron diffraction (RHEED, X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning tunneling microscopy/spectroscopy (STM/S), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy. Experimentally determined band alignments have been determined and compared with first-principles calculations allowing the design of novel low-power logic and magnetic memory devices. Initial results from the electrical characterization of these grown thin films and some simple devices will also be presented. These VDWE grown layered 2D materials show significant potential for fabricating novel heterostructures with tunable band alignments and magnetic properties for a variety of nanoelectronic and optoelectronic applications.

  9. Kanban - der Weg ist das Ziel

    NASA Astrophysics Data System (ADS)

    Aull, Florian; Berlak, Joachim; Dickmann, Eva; Dickmann, Philipp; Fischäder, Holm; Gerlach, Joachim; Henneberg, Jens; Kapalla, Klaus; Kress, Oliver; Kuttler, Robert; Schneider, Herfried M.; Schürle, Philipp; Stellpflug, Franz-Josef; Wannenwetsch, Ralph; Wulz, Johannes; Zäh, Michael F.

    Wenn man aktuell Produktionsbereiche in Deutschland und Europa besucht, fallen im Zusammenhang mit modernen Produktionsmethoden immer öfter die Begriffe Kanban (jap. Karte, Signal) und Pull-Produktion, und dies nicht ohne Stolz, da diese mit dem schillernden Vorbild des Toyota Produktionssystems in Zusammenhang stehen. Tatsächlich ist Kanban ein integraler Bestandteil moderner Produktionssysteme. Blickt man aber im Rahmen von Prozessanalysen hinter die Fassaden", d. h. in die tägliche Praxis der Arbeitsprozesse, wird man schnell desillusioniert - die viel gepriesenen klassischen Kanban-Regeln werden im Tagesgeschäft nicht eingehalten.

  10. Komplexität der Geographie

    NASA Astrophysics Data System (ADS)

    Diekert, Volker; Hertrampf, Ulrich

    Das allgemein als Prototyp eines PSPACE-vollständigen Spiels gesehene Geographiespiel wird bezüglich seiner Komplexität genauer untersucht. Das Interesse der theoretischen Informatik an diesem Spiel wurde sehr durch die Darstellung in dem Lehrbuch von Papadimitriou [Pap94] gefördert. Allerdings bestimmt dieses Lehrbuch nicht die Komplexität des Standardspiels sondern verwendet eine Verallgemeinerung. Die Aussage in dem Lehrbuch bleibt damit etwas unbefriedigend und hinter den Möglichkeiten. Wir zeigen hier, dass die komplexitätstheoretische Charakterisierung schon für die Standardvariante des Spiels gilt.