Systematic Engineering of Escherichia coli for D-Lactate Production from Crude Glycerol

被引:19
作者
Wang, Zei Wen [1 ]
Saini, Mukesh [1 ]
Lin, Li-Jen [2 ]
Chiang, Chung-Jen [3 ]
Chao, Yun-Peng [1 ,4 ,5 ]
机构
[1] Feng Chia Univ, Dept Chem Engn, Taichung 40724, Taiwan
[2] China Med Univ, Coll Chinese Med, Sch Chinese Med, Taichung 40402, Taiwan
[3] China Med Univ, Dept Med Lab Sci & Biotechnol, Taichung 40402, Taiwan
[4] Asia Univ, Dept Hlth & Nutr Biotechnol, Taichung 41354, Taiwan
[5] China Med Univ Hosp, Dept Med Res, Taichung 40447, Taiwan
关键词
crude glycerol; D-lactate; metabolic engineering; fermentation technology; LACTIC-ACID PRODUCTION; PURE D-LACTATE; ANAEROBIC FERMENTATION; METABOLISM; BIODIESEL; PATHWAYS; STRAINS;
D O I
10.1021/acs.jafc.5b04162
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
Crude glycerol resulting from biodiesel production is an abundant and renewable resource. However, the impurities in crude glycerol usually make microbial fermentation problematic. This issue was addressed by systematic engineering of Escherichia coli for the production of D-lactate from crude glycerol. First, mgsA and the synthetic pathways of undesired products were eliminated in E. coli, rendering the strain capable of homofermentative production of optically pure D-lactate. To direct carbon flux toward D-lactate, the resulting strain was endowed with an enhanced expression of glpD-glpK in the glycerol catabolism and of a heterologous gene encoding D-lactate dehydrogenase. Moreover, the strain was evolved to improve its utilization of cruder glycerol and subsequently equipped with the FocA channel to export intracellular D-lactate. Finally, the fed-batch fermentation with two-phase culturing was carried out with a bioreactor. As a result, the engineered strain enabled production of 105 g/L D-lactate (99.9% optical purity) from 121 g/L crude glycerol at 40 h. The result indicates the feasibility of our approach to engineering E. coli for the crude glycerol-based fermentation.
引用
收藏
页码:9583 / 9589
页数:7
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