Back to Search
Start Over
Biosynthetic L-tert-leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase
- Source :
- Electronic Journal of Biotechnology, Vol 47, Iss, Pp 83-88 (2020)
- Publication Year :
- 2020
- Publisher :
- Elsevier, 2020.
-
Abstract
- 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
- Subjects :
- 0106 biological sciences
0301 basic medicine
Chiral intermediate
lcsh:Biotechnology
Leucine dehydrogenase
medicine.disease_cause
Formate dehydrogenase
01 natural sciences
Applied Microbiology and Biotechnology
Reductive amination
03 medical and health sciences
chemistry.chemical_compound
Biosynthesis
010608 biotechnology
lcsh:TP248.13-248.65
medicine
L-tert-leucine
Escherichia coli
lcsh:QH301-705.5
Thermostability
chemistry.chemical_classification
Thermophile
Laceyella sacchari
030104 developmental biology
Enzyme
chemistry
Biochemistry
lcsh:Biology (General)
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 07173458
- Volume :
- 47
- Database :
- OpenAIRE
- Journal :
- Electronic Journal of Biotechnology
- Accession number :
- edsair.doi.dedup.....958d2a92525da45cc9c3a374758256fa