Enzymatic surface-initiated polymerization: a novel approach for the in situ solid-phase synthesis of biocompatible polymer poly(3-hydroxybutyrate).
A novel system for surface-initiated enzymatic polymerization of a film of polyhydroxyalkanoate (PHA) on solid surfaces has been developed and characterized. PHAs are aliphatic polyesters produced by a variety of microorganisms as a reserve of carbon and energy, and their properties range from elastomers to thermoplastics, depending on their monomeric composition. The PHA synthase from Ralstonia eutropha H16 was expressed as a poly-histidine fusion in Escherichia coli and immobilized onto several solid substrates through a transition-metal complex, Ni(2+)-nitrilotriacetic acid. The immobilized PHA synthase catalyzed the surface-initiated polymerization of 3-(R)-hydroxybutyryl-CoA, forming a polymer film with a uniform thickness on the surface. In this work, we describe the patterned immobilization of the intact enzyme on silicon and subsequent enzymatic polymerization. The immobilized enzyme had a lower specific activity and did not exhibit a lag phase as compared to the soluble enzyme.
Kim
Young-Rok
YR
Department of Food Science, Cornell University, Ithaca, New York 14853, USA.
Paik
Hyun-Jong
HJ
Ober
Christopher K
CK
Coates
Geoffrey W
GW
Batt
Carl A
CA
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics; D001672:Biocompatible Materials; D020118:Cupriavidus necator / Q000201:enzymology; D004798:Enzymes / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D006885:Hydroxybutyrates / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D011091:Polyesters / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D000094122:Polyhydroxybutyrates
D000217:Acyltransferases; D001672:Biocompatible Materials; D004798:Enzymes; D006885:Hydroxybutyrates; D011091:Polyesters; D011108:Polymers; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C077225:poly(3-hydroxyalkanoic acid) synthase