MEP代谢通路组合优化提高β-胡萝卜素产量
Combinatorial Optimization of MEP Pathway to Improve β-Carotene Yield
刘乐山 1刘海元 1谢丽萍 1胡又佳1
作者信息
- 1. 中国医药工业研究总院,上海201203
- 折叠
摘要
通过简单可靠的方法提升目标产物产量是代谢工程的一大目的,但代谢流量的不均衡利用与酶活性不足往往会限制生物合成的效率.目前已有许多关于单个酶的优化研究,本研究展示了一种可以平衡代谢流量的新型组合优化方法,该方法基于Gibson组装(GA)与双顺反子设计(BCD)标准起始元件.以MEP通路为例,通过选取2个限速酶编码基因:dxs和idi,与5个绝缘BCD起始元件,在有限步骤内产生了一个组合优化库,再以β-胡萝卜素为指示物进行筛选,使产量提升至10倍以上.研究结果证实了该组合优化方法的有效性与代谢通路优化工程的巨大潜力,同时提示这种优化方法适用于一系列次级代谢通路.
Abstract
One goal of metabolic engineering is to improve yields with robust and simply executed methods.However,unbalanced utilization of metabolic pathway flux and insufficient activity of enzymes are the major challenges of microbial biosynthesis efficiency.Numerous researches on single enzyme optimization have been reported through studies in microorganisms,our work presents a new combinatorial optimization approach which balances metabolic flux with modular genetic elements.This approach is based on Gibson Assembly (GA) and bicistronic design (BCD) standard initiation elements.We used methylerythritol 4-phosphate (MEP) pathway as an example to demonstrate this approach.Two rate-limiting enzymes coding genes dxs and idi,and five insulated modular initiation elements were employed to generate a combination library in limited steps.A β-carotene indicated screening method was adopted to obtain a recombinant strain that increased the production yield to more than 10 folds.The results demonstrated the effectiveness of this combinatorial optimization approach and the potential of pathway optimization engineering.We believed that this approach could be applicable for a range of secondary metabolic pathways.
关键词
MEP代谢通路/Gibson组装/组合优化/β-胡萝卜素Key words
MEP pathway/Gibson Assembly/combinatorial optimization/β-carotene引用本文复制引用
出版年
2018
NETL
NSTL
维普
万方数据