代谢工程改造酵母生产香料的研究进展

doi:10.3969/j.issn.1001-1803.2022.06.011

摘要/Abstract

摘要：

天然产物香料作为食品、药品和日用化学品的添加成分,市场需求高,有着较高的开发利用价值。从动植物中提取香料产物存在来源少、含量低和分离困难等缺点,化学合成香料又存在着安全隐患和污染问题,很难满足市场需求。因此,研究与开发生产天然产物香料的微生物来补充或代替原有的传统提取和化学合成方法具有重要的理论意义和潜在的应用价值。酵母菌作为代谢工程菌株的一类,细胞内具有更好表达异源蛋白的细胞结构和翻译后修饰机制。文章总结了有关对酵母菌进行代谢改造生产萜类、芳香族类及脂肪族类香料化合物的具体实例和研究进展,包括所涉及的宿主菌株、关键酶、代谢途径、改造策略以及发酵条件优化等,并在最后讨论了酵母代谢工程改造生产天然产物香料目前所面临的问题和未来的发展方向。

关键词: 酵母, 代谢工程改造, 香料, 萜类, 芳香族类, 脂肪族类

Abstract:

Natural product flavors, as additives for food, medicine and daily chemicals, have high market demand and have high development and utilization value. The extraction of fragrance products from animals and plants has the disadvantages of few sources, low content and difficulty in separation. Chemically synthesized fragrances also have potential safety hazards and pollution problems, and it is difficult to meet market demand. Therefore, the research and development of microorganisms that produce natural product fragrances to supplement or replace the original traditional extraction and chemical synthesis methods has important theoretical significance and potential application value. This article mainly summarizes the specific examples and research progress related to the metabolic modification of yeasts to produce terpenoids, aromatics and aliphatic fragrance compounds, including the host strains involved, key enzymes, metabolic pathways, modification strategies, and optimization of fermentation conditions. Yeasts, as a class of metabolically engineered strains, have a cell structure and a post-translational modification mechanism for expressing heterologous proteins that other microorganisms do not possess. Therefore, compared with other microorganisms, yeast can better express heterologous proteins and has greater potential as a chassis cell for the production of natural products. This article systematically summarizes the specific examples and research progress of yeast metabolic transformation to produce terpenoids, aromatics and aliphatic spices. It is found that the competition pathway and negative feedback regulation of the target product metabolism pathway, the tolerance of the chassis cells to the target product, the supply of the target product precursor compound and the difference in the selection of the substrate by the enzyme in the synthesis pathway are the biggest problems that limit the output of the product. Furthermore, relevant suggestions and prospects are put forward for the bottleneck problems faced by the use of metabolic engineering to transform microorganisms to synthesize natural product fragrance products, aiming to provide a reference for constructing chassis yeast cells for efficient production of fragrance products.

Key words: yeast, metabolic engineering, flavors, terpenoids, aromatics, aliphatics

中图分类号:

TQ651

张永涛,赵净沙,蒋雁冰,赵丹,王昌涛,李萌. 代谢工程改造酵母生产香料的研究进展[J]. 日用化学工业, 2022, 52(6): 645-655.

Zhang Yongtao,Zhao Jingsha,Jiang Yanbing,Zhao Dan,Wang Changtao,Li Meng. Research progress in yeast metabolic engineering to produce natural product flavors[J]. China Surfactant Detergent & Cosmetics, 2022, 52(6): 645-655.

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特点	全球年均需求量	数量	香料类别
大宗常用	>5 000 t	>100	香叶醇、芳樟醇、薄荷醇等
一般常用	500~5 000 t	>300	麝香、乙偶姻等
次常用	<500 t	>6 000	硫噻唑、草莓酸、茶香酮等

菌株	萜类	工程策略	发酵条件	滴度/产率	参考
S. cerevisiae YLin-12 strain	芳樟醇（无环单萜）	SKIK tag与芳樟醇合成酶t67OMcLISE343D/E352HN端融合 SKIK-t67OMcLISE343D/E352H和ERG20F96W/N127W的组装 PERG1介导ERG20下调	补料分批发酵	80.9 mg/L	[20]
S. cerevisiae SCICK17 strain	α-檀香烯（倍半萜）	引入异源檀香烯合酶（SanSyn）过表达HMGR、下调ERG9、敲除DPP1和LPP1	补料分批发酵	92 mg/L	[21]
S. cerevisiae BY4741 strain	广藿香醇（倍半萜）	ERG20-PTS融合 HMGR,IDI1,UPC2-1过表达 EGR9启动子由HXT1替代除DPP1和LPP1	5 L补料分批生物反应器发酵	466.8 mg/L	[22]
P. pastoris	龙涎香醇（三萜）	引入AaSHC D377C和BmeTC D373C环化酶基因 HMGR和ERG9过表达加入特比萘芬抑制ERG1	补料分批发酵	14.9 mg/L	[23]