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Efficient synthesis of l-tert-leucine through reductive amination using leucine dehydrogenase and formate dehydrogenase coexpressed in recombinant E. coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
3128 151 2014 6 PDF Available
Title
Efficient synthesis of l-tert-leucine through reductive amination using leucine dehydrogenase and formate dehydrogenase coexpressed in recombinant E. coli
Abstract

•l-tert-Leucine was synthesized through reductive amination of trimethylpyruvate.•LeuDH was coexpressed with FDH in recombinant Escherichia coli for NADH regeneration.•Substrate feeding strategy was adopted to improve the process efficiency.•1.5 M of trimethylpyruvate was stoichiometrically converted to l-Tle with >99% ee.•The continuous procedure reached a higher space-time yield of 786 g L−1 d−1.

Enantiopure l-tert-leucine (l-Tle) was synthesized through reductive amination of trimethylpyruvate catalyzed by cell-free extracts of recombinant Escherichia coli coexpressing leucine dehydrogenase (LeuDH) and formate dehydrogenase (FDH). The leudh gene from Lysinibacillus sphaericus CGMCC 1.1677 encoding LeuDH was cloned and coexpressed with NAD+-dependent FDH from Candida boidinii for NADH regeneration. The batch reaction conditions for the synthesis of l-Tle were systematically optimized. Two substrate feeding modes (intermittent and continuous) were addressed to alleviate substrate inhibition and thus improve the space-time yield. The continuous feeding process was conveniently performed in water at an overall substrate concentration up to 1.5 M, with both conversion and ee of >99% and space-time yield of 786 g L−1 d−1, respectively. Furthermore, the preparation was successfully scaled up to a 1 L scale, demonstrating the developed procedure showed a great industrial potential for the production of enantiopure l-Tle.

Graphical abstractAn efficient preparation of l-Tle was developed through reductive amination of trimethylpyruvate catalyzed by cell-free extracts of recombinant E. coli coexpressing leucine dehydrogenase and NAD+-dependent formate dehydrogenase.Figure optionsDownload full-size imageDownload as PowerPoint slide

Keywords
l-tert-Leucine; Reductive amination; Biocatalysis; Biotransformation; Bioprocess design; Optimization
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Efficient synthesis of l-tert-leucine through reductive amination using leucine dehydrogenase and formate dehydrogenase coexpressed in recombinant E. coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 91, 15 October 2014, Pages 204–209
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