微生物来源的α-葡萄糖苷酶抑制剂高通量筛选模型的建立和初步应用

Development and Preliminary Application of High Throughput Screening Model of α-Glycosidase Inhibitor from Microorganism

本研究用昆明种小鼠十二指肠上段提取的α-葡萄糖苷酶,以4-硝基苯-α-D-吡喃葡萄糖苷为反应底物,建立α-葡萄糖苷酶抑制剂高通量筛选模型。用该模型对1276株放线菌和132株细菌等共计2224个发酵液粗提物样品进行筛选。初筛得到抑制率超过50%的阳性样品23个,初筛阳性率为1.03%。将阳性菌株再次进行发酵提取,得到新发酵液的阳性样品数为17,复筛阳性率为0.74%。其中细菌3株,放线菌13株。3个阳性样品经α-葡萄糖苷酶验证后的抑制率均在75%以上。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,可用于新型糖尿病药物的开发。

The α-glycosidase was extracted from the upper duodenum of Kunming mice. 4-Nitrophenyl-α-D-glucopyrano- side was used as substrate to establish a high throughput drug screening model of α-glycosidase inhibitor. A total of 2224 microbial fermentation extracts, including 1276 strains of actinomycetes and 132 strains of bacteria, were screened. 23 positive samples were obtained,with a 1.03% positive rate at the first screening. The 23 strains were fermented and ex- tracted for a second time, 17 positive samples were obtained, with 3 bacterium and 13 actinomycetes. The positive rate was 0.74%. 3 Positive samples were verified by ct-glycosidase to have inhibitory effects higher than 75%. In conclusion, the developed α-glycosidase inhibitor high-throughput screening modal had high practical value for developing new hypo- glycemic drugs.