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A new procedure for enzymatic semisynthesis of human insulin by hydrolysis of single‐chain des‐(b‐30)‐lnsulin precursor with lysyl endopeptidase
Author(s) -
Morihara Kazuyuki,
Ueno Yoshihiko
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260370712
Subject(s) - endopeptidase , chemistry , biochemistry , trypsin , hydrolysis , semisynthesis , lysine , threonine , insulin , enzyme , amino acid , biology , serine , endocrinology
It has been shown that the single‐chain des‐(B‐30)‐insulin precursor (SCI) can be converted into human insulin ester by transpeptidation using trypsin in the presence of a threonine derivative. The present study demonstrates that Achromobacter lyticus protease 1 (lysyl endopeptidase) can catalyze the transpeptidation reaction more efficiently than can trypsin. It is also shown that des‐(B‐30)‐insulin (DAI) can be produced by hydrolysis of SCI with the lysyl endopeptidase. Since it is well known that SCI can be produced by gene technology, the following method is recommended for industrial production of human insulin ester: hydrolysis of SCI with lysyl endopeptidase followed by coupling of the resulting DAI with a threonine derivative using trypsin or lysyl endopeptidase.