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Anna Yurievna Arinbasarova, Andrey Valerievich Machulin, Elena Nikolaevna Biryukova, Vladimir Vladislavovich Sorokin, Alexander Grigorievich Medentsev, Nataliya Egorovna Suzina
The ultrastructural changes in the cell envelope of the yeast Yarrowia lipolytica as stress response were examined using electron microscopy technique. The formation of new cellular surface structures including membrane vesicles, pore channels and wall surface globules were shown for the first time under conditions of oxidative (endogenous and exogenous) or thermal stress. This demonstrates once again that under stress conditions the microorganisms reveal properties unknown for them before. Particularly noteworthy is the silicon accumulation, which was revealed at the surface globules with X-ray microanalysis of the elemental composition of cells' thin sections...
February 14, 2018: Canadian Journal of Microbiology
Sangwoo Kim, Kyung-Jin Kim
Acyl-CoA oxidases (ACOXs) play important roles in lipid metabolism, including peroxisomal fatty acid β-oxidation by the conversion of acyl-CoAs to 2-trans-enoyl-CoAs. The yeast Yarrowia lipolyticacan utilize fatty acidsas a carbon source and thus has extensive biotechnological applications. The crystal structure of ACOX3from Y. lipolytica(YlACOX3) was determinedat a resolution of2.5 Å. It contained two molecules per asymmetric unit, and the monomeric structure was folded into four domains, Nα, Nβ, Cα1 and Cα2domains...
February 13, 2018: Journal of Microbiology and Biotechnology
Xochitl Niehus, Anne-Marie Crutz-Le Coq, Georgina Sandoval, Jean-Marc Nicaud, Rodrigo Ledesma-Amaro
Background: Yarrowia lipolytica is a common biotechnological chassis for the production of lipids, which are the preferred feedstock for the production of fuels and chemicals. To reduce the cost of microbial lipid production, inexpensive carbon sources must be used, such as lignocellulosic hydrolysates. Unfortunately, lignocellulosic materials often contain toxic compounds and a large amount of xylose, which cannot be used by Y. lipolytica. Results: In this work, we engineered this yeast to efficiently use xylose as a carbon source for the production of lipids by overexpressing native genes...
2018: Biotechnology for Biofuels
Si-Jia Xue, Zhe Chi, Yu Zhang, Yan-Feng Li, Guang-Lei Liu, Hong Jiang, Zhong Hu, Zhen-Ming Chi
PURPOSE: Oleaginous yeasts, fatty acids biosynthesis and regulation in the oleaginous yeasts and the fatty acids from the oleaginous yeasts and their applications are reviewed in this article. RESULTS: Oleaginous yeasts such as Rhodosporidium toruloides, Yarrowia lipolytica, Rhodotorula mucilaginosa, and Aureobasidium melanogenum, which can accumulate over 50% lipid of their cell dry weight, have many advantages over other oleaginous microorganisms. The fatty acids from the oleaginous yeasts have many potential applications...
February 1, 2018: Critical Reviews in Biotechnology
Matthew Brabender, Murtaza Shabbir Hussain, Gabriel Rodriguez, Mark A Blenner
Yarrowia lipolytica is an industrial yeast that has been used in the sustainable production of fatty acid-derived and lipid compounds due to its high growth capacity, genetic tractability, and oleaginous properties. This investigation examines the possibility of utilizing urea or urine as an alternative to ammonium sulfate as a nitrogen source to culture Y. lipolytica. The use of a stoichiometrically equivalent concentration of urea in lieu of ammonium sulfate significantly increased cell growth when glucose was used as the carbon source...
January 30, 2018: Applied Microbiology and Biotechnology
Carina Holkenbrink, Marie Inger Dam, Kanchana Rueksomtawin Kildegaard, Johannes Beder, Jonathan Dahlin, David Doménech Belda, Irina Borodina
The oleaginous yeast Yarrowia lipolytica is an emerging host for production of fatty acid-derived chemicals. To enable rapid iterative metabolic engineering of this yeast, there is a need for well-characterized genetic parts and convenient and reliable methods for their incorporation into yeast. Here, we present the EasyCloneYALI genetic toolbox, which allows streamlined strain construction with high genome editing efficiencies in Y. lipolytica via the CRISPR/Cas9 technology. The toolbox allows marker-free integration of gene expression vectors into characterized genome sites as well as marker-free deletion of genes with the help of CRISPR/Cas9...
January 29, 2018: Biotechnology Journal
Antônio Jesus Dorighetto Cogo, Keilla Dos Reis Dutra Ferreira, Lev A Okorokov, Alessandro C Ramos, Arnoldo R Façanha, Anna L Okorokova-Façanha
Polyamines play a regulatory role in eukaryotic cell growth and morphogenesis. Despite many molecular advances, the underlying mechanism of action remains unclear. Here, we investigate a mechanism by which spermine affects the morphogenesis of a dimorphic fungal model of emerging relevance in plant interactions, Yarrowia lipolytica, through the recruitment of a phytohormone-like pathway involving an activation of the plasma membrane P-type H+-ATPase. Morphological transition was followed microscopically and the H+-ATPase activity was analyzed in isolated membrane vesicles...
January 22, 2018: Biology Open
Hamid Mukhtar, Syed M Suliman, Aroosh Shabbir, Muhammad W Mumtaz, Umer Rashid, Sawsan A Rahimuddin
Fifty yeast strains were isolated from soil and eleven of them were selected for oil production. Five yeast strains were designated as oleaginous because they produced more than 16% of oil based on their biomass. The isolate IIB-10 identified as Yarrowia lipolytica produced maximum amount of lipids i.e. 22.8%. More amount of biomass was obtained when cane molasses was utilized as carbon source where it produced 29.4 g/L of biomass while sucrose and lactose were not utilized by IIB-10 and no biomass was obtained...
January 21, 2018: Protein and Peptide Letters
Jinyong Yan, Bingnan Han, Xiaohua Gui, Guilong Wang, Li Xu, Yunjun Yan, Catherine Madzak, Dujie Pan, Yaofeng Wang, Genhan Zha, Liangcheng Jiao
Lipases are scarcely exploited as feed enzymes in hydrolysis of lipids for increasing energy supply and improving nutrient use efficiency. In this work, we performed homologous overexpression, in vitro characterization and in vivo assessment of a lipase from the yeast Yarrowia lipolytica for feed purpose. Simultaneously, a large amount of yeast cell biomass was produced, for use as single cell protein, a potential protein-rich feed resource. Three kinds of low cost agro-industrial wastes were tested as substrates for simultaneous production of lipase and single cell protein (SCP) as feed additives: sugarcane molasses, waste cooking oil and crude glycerol from biodiesel production...
January 15, 2018: Scientific Reports
Allison Yaguchi, Michael Spagnuolo, Mark Blenner
Realizing the economic benefits of alternative substrates for commodity chemical bioproduction typically requires significant metabolic engineering of common model organisms, such as Saccharomyces cerevisiae. A growing toolkit is enabling engineering of non-conventional yeast that have robust native metabolism for xylose, acetate, aromatics, and waste lipids. Scheffersomyces stipitis was engineered to produce itaconic acid from xylose. Yarrowia lipolytica produced lipids from dilute acetate at over 100g/L. Cutaneotrichosporon oleaginosus was engineered to produce omega-3 fatty acids and recently was shown to accumulate nearly 70% lipids when grown on aromatics as a carbon source...
January 10, 2018: Current Opinion in Biotechnology
Hana Raschmanova, Astrid Weninger, Anton Glieder, Karin Kovar, Thomas Vogl
Within five years, the CRISPR-Cas system has emerged as the dominating tool for genome engineering, while also changing the speed and efficiency of metabolic engineering in conventional (Saccharomyces cerevisiae and Schizosaccharomyces pombe) and non-conventional (Yarrowia lipolytica, Pichia pastoris syn. Komagataella phaffii, Kluyveromyces lactis, Candida albicans and C. glabrata) yeasts. Especially in S. cerevisiae, an extensive toolbox of advanced CRISPR-related applications has been established, including crisprTFs and gene drives...
January 10, 2018: Biotechnology Advances
Ann-Kathrin Löbs, Cory Schwartz, Ian Wheeldon
Microbial production of chemicals and proteins from biomass-derived and waste sugar streams is a rapidly growing area of research and development. While the model yeast Saccharomyces cerevisiae is an excellent host for the conversion of glucose to ethanol, production of other chemicals from alternative substrates often requires extensive strain engineering. To avoid complex and intensive engineering of S. cerevisiae, other yeasts are often selected as hosts for bioprocessing based on their natural capacity to produce a desired product: for example, the efficient production and secretion of proteins, lipids, and primary metabolites that have value as commodity chemicals...
September 2017: Synthetic and Systems Biotechnology
Carlos Eduardo Robles-Rodríguez, Rafael Muñoz-Tamayo, Carine Bideaux, Nathalie Gorret, Stéphane E Guillouet, Carole Molina-Jouve, Gilles Roux, César Arturo Aceves-Lara
Oleaginous yeasts have been seen as a feasible alternative to produce the precursors of biodiesel due to their capacity to accumulate lipids as triacylglycerol having profiles with high content of unsaturated fatty acids. The yeast Yarrowia lipolytica is a promising microorganism that can produce lipids under nitrogen depletion conditions and excess of the carbon source. However, under these conditions, this yeast also produces citric acid (overflow metabolism) decreasing lipid productivity. This work presents two mathematical models for lipid production by Yarrowia lipolytica from glucose...
December 30, 2017: Biotechnology and Bioengineering
Cory Schwartz, Keith Frogue, Joshua Misa, Ian Wheeldon
Carotenoids are a class of molecules with commercial value as food and feed additives with nutraceutical properties. Shifting carotenoid synthesis from petrochemical-based precursors to bioproduction from sugars and other biorenewable carbon sources promises to improve process sustainability and economics. In this work, we engineered the oleaginous yeast Yarrowia lipolytica to produce the carotenoid lycopene. To enhance lycopene production, we tested a series of strategies to modify host cell physiology and metabolism, the most successful of which were mevalonate pathway overexpression and alleviating auxotrophies previously engineered into the PO1f strain of Y...
2017: Frontiers in Microbiology
Zhong-Peng Guo, Sophie Duquesne, Sophie Bozonnet, Jean-Marc Nicaud, Alain Marty, Michael Joseph O'Donohue
Background: A recently constructed cellulolytic Yarrowia lipolytica is able to grow efficiently on an industrial organosolv cellulose pulp, but shows limited ability to degrade crystalline cellulose. In this work, we have further engineered this strain, adding accessory proteins xylanase II (XYNII), lytic polysaccharide monooxygenase (LPMO), and swollenin (SWO) from Trichoderma reesei in order to enhance the degradation of recalcitrant substrate. Results: The production of EG I was enhanced using a promoter engineering strategy...
2017: Biotechnology for Biofuels
Sven Bordewick, Andy Beier, Kathleen Balke, Uwe T Bornscheuer
Nine new putative Baeyer-Villiger monooxygenase encoding genes were identified in the eukaryote Yarrowia lipolytica and eight were subsequently cloned and expressed. These enzymes, Yarrowia monooxygenases A-H (YMOA-H), were used in biocatalysis reactions with ketones, sulfides and sulfoxides as substrates. YMOB converts ketones and sulfides, albeit with low activities. However, YMOA did not convert any of the tested ketone substrates, but showed activity towards sulfides and sulfoxides and also showed very high stereoselectivity...
February 2018: Enzyme and Microbial Technology
Sangwoo Kim, Kyung-Jin Kim
Acyl-CoA oxidase (ACOX) plays an important role in fatty acid degradation. The enzyme catalyzes the first reaction in peroxisomal fatty acid β-oxidation by reducing acyl-CoA to 2-trans-enoyl-CoA. The yeast Yarrowia lipolytica is able to utilize fatty acids, fats, and oil as carbon sources to produce valuable bioproducts. We determined the crystal structure of ACOX1 from Y. lipolytica (YlACOX1) at a resolution of 2.5 Å. YlACOX1 forms a homodimer, and the monomeric structure is composed of four domains, the Nα, Nβ, Cα1, and Cα2...
November 30, 2017: Biochemical and Biophysical Research Communications
Ja Kyong Ko, Sun-Mi Lee
Cellulosic fuels are expected to have great potential industrial applications in the near future, but they still face technical challenges to become cost-competitive fuels, thus presenting many opportunities for improvement. The economical production of viable biofuels requires metabolic engineering of microbial platforms to convert cellulosic biomass into biofuels with high titers and yields. Fortunately, integrating traditional and novel engineering strategies with advanced engineering toolboxes has allowed the development of more robust microbial platforms, thus expanding substrate ranges...
November 27, 2017: Current Opinion in Biotechnology
Klára Herkommerová, Jana Zemančíková, Hana Sychrová, Zuzana Antošová
OBJECTIVES: To improve the storage stability and reusability of various yeast strains and species by immobilization in polyvinyl alcohol (PVA) hydrogel particles. RESULTS: Debaryomyces hansenii, Pichia sorbitophila, Saccharomyces cerevisiae, Yarrowia lipolytica, and Zygosaccharomyces rouxii were immobilized in PVA particles using LentiKats technology and stored in sterile water at 4 °C. The immobilization improved the survival of all species; however, the highest storage stability was achieved for S...
November 30, 2017: Biotechnology Letters
Lynn Wong, Jake Engel, Erqing Jin, Benjamin Holdridge, Peng Xu
Effective metabolic engineering of microorganisms relies on balanced expression of both heterologous and endogenous genes to channel metabolic flux towards products of interest while achieving reasonable biomass buildup. To facilitate combinatorial pathway engineering and facile genetic operation, we engineered a set of modular cloning vectors compatible with BioBrick standards, called YaliBricks, to allow for rapid assembly of multigene pathways with customized genetic control elements (promoters, intronic sequences and terminators) in the oleaginous yeast Yarrowia lipolytica...
December 2017: Metabolic Engineering Communications
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