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Sheng Wu Gong Cheng Xue Bao. 2019 Feb 25;35(2):319-326. doi: 10.13345/j.cjb.180266.

[Preparation and characterization of recombinant human-source collagen].

[Article in Chinese; Abstract available in Chinese from the publisher]

Author information

1
Shaanxi HuiKang Bio-Tech Co Ltd., Xi'an 710054, Shaanxi, China.

Abstract

in English, Chinese

This study aimed to obtain a recombinant human-source collagen for industrialization. First, based on the Gly-X-Y sequence of human type I collagen, we optimized the hydrophilic Gly-X-Y collagen peptide, designed the human collagen amino acid sequence and the corresponding nucleotide sequence. Next, the expression vector pPIC9K-COL was constructed via endonuclease digestion technology. We obtained an engineering strain of human-source collagen by electrotransforming Pichia pastoris, and then it was fermented, purified and identified. As a result, the expression level reached 4.5 g/L and the purity was over 95%. After amino acid N-terminal sequencing, molecular weight analysis, amino acid analysis and collagenase degradation test, we confirmed that the obtained collagen was consistent with designed primary structure of human-source collagen. After freeze-drying, we analyzed the collagen by scanning electron microscope and cell cytotoxicity, confirming that the collagen has porous fiber reticular structure and superior cytocompatibility. This indicates that human-source collagen has potential to be applied as biomedical material. In conclusion, we successfully obtained the expected human-source collagen and laid a foundation to its further application.

KEYWORDS:

Pichia pastoris; biomaterial; collagen; human-source

PMID:
30806062
DOI:
10.13345/j.cjb.180266
[Indexed for MEDLINE]
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