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KMID : 0613820150250060673
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Journal of Life Science
2015 Volume.25 No. 6 p.673 ~ p.679
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Optimizing Culture Conditions to Maximize the Production of Laccase from Pholiota highlandensis
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Choi Hye-Ju
Moon Soo-Jung
Jeon Sung-Jong
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Abstract
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The culture conditions needed to maximize the production of laccase from Pholiota highlandensis mycelia were investigated. Among the tested media for laccase production, Coriolus versicolor medium (CVM; 2% dextrose, 0.4% peptone, 0.6% yeast extract, 0.046% KH2PO4, 0.1% K2HPO4, 0.05% MgSO4?7H2O) showed the highest activity for the enzyme. Then, to optimize culture conditions for laccase activity, the influences of various carbon, nitrogen, phosphorus, and inorganic salt sources in CVM were investigated. The optimum culture medium was 2% fructose, 0.4% peptone with 0.6% yeast extract, 0.05% NaH2PO4, and 0.05% MgSO4?7H2O as carbon, nitrogen, phosphorus, and inorganic salt sources, respectively. Several aromatic compounds in the medium enhanced laccase activity to varying degrees. Guaiacol induced maximum laccase production, yielding 114.1 U/ml laccase activity after cultivation for 11 days at 25°C. The optimum pH and temperature for laccase production were 8.0 and 35°C, respectively. Native polyacrylamide-gel electrophoresis (PAGE) followed by laccase-activity staining with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the presence of laccase under the optimum conditions studied. Zymogram analysis of the supernatant culture showed an enzymatic band with a molecular mass of about 90 kDa.
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KEYWORD
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Laccase, lignin, ligninolytic enzyme, Pholiota highlandensis, polyphenol oxidase
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