Purification and characterization of laccase from the white rot fungus Trametes versicolor.
Laccase is one of the ligninolytic enzymes of white rot fungus Trametes versicolor 951022, a strain first isolated in Korea. This laccase was purified 209-fold from culture fluid with a yield of 6.2% using ethanol precipitation, DEAE-Sepharose, Phenyl-Sepharose, and Sephadex G-100 chromatography. T. versicolor 951022 excretes a single monomeric laccase showing a high specific activity of 91,443 U/mg for 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as a substrate. The enzyme has a molecular mass of approximately 97 kDa as determined by SDS-PAGE, which is larger than those of other laccases reported. It exhibits high enzyme activity over broad pH and temperature ranges with optimum activity at pH 3.0 and a temperature of 50 degrees C. The Km value of the enzyme for substrate ABTS is 12.8 micrometer and its corresponding Vmax value is 8125.4 U/mg. The specific activity and substrate affinity of this laccase are higher than those of other white rot fungi, therefore, it may be potentially useful for industrial purposes.
Han
Mun-Jung
MJ
Division of Biological Sciences, Kangwon National University, Chuncheon, Republic of Korea.
Han
Moon-Jeong
MJ
Choi
Hyoung-Tae
HT
Song
Hong-Gyu
HG
D052160:Benzothiazoles; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005590:Fractional Precipitation; D006863:Hydrogen-Ion Concentration; D042845:Laccase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D020072:Polyporales / Q000201:enzymology*; D013451:Sulfonic Acids / Q000378:metabolism; D013696:Temperature
D052160:Benzothiazoles; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D042845:Laccase