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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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.