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

Abstract:

Abstract: [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 α-glucosidase 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 high-throughput 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 α-glucosidase inhibitor high-throughput screening model has high practical value, this work laid the foundation for developing new hypoglycemic drugs.