Real-time analysis of enzymatic surface-initiated polymerization using surface plasmon resonance (SPR).
The kinetics of enzymatic surface-initiated polymerization of PHB on gold surface has been examined by SPR and the resultant polymer layers characterized by AFM and FT-IR spectrometry. The immobilized enzyme catalyzed surface-initiated polymerization of 3HB-CoA, resulting in the formation of a polymer brush on the surface. The rate of polymer growth from the surface was monitored by SPR in real-time. Polymer growth as measured by the increase in the resonance angle showed no apparent lag phase during the polymerization reaction. SPR analysis also revealed that the thickness of the polymer film could be controlled by varying the initial enzyme density on the surface. The average thicknesses of the PHB film after polymerization reaction were 95, 45 and 15 nm for the surfaces that were treated with 0.5, 0.3 and 0.1*10(-6) M of enzyme, respectively. The binding of PHA synthase at different concentration to the mixed SAMs and subsequent polymerization.
Kim
Young-Rok
YR
Department of Food Science, 312 Stocking Hall, Cornell University, Ithaca, NY 14853, USA.
Paik
Hyun-Jong
HJ
Ober
Christopher K
CK
Coates
Geoffrey W
GW
Mark
Sonny S
SS
Ryan
Thomas E
TE
Batt
Carl A
CA
D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D020118:Cupriavidus necator / Q000201:enzymology / Q000235:genetics; D004800:Enzymes, Immobilized; D006046:Gold; D018625:Microscopy, Atomic Force; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives; D009942:Organometallic Compounds; D011091:Polyesters / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012825:Silicon; D020349:Surface Plasmon Resonance
D004800:Enzymes, Immobilized; D009942:Organometallic Compounds; D011091:Polyesters; D011994:Recombinant Proteins; C088321:nickel nitrilotriacetic acid; D006046:Gold; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase; D009571:Nitrilotriacetic Acid; D012825:Silicon