A systematic optimization of medium chain fatty acid biosynthesis via the reverse beta-oxidation cycle in Escherichia coli

文献类型： 外文期刊

第一作者： Wu, Junjun

作者： Wu, Junjun;Zhang, Xia;Dong, Mingsheng;Xia, Xiudong

作者机构：

关键词： beta-oxidation reversal;Biofuels;Biochemical;Synthetic biology;Metabolic engineering

期刊名称：METABOLIC ENGINEERING （ 影响因子：9.783； 五年影响因子：9.566 ）

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收录情况： SCI

摘要： Medium-chain fatty acids (MCFAs, 6-10 carbons) are valuable precursors to many industrial biofuels and chemicals, recently engineered reversal of the beta-oxidation (r-BOX) cycle has been proposed as a potential platform for efficient synthesis of MCFAs. Previous studies have made many exciting achievements on functionally characterizing four core enzymes of this r-BOX cycle. However, the information about bottleneck nodes in this cycle is elusive. Here, a quantitative assessment of the inherent limitations of this cycle was conducted to capitalize on its potential. The selection of the core beta-oxidation reversal enzymes in conjunction with acetyl-CoA synthetase endowed the ability to synthesize about 1 g/L MCFAs. Furthermore, a gene dosage experiment was developed to identify two rate-limiting enzymes (acetyl-CoA synthetase and thiolase). The de novo pathway was then separated into two modules at thiolase and MCFA production titer increased to 2.8 g/L after evaluating different construct environments. Additionally, the metabolism of host organism was reprogrammed to the desired biochemical product by the clustered regularly interspaced short palindromic repeats interference system, resulted in a final MCFA production of 3.8 g/L. These findings described here identified the inherent limitations of r-BOX cycle and further unleashed the lipogenic potential of this cycle, thus paving the way for the development of a bacterial platform for microbial production of high-value oleochemicals from low-value carbons in a sustainable and environmentally friendly manner.

分类号： Q51

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