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

Paper ID Volume ID Publish Year Pages File Format Full-Text
2733 127 2016 8 PDF Available
Title
A high-throughput screening method for identifying lycopene-overproducing E. coli strain based on an antioxidant capacity assay
Abstract

•24 plasmids with different orders of crt genes and SD + AS regions were constructed.•A high-throughput screening method was based on antioxidant capacity of lycopene.•There is a correlation between antioxidant capacity and lycopene content.•High lycopene production and productivity were got under the optimized conditions.•A lycopene content of 1288 mg/L and a productivity of 53.7 mg/L/h were obtained.

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 24 h.

Keywords
AOA, antioxidant capacity; ROS, reactive oxygen species; DPPH, 2,2-diphenyl-1-picrylhydrazyl; ABTS, 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) diammonium salt; FRAP, ferric reducing antioxidant power; OH, hydroxyl free radical; SD, Shine-Dalga
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A high-throughput screening method for identifying lycopene-overproducing E. coli strain based on an antioxidant capacity assay
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 112, 15 August 2016, Pages 277–284
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us