Enzymatic semisynthesis of human insulin: an update.
Peptide bond formation can be enzymatically catalysed by the reverse reaction of proteases. Application is seen in the industrial production of human insulin. Human insulin derivative can be enzymatically prepared using either porcine insulin or the single chain B(1-29)-A(1-21) insulin precursor as the starting material. This is accomplished by either (1) digesting the starting material at Lys29 with Achromobacter lyticus protease I (Ach) and then coupling with Thr-X (X = blocking residue) (two-step reaction) or (2) subjecting Ala-B30 of porcine insulin or Gly-A1 of the single chain insulin precursor to transpeptidation with Thr-X (one-step reaction). Trypsin and Ach can be used for either type of reaction and, in the immobilized form, for the two-step reaction. Since the single chain insulin precursor can be produced by gene technology (yeast), use of immobilized trypsin or Ach and the two-step reaction using the single chain insulin precursor as the starting material ensures the continuous production of human insulin making it a feasible method for industrial manufacture.
Morihara
K
K
Department of Enzymology, University of East Asia, Yamaguchi, Japan.
D000595:Amino Acid Sequence; D000818:Animals; D004800:Enzymes, Immobilized; D006801:Humans; D007328:Insulin / Q000138:chemical synthesis*; D008969:Molecular Sequence Data; D013552:Swine
D004800:Enzymes, Immobilized; D007328:Insulin