改变糖基转移酶对胸苷二磷酸葡萄糖（TDP - 葡萄糖）的尿苷二磷酸葡萄糖（UDP - 葡萄糖）供体底物特异性。
Altering UDP-glucose Donor Substrate Specificity of Glycosyltransferase towards TDP-glucose.
作者信息
Cho Kye Woon, Kim Tae-Su, Le Tuoi Thi, Nguyen Hue Thi, Oh Yeong So, Pandey Ramesh Prasad, Sohng Jae Kyung
机构信息
Department of Life Science and Biochemical Engineering, Sun Moon University, Chungnam 31460, Republic of Korea.
Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University Chungnam 31460, Republic of Korea.
出版信息
J Microbiol Biotechnol. 2019 Feb 28;29(2):268-273. doi: 10.4014/jmb.1811.11009.
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 -alkyl interaction with Arg-282 and His-14, and - interaction with His and thymine ring. YjiC (H14A) variant lost its glucosylation activity with TDP-glucose validating significance of His-14 in binding of TDP-sugars.
通过定点诱变，地衣芽孢杆菌尿苷二磷酸（UDP）糖基转移酶YjiC对胸苷二磷酸（TDP）-糖的特异性得以提高。通过用色氨酸替代，对YjiC的精氨酸-282进行了鉴定和研究。比较了YjiC及其变体与几种受体底物（如7-羟基黄酮（7-HF）、4',7-二羟基异黄酮、7,8-二羟基黄酮和姜黄素）之间的转化率和动力学参数。TDP-葡萄糖和7-HF与YjiC模型的分子对接显示，与精氨酸-282和组氨酸-14存在烷基相互作用，与组氨酸和胸腺嘧啶环存在相互作用。YjiC（H14A）变体失去了对TDP-葡萄糖的糖基化活性，这证实了组氨酸-14在TDP-糖结合中的重要性。
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