新颖糖基转移酶对抗菌药物金霉素的糖基化修饰  

Enzymatic glycosylation of antibiotics chlortetracycline by a new glycosyltransferase

作　　者：戴舒远 赵锡澍 解可波 王霞[1] 戴均贵[2] DAI Shu-yuan ZHAO Xi-shu XIE Ke-bo WANG Xia DAI Jun-gui(Beijing No. 35 High School, Beijing 100034, China （DAI Shu-yuan, ZHAO Xi-shu, WANG Xia Institute of Materia Medica, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Beijing 100050, China （XIE Ke-bo, DAI Jun-gui)

机构地区：[1]北京市第三十五中学,100034 [2]中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《中国医药生物技术》2017年第3期221-226,共6页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81602999)

摘　　要：目的发掘新颖糖基转移酶并对金霉素进行糖基化修饰。方法对来源于木立芦荟的重组糖基转移酶进行金霉素糖基化功能筛选,发现重组AaGT1能催化金霉素与葡萄糖糖基供体进行反应。放大反应液经乙酸乙酯萃取去除未反应底物,剩余含糖基化产物的水相部分采用反相半制备HPLC技术进行分离纯化。通过质谱和核磁共振波谱等技术对产物结构进行鉴定;对底物与糖基化产物的水溶性进行测定,并评价两者抑菌活性。结果糖基转移酶AaGT1催化金霉素获得两个金霉素异构体的糖基化产物,异金霉素10-O-β-D-葡萄糖苷和4-差向异金霉素10-O-β-D-葡萄糖苷。异金霉素10-O-β-D-葡萄糖苷的水溶性是盐酸金霉素水溶性的3.3倍;糖基化产物对金黄色葡萄球菌没有明显的抑制活性。结论糖基转移酶AaGT1对金霉素能够进行糖基化,产物异金霉素10-O-β-D-葡萄糖苷和4-差向异金霉素10-O-β-D-葡萄糖苷均为新化合物,对底物进行糖基化修饰能提高其水溶性。Objective To mine novel glycosyltransferase with the ability to glycosylate antibiotics chlortetracycline. Methods After preliminarly sreening, glycosyltransferase AaGT1 from A. arborescens was found to be capable of glycosylating chlortetracycline. AaGT1 was utilized to catalyze the glycosylation of chlortetracycline with uridine diphosphate glucose. To remove the unreacted chlortetracycline, the reaction solution was extracted with EtOAc for five times. And the rest aqueous phase was dried by removal of water under reduced pressure, then purified by semi-preparative reversed-phase HPLC. The structures of glycosylated products were determined by HRMS and NMR spectroscopic data analysis. In addition, water solubility of glycosylated product and substrate were measured, and their antibacterial activities were evaluated. Results Two glycosylated products were obtained and identified as iso-chlortetracycline 10-O-β-D-glucoside and 4-epi-iso-chlortetracycline 10-O-β-D-glucoside. The solubility of iso-chlortetracycline 10-O-β-D-glucoside in water was increased by 3.3 times compared with that of chlortetracycline hydrochloride. However, the glycosylated products showed no significant antibacterial activity against Staphylococcus aureus. Conclusion Iso-chlortetracycline 10-O-β-D-glucoside, 4-epi-iso-chlortetracycline 10-O-β-D-glucoside are novel compounds, and glycosylation of drugs may increase their water solubility.

关 键 词：金霉素 糖基化 糖基转移酶 木立芦荟 

分 类 号：R914[医药卫生—药物化学]