【摘要】 【目的】针对人胰腺α-淀粉酶这个糖代谢途径中重要的靶蛋白,建立α-淀粉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胰腺α-淀粉酶;利用酶的催化特性建立α-淀粉酶抑制剂筛选模型;应用该模型对放线菌发酵液冻干物进行高通量筛选;通过构建16S rRNA系统发育树分析阳性菌株的分类地位。【结果】成功克隆、表达了具催化活性的人胰腺α-淀粉酶;建立了α-淀粉酶抑制剂的筛选模型;对近2000株放线菌的发酵液冻干物进行高通量筛选,最终得到14株α-淀粉酶抑制剂产生菌株,且在分类学上具有丰富的菌种多样性。【结论】本研究建立的α-淀粉酶抑制剂高通量筛选模型具有很强的实用价值,可用于新型淀粉酶抑制剂类降糖药物的开发。更多还原

【Abstract】 [Objective] Targeting the important enzyme in human glucose metabolic pathway,we established a high throughput screening model for human pancreatic α-amylase inhibitors.[Methods] Pichia pastoris expression system was used to clone and express the human pancreatic α-amylase;we established the α-amylase inhibitor screening model using the catalytic properties of enzyme;this model was applied in screening of actinomycete’ metabolites;the taxonomic status of positive strains were analyzed by constructing 16S rRNA phylogenetic tree.[Results] We cloned and expressed the intact gene of human pancreatic α-amylase successfully;the high-throughput screening model of α-amylase inhibitors was established;nearly 2000 actinomycete’ metabolites were screened,14 α-amylase inhibitor producing strains were obtained finally,and showed taxonomically rich diversity.[Conclusion] The α-amylase inhibitor high-throughput screening model had high practical value for developing new hypoglycemic drugs.更多还原