人胰腺α-淀粉酶抑制剂高通量筛选模型的建立及其应用

High-throughput screening of human pancreatic α-amylase inhibitors

摘要:

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

Abstract:

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 16 S 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．