Fabrication of Engineered Collagen-Like Biopolymers

Collagen, the most abundant extracellular matrix protein, and collagen-like materials have been widely used in tissue scaffold and drug delivery applications. Recombinant human collagen using the native gene can be expressed in mammalian and yeast cells. However, the characteristic Gly-X-Y protein sequence repetition of the triple-helical domain adds an extra degree of complexity to the de novo gene synthesis that results in incorrect oligonuclucleotide hybridizations. This has, up until now, prevented the synthesis of full-length collagen-like biopolymers. Using a computational algorithm, we have successfully created a gene encoding native human collagen III. This gene has been designed to enable modular modifications and facilitate the fabrication of collagen-like variants. We have expressed these biopolymers in Saccharomyces cerevisiae using an integration/plasmid system, and these protein products have been purified and characterized. The strategy we have developed sets up the foundation for creating variants of collagen for tailored applications.