Thermostable peroxidase-polylysine films for biocatalysis at 90 degrees C.
Cross-linked films of poly(l-lysine) (PLL) and enzymes covalently linked to surfaces provided remarkable thermostability, enabling biocatalysis at 90 degrees C. Soret spectra, circular dichroism, and voltammetry showed that PLL films containing peroxidases or myoglobin were stable for up to 9 h at 90 degrees C, while the same enzymes in solution denatured completely within 20 min. Biocatalytic reduction of t-BuOOH with enzyme-PLL films, using rotating disk voltammetry, provided Michaelis kcat/Km values. Results showed that horseradish peroxidase (HRP)-PLL is 3-fold more active than soybean peroxidase (SBP)-PLL at 25 degrees C, but SBP-PLL is slightly more active at 90 degrees C. SBP-PLL films had 8-fold larger kcat/Km values at 90 degrees C compared to 25 degrees C. Oxidation of o-methoxyphenol to 3,3'-dimethoxy-4,4'-biphenoquinone by peroxidase-PLL-coated silica colloids gave better yields at 90 degrees C than 25 degrees C, suggesting increasing catalytic efficiency and selectivity at the higher temperature. These biocolloids were reusable with little loss of activity at 90 degrees C.
Guto
Peterson M
PM
Department of Chemistry and Institute of Materials Science, 55 North Eagleville Road, University of Connecticut, Storrs, Connecticut 06269, USA.
Kumar
Challa V
CV
Rusling
James F
JF
D002384:Catalysis; D002942:Circular Dichroism; D006139:Guaiacol / Q000737:chemistry; D006358:Hot Temperature; D008567:Membranes, Artificial; D008956:Models, Chemical; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry*; D013499:Surface Properties; D020122:tert-Butylhydroperoxide / Q000737:chemistry
D008567:Membranes, Artificial; D011107:Polylysine; D006139:Guaiacol; D020122:tert-Butylhydroperoxide; D010544:Peroxidases