2019 ; Vol.29-2: 268~273
|Author||Kye Woon Cho, Tae-Su Kim, Tuoi Thi Le, Hue Thi Nguyen, Yeong So Oh, Ramesh Prasad Pandey, Jae Kyung Sohng|
|Place of duty||Department of Life Science and Biochemical Engineering, Sun Moon University|
|Title||Altering UDP-glucose Donor Substrate Specificity of Bacillus licheniformis Glycosyltransferase towards TDP-glucose|
J. Microbiol. Biotechnol.2019 ;
|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 His14 and thymine ring. YjiC (H14A) variant lost its glucosylation activity with TDP-glucose validating significance of His-14 in binding of TDP-sugars.|
|Key_word||Glycosyltransferase, protein engineering, enzyme Kinetics, mutation, TDP-glucose substrate specificity|
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