Engineering Pichia pastoris for high-level biosynthesis of squalene

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2025-02-19 DOI:10.1016/j.bej.2025.109677

Xinying Zhang , Yuxin Yang , Shuting Chen , Ying Lin , Yanxuan Wu , Shuli Liang

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引用次数: 0

Abstract

High-value-added chemicals and advanced biofuels can be feasibly produced from renewable feedstocks through microbial metabolic engineering. Methanol is produced with high quantities from CO₂ using green energy sources such as solar energy, thereby making it an ideal feedstock. However, little has been documented on the microbial production of terpenoids utilising methanol. This study aimed to engineer Pichia pastoris to over-produce triterpene squalene from methanol by optimising and reconstructing the utilisation pathways of mevalonate and methanol. In addition, we explored the exocytosis of squalene and promoted squalene secretion successfully. Furthermore, we identified a key bottleneck of squalene synthesis in P. pastoris through transcriptomic analysis. The final engineered strain produced 20.80 ± 0.02 g/L squalene during fed-batch fermentation, which is the highest squalene production using organic one-carbon as carbon source reported so far. Our findings set the stage for applying P. pastoris as an organic one-carbon platform for producing biochemicals and biofuels.

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工程毕赤酵母高水平生物合成角鲨烯

通过微生物代谢工程，利用可再生原料生产高附加值化学品和先进生物燃料是可行的。甲醇是利用太阳能等绿色能源从二氧化碳中大量生产出来的，因此它是一种理想的原料。然而，很少有文献记载的微生物生产萜类化合物利用甲醇。本研究旨在通过优化和重构甲戊酸盐和甲醇的利用途径，设计毕赤酵母从甲醇中过量生产三萜角鲨烯。此外，我们还探索了角鲨烯的胞吐作用，成功地促进了角鲨烯的分泌。此外，我们通过转录组学分析确定了巴斯德酵母中角鲨烯合成的关键瓶颈。最终工程菌株补料间歇发酵产鲨烯20.80 ± 0.02 g/L，是目前报道的以有机一碳为碳源产鲨烯量最高的菌株。我们的发现为将巴氏酵母应用于生产生物化学品和生物燃料的有机单碳平台奠定了基础。

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来源期刊

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.