RBS文库调控重组大肠杆菌β-胡萝卜素合成途径关键基因提高β-胡萝卜素合成能力

Modulating expression of key genes within β-carotene synthetic pathway in recombinant Escherichia coli with RBS library to improve β-carotene production

摘要:

β-胡萝卜素属于类胡萝卜素家族的一员，在药品、保健品、化妆品和食品行业有广泛的应用。本研究通过用RBS文库对重组大肠杆菌CAR005中β-胡萝卜素合成途径的关键基因dxs、idi和crt操纵子进行调控来提高β-胡萝卜素合成能力。研究发现3个基因分别用RBS文库调控后，与起始菌株相比β-胡萝卜素产量最高分别有7%、11%和17%的提高，表明使用RBS文库调控比使用多个固定强度启动子调控能筛选到更有利于目标产品合成的基因表达强度。三基因组合调控后，β-胡萝卜素产量相对于CAR005菌株提高了35%。同时发现，单基因文库筛选到的最优强度对于组合调控来说，未必是最优强度。本研究为利用基因表达调控优化目标产物合成途径提供了一种新的方案。

Abstract:

β-carotene belongs to carotenoids family, widely applied in pharmaceuticals, neutraceuticals, cosmetics and food industries. In this study, three key genes (dxs, idi, and crt operon) within β-carotene synthetic pathway in recombinant Escherichia coli strain CAR005 were modulated with RBS Library to improve β-carotene production. There were 7%, 11% and 17% increase of β-carotene yield respectively after modulating dxs, idi and crt operon genes with RBS Library, demonstrating that modulating gene expression with regulatory parts libraries would have more opportunities to obtain optimal production of target compound. Combined modulation of crt operon, dxs and idi genes led to 35% increase of β-carotene yield compared to parent strain CAR005. The optimal gene expression strength identified in single gene modulation would not be the optimal strength when used in combined modulation. Our study provides a new strategy for improving production of target compound through modulation of gene expression.