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http://dx.doi.org/10.4014/jmb.1811.11009

Altering UDP-Glucose Donor Substrate Specificity of Bacillus licheniformis Glycosyltransferase towards TDP-Glucose  

Cho, Kye Woon (Department of Life Science and Biochemical Engineering, Sun Moon University)
Kim, Tae-Su (Department of Life Science and Biochemical Engineering, Sun Moon University)
Le, Tuoi Thi (Department of Life Science and Biochemical Engineering, Sun Moon University)
Nguyen, Hue Thi (Department of Life Science and Biochemical Engineering, Sun Moon University)
Oh, So Yeong (Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University Chungnam)
Pandey, Ramesh Prasad (Department of Life Science and Biochemical Engineering, Sun Moon University)
Sohng, Jae Kyung (Department of Life Science and Biochemical Engineering, Sun Moon University)
Publication Information
Journal of Microbiology and Biotechnology / v.29, no.2, 2019 , pp. 268-273 More about this Journal
Abstract
The specificity of a Bacillus licheniformis uridine diphosphate (UDP) glycosyltransferase, YjiC, was increased towards thymidine diphosphate (TDP)-sugar by site-directed mutagenesis. The Arg-282 of YjiC was identified and investigated by substituting with Trp. Conversion rate and kinetic parameters were compared between YjiC and its variants with several acceptor substrates such as 7-hydroxyflavone (7-HF), 4',7-dihydroxyisoflavone, 7,8-dihydroxyflavone and curcumin. Molecular docking of TDP-glucose and 7-HF with YjiC model showed pi-alkyl interaction with Arg-282 and His-14, and pi-pi interaction with $His^{14}$ and thymine ring. YjiC (H14A) variant lost its glucosylation activity with TDP-glucose validating significance of His-14 in binding of TDP-sugars.
Keywords
Glycosyltransferase; protein engineering; enzyme kinetics; mutation; TDP-glucose substrate specificity;
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