Engineering central metabolism in Yarrowia lipolytica increases lipid accumulation

IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biochemical Engineering Journal Pub Date : 2024-11-22 DOI:10.1016/j.bej.2024.109589
Tadej Markuš , Mladen Soldat , Vasilka Magdevska , Jaka Horvat , Martin Kavšček , Gregor Kosec , Štefan Fujs , Uroš Petrovič
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Abstract

Yarrowia lipolytica, a non-conventional oleaginous yeast, holds great promise for a wide range of metabolic engineering applications. Strains of this species are renowned for their capacity to accommodate high carbon flux through acetyl-CoA and are, therefore, being extensively studied for producing chemicals derived from this precursor molecule. We investigated the effects of the pyruvate bypass, phosphoketolase bypass, and the carnitine shuttle on Y. lipolytica’s central carbon metabolism by monitoring the production of lipids and β-carotene. We constructed a set of parental strains derived from the Y. lipolytica YB-392 wild-type strain, introduced the metabolic bypasses, and subjected the constructed strains to a series of fermentation experiments. We discovered that upon introducing the modifications to central carbon metabolism, carbon is preferentially directed through the lipid biosynthesis pathway, making such engineering an effective strategy for increasing lipid production. The highest lipid content of 0.61 glipids/gDCW was recorded after introducing the carnitine shuttle-associated gene YlCAT2. Meanwhile, the highest increase in production efficiency, 78.2 %, was recorded for a combination of genes associated with the pyruvate bypass (YlPDC1, YlALD3, YlACS1).
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脂肪溶解亚罗菌的中枢代谢工程可增加脂质积累
脂溶性亚罗酵母(Yarrowia lipolytica)是一种非常规的含油酵母,在广泛的代谢工程应用中大有可为。该菌株以其通过乙酰-CoA 来适应高碳通量的能力而闻名,因此被广泛研究用于生产由这种前体分子衍生的化学物质。我们通过监测脂质和β-胡萝卜素的生产,研究了丙酮酸旁路、磷酸酮酶旁路和肉碱穿梭对溶脂酵母的中心碳代谢的影响。我们构建了一组源自脂溶性酵母 YB-392 野生型菌株的亲本菌株,引入了代谢旁路,并对所构建的菌株进行了一系列发酵实验。我们发现,在引入对中心碳代谢的改造后,碳会优先通过脂质生物合成途径,从而使这种工程改造成为提高脂质产量的有效策略。引入肉碱穿梭相关基因 YlCAT2 后,脂质含量最高,达到 0.61g/gDCW。同时,与丙酮酸旁路相关的基因组合(YlPDC1、YlALD3、YlACS1)的生产效率增幅最高,达到 78.2%。
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来源期刊
Biochemical Engineering Journal
Biochemical Engineering Journal 工程技术-工程:化工
CiteScore
7.10
自引率
5.10%
发文量
380
审稿时长
34 days
期刊介绍: The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology. The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields: Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics Biosensors and Biodevices including biofabrication and novel fuel cell development Bioseparations including scale-up and protein refolding/renaturation Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells Bioreactor Systems including characterization, optimization and scale-up Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis Protein Engineering including enzyme engineering and directed evolution.
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