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
第一作者: Xu, Xian
作者: Xu, Xian;Jin, Weiyue;Xu, Qing;Li, Shuang;Huang, He;Jiang, Ling;Zhang, Zhidong;Huang, He
作者机构:
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.
分类号: Q503
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