CHEMICAL PROTEIN CONTAINING AN INTRAMOLECULAR CHAPERONE TYPE SEQUENCE AND ITS APPLICATION TO INSULIN PRODUCTION.

摘要

A chimeric protein comprising, from the N-terminus to the C-terminus: a) a first peptidyl fragment consisting of an amino acid sequence that has at least 60% identity with a human growth hormone domain containing at least first 20 N-terminal amino acids of SEQ ID NO: 2 in which the percent identity is determined on an amino acid sequence SEQ ID NO: 2 of the same length as the first peptidyl fragment; b) an Arg moiety, or an Lys moiety, or a second peptidyl fragment consisting of at least 2 amino acids in whose peptidyl fragment the most C-terminal acid moiety is an Arg or Lys moiety; and c) a third peptidyl fragment consisting of a human insulin precursor peptidyl fragment comprising chain A and human insulin B chain, and wherein chain A and chain B are separated by a removable peptidyl moiety of a length between 2 and 34 remains; and wherein the first peptidyl fragment is capable of increasing the bioactive conformation performance of the insulin precursor formed by contacting the recombinant protein with an auxiliary chaotropic agent compared to the bioactive conformation performance of the insulin precursor formed by putting contact the same recombinant protein that lacks the first peptidyl fragment with the chaotropic agent.