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A high-throughput screening method for identifying lycopene-overproducing E. coli strain based on an antioxidant capacity assay

文献类型： 外文期刊

作者： Xu, Xian 1 ; Jin, Weiyue 1 ; Jiang, Ling 2 ; Xu, Qing 1 ; Li, Shuang 1 ; Zhang, Zhidong 3 ; Huang, He 1 ;

作者机构： 1.Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, State Key Lab Mat Oriented Chem Engn, Nanjing 211800, Jiangsu, Peoples R China

2.Nanjing Tech Univ, Coll Food Sci & Light Ind, Nanjing 211800, Jiangsu, Peoples R China

3.Xinjiang Acad Agr Sci, Inst Microbiol, Urumqi 830091, Xinjiang Uigur, Peoples R China

4.Nanjing Tech Univ, Sch Pharmaceut Sci, 30 South Puzhu Rd, Nanjing 211800, Jiangsu, Peoples R China

关键词： Biosynthesis;Recombinant DNA;Fermentation;Optimization;High-throughput screening;Lycopene

期刊名称：BIOCHEMICAL ENGINEERING JOURNAL （ 影响因子：3.978； 五年影响因子：4.084 ）

ISSN：

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收录情况： SCI

摘要： A straightforward, high-throughput screening method for identifying recombinant Escherichia coli strains that yield high levels of lycopene was developed based on the antioxidative properties of lycopene. Twenty-four recombinant E. coli strains were constructed using plasmids with various orderings of lycopene biosynthesis genes, Shine-Dalgarno regions, and aligned spacing regions. The high-throughput screening was based on antioxidant capacity measurements; we assessed the 2,2-diphenyl-1-picrylhydrazyl method, reducing activity method, and hydroxyl free radical scavenging method; the hydroxyl free radical scavenging method was determined to be the most suitable for use with our lycopene-overproducing system. The accuracy of this method was comparable to that of assay by high-performance liquid chromatography. The strain E. coli IEB11, which had the strongest antioxidant capacity and the highest lycopene-accumulating capacity, was selected and investigated for its ability to produce lycopene at different temperatures, glycerol concentrations, and fermentation times. In the optimal conditions in a 5-L fermenter, lycopene content of 1288 mg/L and a mean productivity of 53.7 mg/L/h were obtained after 24h. (c) 2016 Elsevier B.V. All rights reserved.

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