Positional assembly of temperature-responsive biopolymers

Incorporation of enzymic cascade reactions in a microchannel for org. synthesis can be achieved if spatially controlled immobilization of a series of enzymes is possible. This project attempts positional assembly of enzymes using the inverse phase transition temp. (Tt) of elastin-like proteins (ELPs). Increasing the temp. results in aggregation and assembly of ELPs, which can be very precisely controlled. Protein engineering is used as a method to produce ELPs and their fusion proteins. These fusion proteins can be assembled onto surfaces modified with the complementary ELP, when the surface is heated above Tt. Two elastin variants were constructed that each showed the expected inverse transition behavior. Fusions of these ELPs with fluorescent proteins generated proteins with inverse transition behavior that could be controlled using varying concns. of NaCl. We are currently investigating the possibilities to functionalize microchannels with ELPs to enable positional assembly of proteins.