Metabolic engineering of Escherichia coli for efficient production of (3R)-acetoin
Metabolic engineering of Escherichia coli for efficient production of (3R)-acetoin
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DOI：
10.1002/jctb.4293
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发表时间：
2015-01-01
影响因子：
3.4
通讯作者：
Zhang, Liaoyuan
中科院分区：
文献类型：
--
作者：
Xu, Quanming;Xie, Linxiang;Zhang, Liaoyuan
光学纯乙偶姻是一种重要的潜在的医药中间体。它也被广泛用于合成新型光学活性的β-羟基酮衍生物和液晶复合物。重组大肠杆菌被开发用于高效生产（3R）-乙偶姻。通过对工程菌的培养基优化和工艺控制，提高了（3R）-乙偶姻的产量。大肠杆菌中高效生产（3R）-乙偶姻。分批培养结果表明，该工程菌从60 g L-1葡萄糖中可获得23.4 g L-1的（3R）-乙偶姻。手性柱GC分析表明，所产生的（3R）-乙偶姻的立体异构体纯度为97.3%。采用正交设计法对摇瓶培养基组成进行了优化。在最佳条件下摇瓶发酵，（3R）-乙偶姻的产量可达38.3 g L-1。在5L的生物反应器中，在合适的搅拌速度下进行流加发酵，最高（3R）-乙偶姻浓度为60.3g L-1，生产能力为1.55g L-1h（-1），产率为86.3%。构建了高效生产（3R）-乙偶姻的大肠杆菌。发酵变量和补料分批培养的优化导致最大（3R）-乙偶姻浓度为60.3 g L-1。(c)2013年化学工业协会
BACKGROUNDOptically pure acetoin is an important potential pharmaceutical intermediate. It has also been widely used to synthesize novel optically active -hydroxyketone derivatives and liquid crystal composites. Recombinant Escherichia coli was developed for efficient (3R)-acetoin production. Culture medium optimization and process control were carried out to improve (3R)-acetoin yield by the engineered strain.RESULTSA synthetic pathway involved the budRAB genes from Serratia marcescens and NADH oxidase gene from Lactobacillus brevis in E. coli was developed for efficient (3R)-acetoin production. Batch culture showed that 23.4 g L-1 of (3R)-acetoin could be obtained from 60 g L-1 glucose by the engineered strain. Chiral-column GC analysis indicated that the stereoisomeric purity of (3R)-acetoin produced was 97.3%. Further, the medium composition was optimized in shake flasks by an orthogonal design method. Under optimal conditions, (3R)-acetoin concentration reached 38.3 g L-1 in flask fermentation. Fed-batch fermentation based on a suitable agitation speed was carried out in a 5 L bioreactor, and maximum (3R)-acetoin concentration of 60.3 g L-1 was achieved with a productivity of 1.55 g L-1 h(-1) and yield 86.3%.CONCLUSIONAn engineering E. coli for efficient (3R)-acetoin production was constructed. The optimization of fermentation variables and fed-batch culture resulted in a maximum (3R)-acetoin concentration of 60.3 g L-1. (c) 2013 Society of Chemical Industry