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Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
Expression, optimization and biological activity analysis of recombinant type XII collagen in Pichia pastoris
Qiao Gao1, Zhuo Zhang2, Rongzhan Fu1
1Engineering Research Center of Western Resource Innovation Medicine Green Manufacturing, Ministry of Education, School of Chemical Engineering, Northwest University, Xi'an 710069, China; Shaanxi R&D Center of Biomaterials and Fermentation Engineering, School of Chemical Engineering and Biotech. & Biomed. Research Institute, Northwest University, Xi'an 710069, China; Xi'an Synthetic Biology Technology and Biomaterials International Science and Technology Cooperation Base, School of Chemical Engineering, Northwest University, Xi'an 710127, China.
Abstract:
Collagen XII (COL12A1) is a type of FACIT collagen that plays an important role in the extracellular matrix structuring, participating in the regulation of collagen fiber size, and serves as a link between different components of the extracellular matrix. However, it is still unclear whether exogenous administration of collagen XII has a direct regulatory effect. In this study, we successfully produced recombinant human XII-type collagen (rh12C) through genetic engineering approach, which is composed of different functional domains. A Pichia pastoris host cell strain was constructed based on the intracellular translation regulatory mechanism of collagen, achieving a maximum yield of 4.89 g/L. After purification and structural characterization of the protein, its potential biological efficacy was evaluated through in vitro cell experiments.
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