α-葡萄糖苷酶抑制剂高通量筛选模型的建立及其应用  

Establishment and application of a high-throughput model for screening α-glucosidase inhibitors

作　　者：孟鹏[1] 齐西珍[1] 郑芳[1] 任丽梅[1] 白芳[1] 白钢[1,2] 

机构地区：[1]南开大学药学院,天津300071 [2]南开大学生命科学学院,天津300071

出　　处：《微生物学报》2010年第8期1080-1086,共7页Acta Microbiologica Sinica

基　　金：国家"863计划"(2006AA020502)~~

摘　　要：【目的】针对人α-麦芽糖苷酶这个糖代谢途径中重要的靶蛋白,建立α-糖苷酶抑制剂高通量筛选模型。【方法】采用毕赤酵母表达系统克隆和表达人α-麦芽糖苷酶。利用酶的催化特性建立α-糖苷酶抑制剂筛选模型。应用该模型对放线菌代谢产物库进行高通量筛选。通过构建16SrRNA系统发育树分析阳性菌株的分类地位。【结果】首次成功克隆、表达了具催化活性的人α-麦芽糖苷酶N端结构域。针对人α-麦芽糖苷酶N端催化结构域,建立α-糖苷酶抑制剂的筛选模型。对包含近2000株放线菌代谢产物的天然产物库进行高通量筛选,最终得到20株α-麦芽糖苷酶抑制剂生产菌株。其中19株放线菌为链霉菌属,且在分类学上具有丰富的多样性。【结论】本研究建立的α-糖苷酶抑制剂高通量筛选模型具有很强的实用价值,可用于新型糖苷酶抑制剂类降糖药物的开发。[Objective] Targeted at the important enzyme in human glucose metabolic pathway,the purpose of this paper is to establish α-glucosidase inhibitors high throughput screening model.[Methods]Pichia pastoris expression system was used to clone and express the human α-maltase glucosidase.Using the catalytic properties of enzyme to establish αglucosidase inhibitor screening model.This model was applied in screening of actinomycete metabolites library.The taxonomic status of positive strains were analyzed by constructing 16S rRNA phylogenetic tree.[Results]The N-terminal catalytic domain of human α-maltase glucosidase was successfully cloned and expressed for the first time.The highthroughput screening model of α-glucosidase inhibitors was established.A natural product library containing metabolites from nearly 2000 actinomycetes was screened,20 α-maltase glucosidase inhibitor producing strains were obtained finally,of which,19 strains initially identified as Streptomyces,and showed taxonomically rich diversity.[Conclusion]The αglucosidase inhibitor high-throughput screening model has high practical value,this work laid the foundation for developing new hypoglycemic drugs.

关 键 词：Α-糖苷酶抑制剂 毕赤酵母表达系统 α-麦芽糖苷酶 放线菌 高通量药筛 

分 类 号：R927[医药卫生—药学]