Enhanced synthesis of Coenzyme Q10 by reducing the competitive production of carotenoids in Rhodobacter sphaeroides

被引:31
作者
Zhu, Yongqiang [1 ]
Lu, Wenqiang [1 ]
Ye, Lidan [1 ]
Chen, Zhaofeng [2 ]
Hu, Weijiang [2 ]
Wang, Chenfei [2 ]
Chen, Jianbo [2 ]
Yu, Hongwei [1 ]
机构
[1] Zhejiang Univ, Inst Bioengn, Dept Chem & Biol Engn, Hangzhou 310027, Zhejiang, Peoples R China
[2] Zhejiang NHU Co Ltd, Grp Bioengn, Shaoxing 312521, Peoples R China
关键词
Coenzyme Q(10); Carotenoids; Geranylgeranyl diphosphate; ppsR; Rhodobacter sphaeroides; RECOMBINANT ESCHERICHIA-COLI; MEVALONATE PATHWAY; GENE-EXPRESSION; IN-VITRO; SYNTHASE; COQ(10); LIGHT; OVEREXPRESSION; IDENTIFICATION; BIOSYNTHESIS;
D O I
10.1016/j.bej.2017.03.019
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In Rhodobacter sphaeroides, synthesis of coenzyme Qio (CoQ(10)) shares the same precursor geranylgeranyl diphosphate (GGPP) with carotenoids. Therefore, suppression of carotenoids synthesis is supposed to pose positive effects on accumulation of CoQ(10). In this paper, the carotenogenesis pathway was repressed using two different strategies, deletion of carotenogenic genes and overexpression of ppsR, a transcriptional regulator for photosynthesis genes, respectively. Knockout of carotenoids synthesis (crt) genes resulted in undetectable carotenoids as expected. However, the production of CoQ(10), and biomass were both decreased to half as compared to the wild-type strain. In contrast, upon overexpression of ppsR, the production of carotenoids was decreased from 15.7 mg/L to 2.2 mg/L, and the CoQ(10) production and content were enhanced by 28% and 34.2%, respectively. To further enhance the production of CoQ(10), crtE was constitutively co-overexpressed with ppsR to improve the supply of GGPP as a key precursor for the isoprenoid side chain of CoQ(10). The CoQ(10) production and content of the resulting strain RspPE were increased to 73.2 mg/L and 5.67 mg/g, respectively, representing 47% and 55% improvement as compared to the wild type. This result demonstrated that appropriate reduction of carotenoids rather than complete blocking could enhance the CoQ(10) production in Rhodobacter. sphaeroides. (C) 2017 Elsevier B.V. All rights reserved.
引用
收藏
页码:50 / 55
页数:6
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