High-efficiency Secretory Expression and Characterization of the Recombinant Type III Human-like Collagen in Pichia Pastoris

Overview

Journal Bioresour Bioprocess

Specialty Biochemistry

Date 2024 Apr 22

PMID 38647563

Authors

Zhi-Xiang Xiang

Jin-Song Gong

Jin-Hao Shi

Chun-Fang Liu

Heng Li

Chang Su

Min Jiang

Zheng-Hong Xu

Jin-Song Shi

Affiliations

Soon will be listed here.

Abstract

Collagen, the highest content protein in the body, has irreplaceable biological functions, and it is widespread concerned in food, beauty, and medicine with great market demand. The gene encoding the recombinant type III human-like collagen α1 chain fragment was integrated into P. pastoris genome after partial amino acids were substituted. Combined with promoter engineering and high-density fermentation technology, soluble secretory expression with the highest yield of 1.05 g L was achieved using two-stage feeding method, and the purity could reach 96% after affinity purification. The determination of N/C-terminal protein sequence were consistent with the theoretical expectation and showed the characteristics of Gly-X-Y repeated short peptide sequence. In amino acid analysis, glycine shared 27.02% and proline 23.92%, which were in accordance with the characteristics of collagen. Ultraviolet spectrum combined with Fourier transform infrared spectroscopy as well as mass spectrometry demonstrated that the target product conformed to the characteristics of collagen spectrums and existed as homologous dimer and trimer in the broth. This work provided a sustainable and economically viable source of the recombinant type III human-like collagen.

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