Expression, Purification and Characterization of the Recombinant Hirudin Variant iii in the Bacillus Subtilis

摘要

Hirudin is the most potent natural inhibitor of thrombin and a powerful anticoagulant. Large-scale production of recombinant hirudin is desirable for therapy. In this study, the gene encoding hirudin variant III was redesigned and synthesized by using Bacillus subtilis preferred codons, and the recombinant hirudin variant III (rHV3) was overexpressed in B. subtilis DB403 with strong anticoagulation activity for the first time. The hirudin activity from the supernatant of culture with optimized expression conditions could reach 210 ATU/ml. The protein in culture supernatant was precipitated by trichloroacetic acid, then desalted by ultrafiltration and purified by anion exchange chromatography. Strong anion Q F.F. performed better than weak anion DEAE F.F.