谷氨酸棒杆菌
代谢工程
羟基烷酸
合成生物学
生物化学
化学
代谢途径
工业微生物学
新陈代谢
计算生物学
细菌
生物
基因
发酵
遗传学
作者
Sung Sun Yim,Jae Woong Choi,Yong Jae Lee,Ki Jun Jeong
出处
期刊:Research Square - Research Square
日期:2022-10-12
标识
DOI:10.21203/rs.3.rs-2140595/v1
摘要
Abstract Background: The disposal of plastic waste is a major environmental challenge. With recent advances in microbial genetic and metabolic engineering technologies, microbial polyhydroxyalkanoates (PHAs) are being used as next-generation biomaterials to replace petroleum-based synthetic plastics in a sustainable future. However, the relatively high production cost of bioprocesses hinders the production and application of microbial PHAs on an industrial scale. Results: Here, we describe a rapid strategy to rewire metabolic networks in an industrial microorganism, Corynebacterium glutamicum , for the enhanced production of poly(3-hydroxybutyrate) (PHB). A three-gene PHB biosynthetic pathway in Rasltonia eutropha was refactored for high-level gene expression. A fluorescence-based quantification assay for cellular PHB content using BODIPY was devised for the rapid fluorescence-activated cell sorting (FACS)-based screening of a large combinatorial metabolic network library constructed in C. glutamicum . Rewiring metabolic networks across the central carbon metabolism enabled highly efficient production of PHB up to 29% of dry cell weight, which is the highest PHB content ever reported in C. glutamicum using a sole carbon source. Conclusions: We successfully constructed a heterologous PHB biosynthetic pathway and rapidly optimized metabolic networks across central metabolism in C. glutamicum for enhanced production of PHB using glucose or fructose as the sole carbon source in minimal medium. We expect that this FACS-based metabolic rewiring framework will accelerate strain engineering processes for the production of diverse biochemicals and biopolymers.
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