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Fomitopsis pinicola 균사체로부터 Laccase의 최적생산조건

Optimal Conditions for the Laccase Production from Fomitopsis pinicola Mycelia

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초록

The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose,0.2% peptone, 0.2% yeast extract, 0.05% KH2PO4, and 0.05% MgSO4 · 7H2O) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% NaH2PO4 and 0.05% CaCl2 were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at 25℃. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were 80℃ and pH 3.0.

Abstract

The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose,0.2% peptone, 0.2% yeast extract, 0.05% KH2PO4, and 0.05% MgSO4 · 7H2O) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% NaH2PO4 and 0.05% CaCl2 were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at 25℃. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were 80℃ and pH 3.0.

참고문헌 (36)

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