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代谢工程改造酿酒酵母生产次丹参酮二烯
首发时间：2025-03-13
摘要：次丹参酮二烯是合成有药用价值的二萜类化合物的关键中间体。在本研改造了酿酒酵母以实现次丹参酮二烯的高产，通过下调erg9基因，过表达thmg1和bts1-erg20基因来增强mva途径，并引入saggps基因，增加前体物质ggpp供应。随后，通过增加saggps、thmg1基因拷贝数策略及多拷贝delta位点整合次丹参酮二烯合酶基因tskml1-cftps1策略，研究了关键基因的拷贝数对次丹参酮二烯产量的影响。结果表明，thmg1基因三个拷贝数次丹参酮二烯的产量最佳，saggps基因两个拷贝数和三个拷贝数次丹参酮二烯的产量下降，最后通过多拷贝位点优化次丹参酮二烯合酶，成功筛选到了一株次丹参酮二烯产的高产菌株，次丹参酮二烯产量达到了349.76 mg/l。这一成果为其他二萜类化合物的微生物合成提供了理论基础。
关键词： 酿酒酵母
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metabolic engineering saccharomyces cerevisiae to produce miltiradiene
abstract：miltiradiene is a key intermediate for the synthesis of medicinal diterpenoids. in this study, saccharomyces cerevisiae was transformed to achieve high yield of miltiradiene. the mva pathway was enhanced by down-regulating erg9 gene, overexpressing thmg1 and bts1-erg20 genes, and saggps gene was introduced to increase the supply of precursor ggpp. subsequently, by increasing the copy number of saggps and thmg1 genes and integrating miltiradiene synthase gene tskml1-cftps1 with multiple copies of delta site, the influence of the copy number of key genes on the yield of miltiradiene was studied. the results showed that the yield of miltiradiene with three copies of thmg1 gene was the bemetabolic engineering saccharomyces cerevisiae to produce miltiradienest, while the metabolic engineering transformation of smetabolic engineering saccharomyces cerevisiae to produce miltiradieneaccharomyces cerevisiae to produce miltiradieneyield of miltiradiene with two copies and three copies of saggps gene decreased. finally, a strain with high yield of miltiradiene was successfully screened by optimizing the miltiradiene synthase with multiple copies, and the yield of miltiradiene reached 349.76 mg/l. this achievement provides a theoretical basis for the microbial synthesis of other diterpenoids.
keywords： saccharomyces cerevisiae
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代谢工程改造酿酒酵母生产次丹参酮二烯
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