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Bioconversion of l-glutamic acid to α-ketoglutaric acid by an immobilized whole-cell biocatalyst expressing l-amino acid deaminase from Proteus mirabilis

Paper ID Volume ID Publish Year Pages File Format Full-Text
23343 43433 2014 9 PDF Available
Title
Bioconversion of l-glutamic acid to α-ketoglutaric acid by an immobilized whole-cell biocatalyst expressing l-amino acid deaminase from Proteus mirabilis
Abstract

•Developed an immobilized whole-cell biocatalytic process for a-KG production from glutamic acid.•Compared the suitability of E. coli and B. subtilis as potential biocatalysts overexpressing l-AAD.•The performance of B. subtilis was superior to that of E. coli though both strains were active.•The highest a-KG yield was 31% using 15.0 g/L glutamic acid and 20 g/L whole-cell B. subtilis.•Immobilizing whole cells with alginate increased the recyclability by 23.33% per cycle.

The goal of this work was to develop an immobilized whole-cell biocatalytic process for the environment-friendly synthesis of α-ketoglutaric acid (α-KG) from l-glutamic acid. We compared the suitability of Escherichia coli and Bacillus subtilis strains overexpressing Proteus mirabilisl-amino acid deaminase (l-AAD) as potential biocatalysts. Although both recombinant strains were biocatalytically active, the performance of B. subtilis was superior to that of E. coli. With l-glutamic acid as the substrate, α-KG production levels by membranes isolated from B. subtilis and E. coli were 55.3 ± 1.73 and 21.7 ± 0.39 μg/mg protein/min, respectively. The maximal conversion ratio of l-glutamic acid to α-KG was 31% (w/w) under the following optimal conditions: 15 g/L l-glutamic acid, 20 g/L whole-cell biocatalyst, 5 mM MgCl2, 40 °C, pH 8.0, and 24-h incubation. Immobilization of whole cells with alginate increased the recyclability by an average of 23.33% per cycle. This work established an efficient one-step biotransformation process for the production of α-KG using immobilized whole B. subtilis overexpressing P. mirabilisl-AAD. Compared with traditional multistep chemical synthesis, the biocatalytic process described here has the advantage of reducing environmental pollution and thus has great potential for the large-scale production of α-KG.

Keywords
Whole-cell biocatalyst; α-Ketoglutaric acid; Proteus mirabilis; Glutamic acid; Bacillus subtilis
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Bioconversion of l-glutamic acid to α-ketoglutaric acid by an immobilized whole-cell biocatalyst expressing l-amino acid deaminase from Proteus mirabilis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 169, 10 January 2014, Pages 112–120
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