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Increased productivity of l-2-aminobutyric acid and total turnover number of NAD+/NADH in a one-pot system through enhanced thermostability of l-threonine deaminase
- Source :
- Biotechnology Letters. 40:1551-1559
- Publication Year :
- 2018
- Publisher :
- Springer Science and Business Media LLC, 2018.
-
Abstract
- To strengthen NADH regeneration in the biosynthesis of l-2-aminobutyric acid (l-ABA). l-Threonine deaminase (l-TD) from Escherichia coli K12 was modified by directed evolution and rational design to improve its endurance to heat treatment. The half-life of mutant G323D/F510L/T344A at 42 °C increased from 10 to 210 min, a 20-fold increase compared to the wild-type l-TD, and the temperature at which the activity of the enzyme decreased by 50% in 15 min increased from 39 to 53 °C. The mutant together with thermostable l-leucine dehydrogenase from Bacillus sphaericus DSM730 and formate dehydrogenase from Candida boidinii constituted a one-pot system for l-ABA biosynthesis. Employing preheat treatment in the one-pot system, the biosynthesis of l-ABA and total turnover number of NAD+/NADH were 0.993 M and 16,469, in contrast to 0.635 M and 10,531 with wild-type l-TD, respectively. By using the engineered l-TD during endured preheat treatment, the one-pot system has achieved a higher productivity of l-ABA and total turnover number of coenzyme.
- Subjects :
- 0106 biological sciences
0301 basic medicine
NADH regeneration
Bioengineering
Dehydrogenase
Formate dehydrogenase
01 natural sciences
Applied Microbiology and Biotechnology
Cofactor
03 medical and health sciences
chemistry.chemical_compound
Biosynthesis
010608 biotechnology
chemistry.chemical_classification
biology
fungi
food and beverages
General Medicine
Turnover number
030104 developmental biology
Enzyme
Biochemistry
chemistry
biology.protein
NAD+ kinase
Biotechnology
Subjects
Details
- ISSN :
- 15736776 and 01415492
- Volume :
- 40
- Database :
- OpenAIRE
- Journal :
- Biotechnology Letters
- Accession number :
- edsair.doi...........1f56e3ede5705df8fed571a53cebc47e