Methanol, a rich one-carbon feedstock, can be massively produced from CO2 by the liquid sunshine route, which is helpful to realize carbon neutrality. β-Farnesene is widely used in the production of polymers, surfactants, lubricants, and also serves as a suitable substitute for jet fuel. Constructing an efficient cell factory is a feasible approach for β-farnesene production through methanol biotransformation. Here, we extensively engineered the methylotrophic yeast Ogataea polymorpha for the efficient bio-production of β-farnesene using methanol as the sole carbon source. Our study demonstrated that sufficient supply of precursor acetyl-CoA and cofactor NADPH in an excellent yeast chassis had a 1.3-fold higher β-farnesene production than that of wild-type background strain. Further optimization of the mevalonate pathway and enhancement of acetyl-CoA supply led to a 7-fold increase in β-farnesene accumulation, achieving the highest reported sesquiterpenoids production (14.7 g/L with a yield of 46 mg/g methanol) from one-carbon feedstock under fed-batch fermentation in bioreactor. This study demonstrates the great potential of engineering O. polymorpha for high-level terpenoid production from methanol.

中文翻译：

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用于从单一甲醇中高水平生产 β-法呢烯的工程酵母


甲醇是一种丰富的一碳原料，可以通过液态阳光路线从 CO2 中大规模生产，有助于实现碳中和。β-法呢烯广泛用于聚合物、表面活性剂、润滑剂的生产，也可用作喷气燃料的合适替代品。构建高效的细胞工厂是通过甲醇生物转化生产 β-法呢烯的可行方法。在这里，我们广泛设计了甲基营养酵母 Ogataea polymorpha，用于使用甲醇作为唯一的碳源高效生物生产 β-法呢烯。我们的研究表明，在优良的酵母底盘中，前体乙酰辅酶 A 和辅因子 NADPH 的充足供应比野生型背景菌株的 β-法呢烯产量高 1.3 倍。进一步优化甲羟戊酸途径和增强乙酰辅酶 A 供应导致 β-法呢烯积累增加 7 倍，在生物反应器中补料分批发酵下，单碳原料实现了最高的倍半萜类化合物产量（14.7 g/L，产率为 46 mg/g 甲醇）。本研究证明了工程化 O. polymorpha 在甲醇中高水平萜类化合物生产的巨大潜力。