Characterization of ELP-fused -Transaminase and Its Application for the Biosynthesis of -Amino Acid
- Authors
- Jeon, Hyunwoo; Sarak, Sharad; Lee, Sang-Hyuk; Bea, Han-Seop; Patil, Mahesh; Kim, Geon-Hee; Kim, Byung-Gee; Won, Jong In; Yun, Hyungdon
- Issue Date
- Sep-2018
- Publisher
- KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
- Keywords
- (S)-omega-transaminase; elastin-like polypeptides; chiral amines; bio-catalysis; thermostability
- Citation
- BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, v.23, no.5, pp.481 - 489
- Journal Title
- BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
- Volume
- 23
- Number
- 5
- Start Page
- 481
- End Page
- 489
- URI
- https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/3278
- DOI
- 10.1007/s12257-018-0268-7
- ISSN
- 1226-8372
- Abstract
- Optically pure amines, -amino acids and -amino acids are the valuable precursors to produce biologically active compounds. The -TAs are the class of enzymes which are widely used to produce such compounds. In this work (S)--transaminase from the thermophilic eubacterium Sphaerobacter thermophilus (St-TA) was fused with Elastin-like polypeptides (ELPs) through the cloning process and expressed in E. coli cells. The characterization of this fusion complex was performed with respect to thermostability and effect of DMSO. Where in case of St-TA-ELP-V-60, major difference in the transition temperature (T-t) was observed, wherein a T-t of 38 and 70 degrees C was observed at the increasing concentration of DMSO from 5 to 25% (v/v). Interestingly, these fusion proteins the activity was preserved even after the aggregation of fusion complex at Tt. The substrate specificity and product inhibition analysis showed that -TA-ELPs had comparable results as that of wild type -TA. Moreover, the fused -TA could be efficiently reused for up to 20 batches of transamination reaction. Furthermore, the applicability of the fusion protein for the production of a sitagliptin precursor (R)-3-amino-4-(2,4,5-triflurophenyl) butanoic acid (3-ATfBA) was evaluated, wherein 3-ATfBA was synthesized with good conversion (65%).
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Collections - College of Engineering > Chemical Engineering Major > 1. Journal Articles
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