Einde neue Semisynthese des Humanindulins. Tryptisch-katalysierte Transpeptidierung von Schweineinsulin mit L-Threonin-tert-butylester.

Abstract

Porcine insulin was converted to human insulin in a 1-step reaction by treatment with trypsin and threonine tert-butyl ester. The Ar (B22)-Gly (B23) bond was not cleaved by trypsin in dimethylformamide containing Tris buffer pH 5.8 after 20 h, Tris buffer pH 6.5 after 4 h or phosphate buffer pH 7.7 after 23 h. The yield of human insulin ester was 50-70%. Purification was achieved by high performance liquid chromatography or ion exchange chromatography. Upon treatment with a mixture of trifluoroacetic acid and anisol, a fully biologically active human insulin was obtained.