Enhanced Isoprene Production by Reconstruction of Metabolic Balance between Strengthened Precursor Supply and Improved Isoprene Synthase in Saccharomyces cerevisiae,ACS Synthetic Biology

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Enhanced Isoprene Production by Reconstruction of Metabolic Balance between Strengthened Precursor Supply and Improved Isoprene Synthase in Saccharomyces cerevisiae
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-08-25 00:00:00 , DOI: 10.1021/acssynbio.8b00289
Zhen Yao ₁ , Pingping Zhou ₁ , Bingmei Su ₂ , Sisi Su ₃ , Lidan Ye _{1,

4} , Hongwei Yu _{1,

4}

Affiliation

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.

中文翻译：

通过在酿酒酵母中增强前体供应与改善的异戊二烯合酶之间的代谢平衡来增强异戊二烯的生产

异戊二烯作为一种通用的散装化学品，具有广泛的工业应用。在这里，我们试图提高异戊二烯合成酵母通过前体供应和转换的同时加强通过途径分隔和蛋白质工程的结合。首先，通过饱和诱变产生了优异的异戊二烯合酶突变体ISPSLN，导致异戊二烯产量提高了近4倍。随后将ISPSLN引入细胞质或线粒体中具有增强前体供应的菌株，这意味着异戊烯基焦磷酸酯（IPP）/二甲基烯丙基二磷酸酯（DMAPP）库的合成与转化之间存在不完全匹配。为了重建上游和下游通量之间的代谢平衡，可额外复制二磷酸戊戊二酸脱羧酶基因（MVD1）和异戊烯基-二磷酸δ-异构酶基因（IDI1））被引入细胞质和线粒体工程菌株中。最后，通过将上述单倍体菌株交配而产生的二倍体菌株产生了11.9 g / L的异戊二烯，这是真核细胞中报道的最高值。

更新日期：2018-08-25

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