Preparation method of recombinant I-type human collagen

Abstract

The invention discloses a method for preparing recombinant I-type human collagen. The method comprises the following steps: constructing genetically engineered bacteria for expressing proline hydroxylase, full-length COL1A1 protein and full-length COL1A2 protein; culturing the genetically engineered bacteria to obtain the recombinant I-type human collagen; wherein the amino acid sequence of the full-length COL1A1 is as shown in SEQ ID NO: 1, the amino acid sequence of the full-length COL1A2 is as shown in SEQ ID NO: 2, and the proline hydroxylase comprises P4Ha2, P4Hb and vP4H; the ratio of the copy number of the P4Ha2 to the copy number of the P4Hb to the copy number of the vP4H to the copy number of the full-length COL1A1 to the copy number of the full-length COL1A2 is (1.80 to 2.20) to (1.80 to 2.20) to (0.80 to 1.50) to (1.80 to 2.20) to 1. The method can be used for large-scale production, the purity is high, and the production cost can be reduced; the prepared recombinant I-type human collagen is completely consistent with the full-length sequence of natural I-type human collagen, the hydroxyproline content is basically consistent with that of the natural I-type human collagen, and the recombinant I-type human collagen has a triple-helix structure, has the biological activity of the natural I-type human collagen, and is free of immunogenicity and virus hidden danger.