Purification and characterisation of lignin peroxidase from Pycnoporus sanguineus MTCC-137.
Extracellular secretion of lignin peroxidase from Pycnoporus sanguineus MTCC-137 in the liquid culture growth medium amended with lignin containing natural sources has been shown. The maximum secretion of lignin peroxidase has been found in the presence of saw dust. The enzyme has been purified to homogeneity from the culture filtrate of the fungus using ultrafiltration and anion exchange chromatography on DEAE-cellulose. The purified lignin peroxidase gave a single protein band in sodium dodecylsulphate polyacrylamide gel electrophoresis corresponding to the molecular mass 40 kDa. The K(m)(, kcat) and k(cat)/K(m) values of the enzyme using veratryl alcohol and H2O2 as the substrate were 61 microM, 2.13 s(-1), 3.5 x 10(4) M(-1) s(-1) and 71 microM, 2.13 s(-1), 3.0 x 10(4) M(-1) s(-1) respectively at the optimum pH of 2.5. The temperature optimum of the enzyme was 25 degrees C.
Department of Chemistry, Udai Pratap College, Varansi 221002, India.
J K
JK
Sharma
M
M
Yadav
N P
NP
Singh
K D S
KD
Yadav
D001592:Benzyl Alcohols / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006861:Hydrogen Peroxide / Q000737:chemistry; D007700:Kinetics; D008031:Lignin / Q000737:chemistry / Q000378:metabolism; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification*; D055494:Pycnoporus / Q000201:enzymology*; D013696:Temperature
D001592:Benzyl Alcohols; D008031:Lignin; D006861:Hydrogen Peroxide; D010544:Peroxidases; C042858:lignin peroxidase; C042197:veratryl alcohol