酿酒酵母异丁醇合成途径调控的研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2022-1350

摘要/Abstract

摘要：

酿酒酵母是工业化生产中常用的微生物，具有耐低pH值、抗逆性强等优点。利用微生物进行生物基产品的生产已成为绿色生物制造的重大发展方向。异丁醇是一种支链高级醇，在化工、能源等领域有较好的应用前景。利用工程酿酒酵母合成的异丁醇是一种可持续、可再生、经济环保的生物能源，有望成为下一代生物燃料。构建高效合成异丁醇的酿酒酵母细胞工厂可以实现生物燃料异丁醇的绿色制造，但目前酿酒酵母合成异丁醇的能力不足，限制了异丁醇的工业化发展，而与合成生物学相结合的理性代谢调控策略是提高微生物异丁醇合成能力的有效方法之一。本文围绕异丁醇合成途径构建、竞争途径阻断、辅因子平衡、合成途径重定位、转录调控因子调节以及耐受性强化几方面，综述了酿酒酵母中异丁醇合成调控策略的研究进展。这些策略显著提高了工程酵母异丁醇的转化率和产率，但在异丁醇工业化生产上仍存在许多不足。为了推动异丁醇的工业化生产，本文讨论了目前酿酒酵母生产异丁醇的瓶颈及解决方法，以期对异丁醇的绿色制造提供有价值的参考。

关键词: 酿酒酵母, 异丁醇, 生物能源, 代谢调控, 合成生物学

Abstract:

As one of the microorganism commonly used in industrial production, Saccharomyces cerevisiae is of low pH resistance and strong stress resistance. The use of microorganisms to produce bio-based products has become an important development direction of green biomanufacturing. Isobutanol is a kind of branched-chain alcohol, which has a good application prospect in the chemical industry, energy, and other fields. As a biofuel, the synthesized isobutanol from S. cerevisiae is a sustainable, renewable, economical and environment-friendly approach to meet the needs of human development. The construction of an efficient isobutanol synthesizing S. cerevisiae cell factory may enable the green manufacturing of isobutanol as a biofuel. However, the ability of S. cerevisiae to synthesize isobutanol has limited the industrial development of isobutanol, the rational metabolic regulation strategy combined with synthetic biology is one of the effective methods to improve the ability of microbial isobutanol synthesis. This review summarized the latest developments in the regulating strategies of isobutanol biosynthesis in S. cerevisiae, including promoting isobutanol synthesis pathways, blocking competitive pathways, balancing cofactors, relocating synthetic pathways, regulating transcription factors, and improving microbial isobutanol tolerance. These strategies have significantly improved the conversion and yield of isobutanol in engineering yeast, but there are still many deficiencies in the industrial production of isobutanol. In order to promote the industrial production of isobutanol, the current bottlenecks and their solutions for isobutanol production in S. cerevisiae were discussed, aiming to provide valuable information for the green manufacturing of isobutanol.

Key words: Saccharomyces cerevisiae, isobutanol, biofuel, metabolic regulation, synthetic biology

成婷, 苑帅, 张晓元, 林良才, 李欣, 张翠英. 酿酒酵母异丁醇合成途径调控的研究进展[J]. 生物技术通报, 2023, 39(7): 80-90.

CHENG Ting, YUAN Shuai, ZHANG Xiao-yuan, LIN Liang-cai, LI Xin, ZHANG Cui-ying. Research Progress in the Regulation of Isobutanol Synthesis Pathway in Saccharomyces cerevisiae[J]. Biotechnology Bulletin, 2023, 39(7): 80-90.

图/表 3

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Strain background	Pathway localization	Cofactor-balance	Overexpressing isobutanol synthesis pathway	Block isobutanol competing pathway	Transcription factor regulation	Improved isobutanol tolerance	Isobutanol titer /（mg·L^-1）	Isobutanol yield/ （mg·g^-1 carbon source）	Carbon source/Medium properties/Fermentation conditions	Reference
YPH499			√	√			143.0	6.6	Glucose/defined/microaerbic	[15]
CEN.PK2-1C			√	√	√		376.9	*	Glucose/defined/aerobic	[29]
YPH499		√	√	√			224.0	12.0	Glucose/defined/aerobic	[20]
CEN.PK2-1C	√			√			330.9	*	Glucose/defined/aerobic	[25]
CEN.PK2-1C		√	√	√	√		1 245.0	12.5	Glucose/defined/aerobic	[37]
D452-2	√	√		√			662.0	6.7	Glucose/defined/microaerbic	[22]
CEN.PK113-7D	√			√			2 090.0	59.6	Glucose/defined/aerobic	[1]
CEN.PK2-1C	√			√	√		263.2	*	Glucose/defined/aerobic	[33]
CEN.PK113-5D	√	√					190.0	*	Glucose/defined/anaerobic	[31]
D452-2	√		√				151.0	*	Glucose/complex/microaerbic	[35]
W303-1A			√			√	404.0	10.1	Glucose/complex/microaerbic	[48]
W303-1A			√		√	√	556.0	*	Glucose/complex/microaerbic	[47]
SR8	√		√				2 600.0	*	xylose/defined/aerobic	[11]
BY4742	√		√			√	92.9	*	xylose/defined/aerobic	[4]

Strain background	Pathway localization	Cofactor-balance	Overexpressing isobutanol synthesis pathway	Block isobutanol competing pathway	Transcription factor regulation	Improved isobutanol tolerance	Isobutanol titer /（mg·L^-1）	Isobutanol yield/ （mg·g^-1 carbon source）	Carbon source/Medium properties/Fermentation conditions	Reference
YPH499			√	√			143.0	6.6	Glucose/defined/microaerbic	[15]
CEN.PK2-1C			√	√	√		376.9	*	Glucose/defined/aerobic	[29]
YPH499		√	√	√			224.0	12.0	Glucose/defined/aerobic	[20]
CEN.PK2-1C	√			√			330.9	*	Glucose/defined/aerobic	[25]
CEN.PK2-1C		√	√	√	√		1 245.0	12.5	Glucose/defined/aerobic	[37]
D452-2	√	√		√			662.0	6.7	Glucose/defined/microaerbic	[22]
CEN.PK113-7D	√			√			2 090.0	59.6	Glucose/defined/aerobic	[1]
CEN.PK2-1C	√			√	√		263.2	*	Glucose/defined/aerobic	[33]
CEN.PK113-5D	√	√					190.0	*	Glucose/defined/anaerobic	[31]
D452-2	√		√				151.0	*	Glucose/complex/microaerbic	[35]
W303-1A			√			√	404.0	10.1	Glucose/complex/microaerbic	[48]
W303-1A			√		√	√	556.0	*	Glucose/complex/microaerbic	[47]
SR8	√		√				2 600.0	*	xylose/defined/aerobic	[11]
BY4742	√		√			√	92.9	*	xylose/defined/aerobic	[4]