Sharma J K, Yadav M, Singh N P, Yadav K D S
Department of Chemistry, Udai Pratap College, Varansi 221002, India.
Prikl Biokhim Mikrobiol. 2011 Sep-Oct;47(5):584-9.
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.
已证实血红密孔菌MTCC - 137在添加了含木质素天然来源的液体培养基中能进行木质素过氧化物酶的胞外分泌。在锯末存在的情况下发现木质素过氧化物酶的分泌量最大。使用超滤和DEAE - 纤维素阴离子交换色谱法从该真菌的培养滤液中纯化该酶至均一。纯化后的木质素过氧化物酶在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中呈现一条单一蛋白带，对应分子量为40 kDa。以藜芦醇和H2O2为底物时，该酶在最适pH 2.5下的K(m)、kcat和k(cat)/K(m)值分别为61 microM、2.13 s(-1)、3.5 x 10(4) M(-1) s(-1)以及71 microM、2.13 s(-1)、3.0 x 10(4) M(-1) s(-1)。该酶的最适温度为25℃。
