Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase

Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase

Background: L-tert-Leucine has been widely used in pharmaceutical, chemical, and other industries as a vital chiral intermediate. Compared with chemical methods, enzymatic methods to produce L-tert-leucine have unparalleled advantages. Previously, we found a novel leucine dehydrogenase from the halo...

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Main Authors: Longxing Wang, Wenjun Zhu, Zhen Gao, Hua Zhou, Fei Cao, Min Jiang, Yan Li, Honghua Jia, Ping Wei
Format: Article
Language:English
Published: Elsevier 2020-09-01
Series:Electronic Journal of Biotechnology
Subjects:
Chiral intermediate
Formate dehydrogenase
Laceyella sacchari
Leucine dehydrogenase
L-tert-leucine
Reductive amination
Online Access:http://www.sciencedirect.com/science/article/pii/S0717345820300324
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spelling doaj-e6fc8a5a2a4e43a981d31937fe9442602020-11-25T03:41:09ZengElsevierElectronic Journal of Biotechnology0717-34582020-09-01478388Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenaseLongxing Wang0Wenjun Zhu1Zhen Gao2Hua Zhou3Fei Cao4Min Jiang5Yan Li6Honghua Jia7Ping Wei8College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCorresponding authors.; College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCorresponding authors.; College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, ChinaBackground: L-tert-Leucine has been widely used in pharmaceutical, chemical, and other industries as a vital chiral intermediate. Compared with chemical methods, enzymatic methods to produce L-tert-leucine have unparalleled advantages. Previously, we found a novel leucine dehydrogenase from the halophilic thermophile Laceyella sacchari (LsLeuDH) that showed good thermostability and great potential for the synthesis of L-tert-leucine in the preliminary study. Hence, we manage to use the LsLeuDH coupling with a formate dehydrogenase from Candida boidinii (CbFDH) in the biosynthesis of L-tert-leucine through reductive amination in the present study. Result: The double-plasmid recombinant strain exhibited higher conversion than the single-plasmid recombinant strain when resting cells cultivated in shake flask for 22 h were used. Under the optimized conditions, the double-plasmid recombinant E. coli BL21 (pETDute-FDH-LDH, pACYCDute-FDH) transformed 1 mol·L-1 trimethylpyruvate (TMP) completely into L-tert-leucine with greater than 99.9% ee within 8 h. Conclusions: The LsLeuDH showed great ability to biosynthesize L-tert-leucine. In addition, it provided a new option for the biosynthesis of L-tert-leucine. How to cite: Wang L, Zhu W, Gao Z, et al. Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase. Electron J Biotechnol 2020;47. https://doi.org/10.1016/j.ejbt.2020.07.001http://www.sciencedirect.com/science/article/pii/S0717345820300324Chiral intermediateFormate dehydrogenaseLaceyella sacchariLeucine dehydrogenaseL-tert-leucineReductive amination
collection DOAJ
language English
format Article
sources DOAJ
author Longxing Wang
Wenjun Zhu
Zhen Gao
Hua Zhou
Fei Cao
Min Jiang
Yan Li
Honghua Jia
Ping Wei
spellingShingle Longxing Wang
Wenjun Zhu
Zhen Gao
Hua Zhou
Fei Cao
Min Jiang
Yan Li
Honghua Jia
Ping Wei
Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
Electronic Journal of Biotechnology
Chiral intermediate
Formate dehydrogenase
Laceyella sacchari
Leucine dehydrogenase
L-tert-leucine
Reductive amination
author_facet Longxing Wang
Wenjun Zhu
Zhen Gao
Hua Zhou
Fei Cao
Min Jiang
Yan Li
Honghua Jia
Ping Wei
author_sort Longxing Wang
title Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
title_short Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
title_full Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
title_fullStr Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
title_full_unstemmed Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
title_sort biosynthetic l-tert-leucine using escherichia coli co-expressing a novel nadh-dependent leucine dehydrogenase and a formate dehydrogenase
publisher Elsevier
series Electronic Journal of Biotechnology
issn 0717-3458
publishDate 2020-09-01
description Background: L-tert-Leucine has been widely used in pharmaceutical, chemical, and other industries as a vital chiral intermediate. Compared with chemical methods, enzymatic methods to produce L-tert-leucine have unparalleled advantages. Previously, we found a novel leucine dehydrogenase from the halophilic thermophile Laceyella sacchari (LsLeuDH) that showed good thermostability and great potential for the synthesis of L-tert-leucine in the preliminary study. Hence, we manage to use the LsLeuDH coupling with a formate dehydrogenase from Candida boidinii (CbFDH) in the biosynthesis of L-tert-leucine through reductive amination in the present study. Result: The double-plasmid recombinant strain exhibited higher conversion than the single-plasmid recombinant strain when resting cells cultivated in shake flask for 22 h were used. Under the optimized conditions, the double-plasmid recombinant E. coli BL21 (pETDute-FDH-LDH, pACYCDute-FDH) transformed 1 mol·L-1 trimethylpyruvate (TMP) completely into L-tert-leucine with greater than 99.9% ee within 8 h. Conclusions: The LsLeuDH showed great ability to biosynthesize L-tert-leucine. In addition, it provided a new option for the biosynthesis of L-tert-leucine. How to cite: Wang L, Zhu W, Gao Z, et al. Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase. Electron J Biotechnol 2020;47. https://doi.org/10.1016/j.ejbt.2020.07.001
topic Chiral intermediate
Formate dehydrogenase
Laceyella sacchari
Leucine dehydrogenase
L-tert-leucine
Reductive amination
url http://www.sciencedirect.com/science/article/pii/S0717345820300324
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