Synthesis and characterization of a soluble, temperature-sensitive polymer-conjugated enzyme.
The enzyme, alkaline phosphatase, has been conjugated to a temperature-sensitive polymer which exhibits a lower critical solution temperature (LCST). A series of copolymers containing different molar ratios of N-isopropylacrylamide(NIPAAm) and N-acryloxysuccinimide(NAS) were synthesized and then conjugated to the enzyme. These polymer-enzyme conjugates precipitate and flocculate in aqueous solution above the LCST, and redissolve when cooled below that temperature. The kinetics of the conjugated enzymes have been characterized as a function of temperature and compared to free enzyme. The effect of the conjugation degree between polymer and enzyme on the activity of the conjugated enzymes was also investigated.
Park
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TG
Center for Bioengineering, University of Washington, Seattle 98195.
Hoffman
A S
AS
D000179:Acrylates / Q000737:chemistry*; D000180:Acrylic Resins / Q000737:chemistry*; D000469:Alkaline Phosphatase / Q000737:chemistry*; D004795:Enzyme Stability; D004952:Esters; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D012995:Solubility; D013388:Succinimides / Q000737:chemistry*; D013696:Temperature
D000179:Acrylates; D000180:Acrylic Resins; D004952:Esters; D013388:Succinimides; C052970:poly-N-isopropylacrylamide; C053911:N-succinimidyl acrylate; D000469:Alkaline Phosphatase