Metabolic engineering of a Saccharomyces cerevisiae strain capable of simultaneously utilizing glucose and galactose to produce enantiopure (2R,3R)-butanediol
Metabolic engineering of a Saccharomyces cerevisiae strain capable of simultaneously utilizing glucose and galactose to produce enantiopure (2R,3R)-butanediol
复制标题
DOI：
10.1016/j.ymben.2014.02.003
复制
发表时间：
2014-05-01
影响因子：
8.4
通讯作者：
Zhao, Huimin
中科院分区：
文献类型：
--
作者：
Lian, Jiazhang;Chao, Ran;Zhao, Huimin
2，3-丁二醇(BDO)是一种重要的化工产品，具有广泛的工业应用前景，可由多种细菌高水平自然合成。然而，这些本土生产者的致病性是大规模生产的主要障碍。在这里，我们报告了一种工业友好的宿主，酿酒酵母，以高滴度和高产量生产BDO的工程。通过丙酮酸脱羧酶(PDCs)的失活、MTH1的过表达和适应性进化，得到的酵母以葡萄糖为唯一碳源生长，完全消除了乙醇的产生。此外，PDC-菌株同时消耗葡萄糖和半乳糖，据我们所知，这在酿酒酵母中是前所未有的。随后BDO生物合成途径的引入导致了对映体(2R，3R)-丁二醇(R-BDO)的产生。BDO生物合成途径包括胞浆乙酰乳酸合成酶(Cell ILV2)、枯草杆菌乙酰乳酸氯羧酶(BsAlsD)和内源性丁二醇脱氢酶(BDH1)。在摇瓶发酵中，产率达到理论值的70%以上。以红藻中的两种主要碳水化合物--葡萄糖和半乳糖为原料，采用分批补料发酵，合成了100g/L以上的R-BDO(1100 MM)。生产的对映体R-BDO的高效价和产量以及发酵葡萄糖和半乳糖的能力使我们的工程酵母菌株成为利用可再生资源经济高效地生产基于BDO的BDO的优良宿主。(C)2014年国际代谢工程学会。爱思唯尔公司出版，版权所有。
2,3-Butanediol (BDO) is an important chemical with broad industrial applications and can be naturally produced by many bacteria at high levels. However, the pathogenicity of these native producers is a major obstacle for large scale production. Here we report the engineering of an industrially friendly host, Saccharomyces cerevisiae, to produce BDO at high titer and yield. By inactivation of pyruvate decarboxylases (PDCs) followed by overexpression of MTH1 and adaptive evolution, the resultant yeast grew on glucose as the sole carbon source with ethanol production completely eliminated. Moreover, the pdc- strain consumed glucose and galactose simultaneously, which to our knowledge is unprecedented in S. cerevisiae strains. Subsequent introduction of a BDO biosynthetic pathway consisting of the cytosolic acetolactate synthase (cytoILV2), Bacillus subtilis acetolactate clecarboxylase (BsAlsD), and the endogenous butanediol dehydrogenase (BDH1) resulted in the production of enantiopure (2R,3R)-butanediol (R-BDO). In shake flask fermentation, a yield over 70% of the theoretical value was achieved. Using fed-batch fermentation, more than 100 g/L R-BDO (1100 mM) was synthesized from a mixture of glucose and galactose, two major carbohydrate components in red algae. The high titer and yield of the enantiopure R-BDO produced as well as the ability to en ferment glucose and galactose make our engineered yeast strain a superior host for cost-effective production of bin based BDO from renewable resources. (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.