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KMID : 0545120190290020268
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 2 p.268 ~ p.273
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Altering UDP-glucose Donor Substrate Specificity of Bacillus licheniformis Glycosyltransferase towards TDP-glucose
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Cho Kye-Woon
Kim Tae-Su
Le Tuoi Thi
Nguyen Hue Thi
Oh So-Yeong
Pandey Ramesh Prasad
Sohng Jae-Kyung
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Abstract
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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.
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KEYWORD
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Glycosyltransferase, protein engineering, enzyme Kinetics, mutation, TDP-glucose substrate specificity
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