Genetic engineering and production of modified fatty acids by the non-conventional oleaginous yeast Trichosporon oleaginosus ATCC 20509

Görner, Christian; Redai, Veronika; Bracharz, Felix; Schrepfer, Patrick; Garbe, Daniel; Brück, Thomas

doi:10.1039/c5gc01767j

Cited by 63 publications

(67 citation statements)

References 46 publications

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“…Recently, the review of Yaguchi et al [ 6 ], made an excellent effort to summarize and contrast the data for the prominent oleaginous yeasts C. oleaginosus and Debaryomyces hansenii . This review aims to further extend on available data of C. oleaginosus to provide the reader a comprehensive but focused overview of the metabolic capacity of this intriguing organism, which most recently has been rendered genetically accessible [ 7 ]. More generally, C. oleaginosus grows on a variety of complex biomass hydrolysates and even in the presence on fermentation inhibitors.…”

Section: Introductionmentioning

confidence: 99%

Opportunities and challenges in the development of Cutaneotrichosporon oleaginosus ATCC 20509 as a new cell factory for custom tailored microbial oils

et al. 2017

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Cutaneotrichosporon oleaginosus ATCC 20509, previously known as Trichosporon oleaginosus, Cryptococcus curvatus, Apiotrichum curvatum or Candida curvata D is an oleaginous yeast with several favorable qualities: it is fast growing, accumulates high amounts of lipid and has a very broad substrate spectrum. Its resistance to hydrolysis byproducts and genetic accessibility make it a promising cell factory for custom tailored microbial oils. However, literature about this organism is of varying degree of quality. Moreover, due to numerous changes of the species name, reports are highly scattered and poorly cited. This led to a poor integration of the findings into a unified body of knowledge. Particularly, errors in strain name usage and consequently citation are found even in most recent literature. To simplify future work, this review provides an overview of published studies and main findings regarding the metabolic capacities of C. oleaginosus.

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Section: Introductionmentioning

confidence: 99%

Opportunities and challenges in the development of Cutaneotrichosporon oleaginosus ATCC 20509 as a new cell factory for custom tailored microbial oils

et al. 2017

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“…According to [25], comparing fructose, glucose, xylose and sucrose, C. oleaginosus ATCC 20509 grows the fastest in fructose, the slowest in sucrose while there is no significant difference between glucose or xylose. According to [26] xylose is favored over glucose for biomass generation. These differences can be due to various factors such as pH, temperature, oxygen, dilution rate, and fermentation modes across experiments.…”

Section: Effect Of the C/n Ratio On Lipid Productionmentioning

confidence: 99%

“…lipid production], is typically a multi-factorial process depending on the growth medium, culture conditions, strain specificity and the interplay among these factors. Hence, a predictive constraintbased, genome-scale model of metabolism (GEM), along with genetic accessibility tools [26] will provide new avenues towards reaching the full potential of C. oleaginosus ATCC 20509 as a lipid producer [11].…”

Section: Introductionmentioning

confidence: 99%

Genome-scale metabolic modelling underscores the potential of Cutaneotrichosporon oleaginosus ATCC 20509 as a cell factory for biofuel production

Pham

Reijnders

Suárez-Diez

et al. 2020

Preprint

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Abstract Background: Cutaneotrichosporon oleaginosus ATCC 20509 is a fast growing oleaginous basidiomycete yeast that is able to grow in a wide range of low-cost carbon sources including crude glycerol, a byproduct of biodiesel production. When glycerol is used as a carbon source, this yeast can accumulate more than 50% lipids (w/w) with high concentrations of mono-unsaturated fatty acids. Results: To increase our understanding of this yeast and to provide a knowledge base for further industrial use, a FAIR re-annotated genome was used to build a genome-scale, constraint-based metabolic model containing 1553 reactions involving 1373 metabolites in 11 compartments. A new description of the biomass synthesis reaction was introduced to account for massive lipid accumulation in conditions with high carbon to nitrogen (C/N) ratio in the media. This condition-speciﬁc biomass objective function is shown to better predict conditions with high lipid accumulation using glucose, fructose, sucrose, xylose and glycerol as sole carbon source. Conclusion: Contributing to the economic viability of biodiesel as renewable fuel, C. oleaginosus ATCC 20509 can eﬀectively convert crude glycerol waste streams in lipids as a potential bioenergy source. Performance simulations are essential to identify optimal production conditions and to develop and ﬁne tune a cost-eﬀective production process. Our model suggests ATP-citrate lyase as a target for further improve lipid production. Keywords: Genome-scale metabolic model; Cutaneotrichosporon oleaginosus ATCC 20509; lipid accumulation; Crude glycerol; biodiesel production; ﬂux balance analysis; oleaginous yeast

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“…While lytic enzymes infer a more gentle and effective disintegration alternative, they were eliminated from this study as they impose additional downstream processing in proteomic exploration (removal of lytic enzymes and/or preparation of laborious MS exclusion lists) [53,54]. Notably, pelleting of C. oleaginosus, for media elimination and washing, necessitated the use of 40% v/v ethanol to avoid loss of "cellular floaters" that result from high lipid content.…”

Section: Lysis Efficiencymentioning

confidence: 99%

Optimization of protein isolation by proteomic qualification from Cutaneotrichosporon oleaginosus

Awad

Brück

2019

Anal Bioanal Chem

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In the last decades, microbial oils have been extensively investigated as a renewable platform for biofuel and oleochemical production. Offering a potent alternative to plant-based oils, oleaginous microorganisms have been the target of ongoing metabolic engineering aimed at increasing growth and lipid yields, in addition to specialty fatty acids. Discovery proteomics is an attractive tool for elucidating lipogenesis and identifying metabolic bottlenecks, feedback regulation, and competing biosynthetic pathways. One prominent microbial oil producer is Cutaneotrichosporon oleaginosus, due to its broad feedstock catabolism and high lipid yield. However, this yeast has a recalcitrant cell wall and high cell lipid content, which complicates efficient and unbiased protein extraction for downstream proteomic analysis. Optimization efforts of protein sample preparation from C. oleaginosus in the present study encompasses the comparison of 8 lysis methods, 13 extraction buffers, and 17 purification methods with respect to protein abundance, proteome coverage, applicability, and physiochemical properties (pI, MW, hydrophobicity in addition to COG, and GO analysis). The optimized protocol presented in this work entails a one-step extraction method utilizing an optimal lysis method (liquid homogenization), which is augmented with a superior extraction buffer (50 mM Tris, 8/2 M Urea/Thiourea, and 1% C7BzO), followed by either of 2 advantageous purification methods (hexane/ ethanol or TCA/acetone), depending on subsequent applications and target studies. This work presents a significant step forward towards implementation of efficient C. oleaginosus proteome mining for the identification of potential targets for genetic optimization of this yeast to improve lipogenesis and production of specialty lipids.

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Genetic engineering and production of modified fatty acids by the non-conventional oleaginous yeast Trichosporon oleaginosus ATCC 20509

Abstract: Establishing genetic engineering tools for sustainable production of tailor made fatty acids in the non-conventional, oleaginous yeast Trichosporon oleaginosus.

Cited by 63 publications

References 46 publications

Opportunities and challenges in the development of Cutaneotrichosporon oleaginosus ATCC 20509 as a new cell factory for custom tailored microbial oils

Opportunities and challenges in the development of Cutaneotrichosporon oleaginosus ATCC 20509 as a new cell factory for custom tailored microbial oils

Genome-scale metabolic modelling underscores the potential of Cutaneotrichosporon oleaginosus ATCC 20509 as a cell factory for biofuel production

Optimization of protein isolation by proteomic qualification from Cutaneotrichosporon oleaginosus

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