目的对海洋芽孢杆菌B-9987中糖基转移酶BmmGT1糖基受体的广谱性进行探究。方法以UDPD-葡萄糖为糖基供体,与不同糖基受体进行体外酶促反应,通过高效液相色谱(HPLC)检测反应产物,并采用高分辨质谱(HRMS)手段对糖基化产物进行初步鉴定。结果糖基转移酶BmmGT1能够以UDP-D-葡萄糖为糖基供体,识别两性霉素B以及氯霉素,分别生成两性霉素B双糖基化和单糖基化衍生物以及氯霉素单糖基化衍生物,得到3个糖基化产物。结论糖基转移酶BmmGT1对糖基受体的选择有灵活性,可作为潜在的工具酶用于化合物结构多样性研究。
Objective To study the substrate promiscuity of macrolide glycosyltransferase BmmGT1 from marine Bacillus regarding sugar receptor. Methods In vitro enzymatic reactions were set up using dif- ferent antibiotics as sugar receptors and UDP-D-glucose as sugar donor, which were followed by HPLC and HRMS analysis. Results BmmGT1 was able to transfer glucosyl moiety onto Amphotericin B (Am- ph B) and Chloramphenicol (Chl), leading to formation of two glucosylated Amph B derivatives and one glucosylated Chl derivative, respectively. Conclusion BmmGT1 exhibited broad substrate specificity with regards to sugar receptor, which demonstrated its potential application as an effective tool enzyme in structure diversification of natural products.