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结合蛋白质和有机体工程,用于Saccharomyces cerevisiae中的林纳过度生产

Pingping Zhou^1,2, Xiuqi Zhou², Doudou Yuan²

¹Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Yangzhou, Jiangsu 225009, PR China.

Journal of agricultural and food chemistry

|June 23, 2023

概括

通过对林纳醇合成酶进行工程并将其在酵母中的合成细分,提高了微生物的林纳醇生产. 这实现了创纪录的产量,为生产这种有价值的单烯提供了一个可持续的替代方案.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
代谢工程是代谢工程.

背景情况:

林纳是一种有价值的单烯,对香水,化品和制药行业至关重要.
微生物生产为植物提取和化学合成提供了一个可持续的替代方案.
低酶活性和前体可用性限制了当前微生物林纳醇产量.

研究的目的:

通过蛋白质工程和途径优化,增强Saccharomyces cerevisiae中的林纳醇生产.
克服微生物林纳醇合成的催化效率和前体供应的局限性.

主要方法:

通过修改基质结合口袋入口 (F447E和F447A突变) 来合理设计林纳醇合成酶 (t67OMcLIS_M).
在细胞质和过氧体中生物合成途径的分区.
微生物菌株的优化,以通过摇瓶和料批发发酵来生产高度林纳醇.

主要成果:

改进后的F447E和F447A变种显示出更高的催化效率,分别提高了2.2倍和1.9倍的林纳醇产量.
在使用工程酵母的摇瓶种植中,获得了219.1 mg/L的高标位.
在5升料批发发酵中产生了创纪录的2.6g/L的林纳醇,这是关于酵母的最高报告.

结论:

林纳醇合成酶的蛋白质工程显著提高了催化效率.
路径分隔,特别是在过氧体中,增强了微生物的林纳醇生产.
这项研究为微生物合成单烯提供了坚实的框架.

关键词:

这种植物是Saccharomyces cerevisiae.林纳醇合成酶的使用方法单烯单烯的使用方法过氧化物体是多氧化物体蛋白质工程工程蛋白质工程

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