异戊二烯
酿酒酵母
生物化学
代谢工程
异构酶
饱和突变
突变体
化学
酵母
ATP合酶
淀粉体
生物
基因
叶绿体
质体
有机化学
聚合物
共聚物
作者
Zhen Yao,Pingping Zhou,Bingmei Su,Sisi Su,Lidan Ye,Hongwei Yu
标识
DOI:10.1021/acssynbio.8b00289
摘要
Isoprene, as a versatile bulk chemical, has wide industrial applications. Here, we attempted to improve isoprene biosynthesis in Saccharomyces cerevisiae by simultaneous strengthening of precursor supply and conversion via a combination of pathway compartmentation and protein engineering. At first, a superior isoprene synthase mutant ISPSLN was created by saturation mutagenesis, leading to almost 4-fold improvement in isoprene production. Subsequent introduction of ISPSLN to strains with strengthened precursor supply in either cytoplasm or mitochondria implied an imperfect match between the synthesis and conversion of the isopentenyl pyrophosphate (IPP)/dimethylallyl diphosphate (DMAPP) pool. To reconstruct metabolic balance between the upstream and downstream flux, additional copies of diphosphomevalonate decarboxylase gene (MVD1) and isopentenyl-diphosphate delta-isomerase gene (IDI1) were introduced into the cytoplasmic and mitochondrial engineered strains. Finally, the diploid strain created by mating the above haploid strains produced 11.9 g/L of isoprene, the highest ever reported in eukaryotic cells.
科研通智能强力驱动
Strongly Powered by AbleSci AI