提高酵母中 8-羟基香叶醇产量的系统工程,Journal of Agricultural and Food Chemistry

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提高酵母中 8-羟基香叶醇产量的系统工程
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2023-03-01 , DOI: 10.1021/acs.jafc.2c09028
Herong Wang ₁ , Guozhen Jiang ₁ , Nan Liang ₁ , Tianyu Dong ₁ , Mengying Shan ₁ , Mingdong Yao ₁ , Ying Wang ₁ , Wenhai Xiao _{1,

2} , Yingjin Yuan ₁

Affiliation

8-羟基香叶醇是昆虫性信息素和防御性分泌物的重要组成部分，可作为潜在的农业生物驱虫剂。微生物生产为高效获得8-羟基香叶醇提供了可持续和绿色的手段。P450 geraniol-8-hydroxylase (G8H) 将 geraniol 转化为 8-hydroxygeraniol 被认为是 8-hydroxygeraniol 生产的瓶颈。在此，实施了由 G8H 和细胞色素 P450 还原酶 (CPR)、内质网 (ER) 工程和减少的烟酰胺腺嘌呤二核苷酸磷酸 (NADPH) 供应之间的适应性组成的综合策略，以提高酿酒酵母中 8-羟基香叶醇的产量. 8-羟基香叶醇的滴度逐渐增加 2.1 倍（高达 158.1 mg/L）。此外，负责将 8-羟基香叶醇还原为分流产物的脱氢酶ADH6和还原酶ARI1也被删除，将摇瓶水平的 8-羟基香叶醇产量提高至 238.9 mg/L。因此，通过碳限制策略，在 5.0 L 补料分批发酵中，酿酒酵母中的 8-羟基香叶醇含量超过 1.0 g/L ，这是迄今为止微生物中报道的最高滴度。我们的工作不仅为8-羟基香叶醇的从头生物合成提供了可持续的途径，而且为微生物 P450 工程提供了很好的参考。

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Systematic Engineering to Enhance 8-Hydroxygeraniol Production in Yeast

8-Hydroxygeraniol, an important component of insect sex pheromones and defensive secretions, can be used as a potential biological insect repellent in agriculture. Microbial production provides sustainable and green means to efficiently gain 8-hydroxygeraniol. The conversion of geraniol to 8-hydroxygeraniol by P450 geraniol-8-hydroxylase (G8H) was regarded as the bottleneck for 8-hydroxygeraniol production. Herein, an integrated strategy consisting of the fitness between G8H and cytochrome P450 reductase (CPR), endoplasmic reticulum (ER) engineering, and reduced nicotinamide adenine dinucleotide phosphate (NADPH) supply is implemented to enhance the production of 8-hydroxygeraniol in Saccharomyces cerevisiae. The titer of 8-hydroxygeraniol was gradually increased by 2.1-fold (up to 158.1 mg/L). Moreover, dehydrogenase ADH6 and reductase ARI1 responsible for the reduction of 8-hydroxygeraniol toward shunt products were also deleted, elevating 8-hydroxygeraniol production to 238.9 mg/L at the shake flask level. Consequently, more than 1.0 g/L 8-hydroxygeraniol in S. cerevisiae was achieved in 5.0 L fed-batch fermentation by a carbon restriction strategy, which was the highest-reported titer in microbes so far. Our work not only provides a sustainable way for de novo biosynthesis of 8-hydroxygeraniol but also sets a good reference in P450 engineering in microbes.

更新日期：2023-03-01

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