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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-09-01
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| Series: | Electronic Journal of Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345820300324 |
| _version_ | 1818958405552832512 |
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| author | Longxing Wang Wenjun Zhu Zhen Gao Hua Zhou Fei Cao Min Jiang Yan Li Honghua Jia Ping Wei |
| author_facet | Longxing Wang Wenjun Zhu Zhen Gao Hua Zhou Fei Cao Min Jiang Yan Li Honghua Jia Ping Wei |
| author_sort | Longxing Wang |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-20T11:25:13Z |
| format | Article |
| id | doaj.art-e6fc8a5a2a4e43a981d31937fe944260 |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-20T11:25:13Z |
| publishDate | 2020-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-e6fc8a5a2a4e43a981d31937fe9442602022-12-21T19:42:23ZengElsevierElectronic 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 |
| 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 |
| title | 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_short | 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 |
| 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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