科研之友
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摘要
Human-like collagen (HLC) is industrially produced by using recombinant Escherichia coli and a high-cell-density fermentation technique. Similar to the collagen extracted from animal tissue, industrially produced HLC is difficult to purify because of its self-assembling property, whereby it forms aggregates of different sizes. In this study, we report the development of a purification process based on this property of HLC. The interactions of HLC fibrils were analyzed under different conditions. HLC was immobilized on Sepharose Cl-6B (HLC-Sepharose) by covalent cross-linking. This immobilization triggered lateral molecular packing of free HLC in a specific manner in a solution. We performed a purification process based on hydrogen bonding and conducted microscopic and electrophoretic analyses to assess the results of the process. We eventually obtained an electrophoretically pure product. The results show that hydrogen bonding is the main driving force that promotes the aggregation of HLC monomers, while hydrophobic bonding facilitates the stabilization of HLC aggregates. This "self-purification" method is a feasible and simple approach.
