Abstract:In this study, based on the self-aggregation characteristics of the self-assembling ability of ELK16 in Escherichia coli, Aglycin was linked to the self-assembling peptide via an intein, Mxe GyrA, to construct a recombinant expression vector pET30a-Aglycin-Mxe-PT-ELK16 capable of self-cleavage in vitro. The recombinant plasmid was transformed into E. coli BL21 (DE3), and the protein expression conditions were optimized to obtain the target protein aggregates. Then, the DTT cleavage conditions were optimized for the aggregates. After further purification, Aglycin peptide with a purity of 98.15% and yield of 5.53 mg/g cell pellet wet weight was finally obtained. The yield of Aglycin peptide obtained by recombinant expression was almost 100 times higher than that of the currently used chemical extraction method. In addition, in vitro experiments have shown that the IC50 value (36.48 μmol/L) of Aglycin peptide for α-glucosidase inhibition was lower than that of acarbose (991.33 μmol/L), indicating that the inhibitory activity of Aglycin peptide against α-glucosidase was much stronger than that of acarbose. This further demonstrates the potential of Aglycin peptide as an α-glucosidase inhibitor.
