Isolation and Physicochemical Characterization of Laccase from Ganoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal.

At present, few organisms are known to and capable of naturally producing laccases and white rot fungi are one such group. In the present study, three fungal species, namely, Ganoderma lucidum-CDBT1, Ganoderma japonicum, and Lentinula edodes, isolated from their native habitat in Nepal were screened...

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Publicado en:BioMed Research International Vol. 2016; pp. 1 - 11
Autores principales: Shrestha, Prabin, Joshi, Bishnu, Joshi, Jarina, Malla, Rajani, Sreerama, Lakshmaiah
Formato: pictorial research tables/charts Journal Article
Publicado: Wiley-Blackwell 10/16/2016
Acceso en línea:Ver este registro en EBSCOhost
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      dt: 10/16/2016
      vid: 2016
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      pub: Wiley-Blackwell
      place: Malden, Massachusetts
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        118823536
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        10.1155/2016/3238909
        118823536
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        atl: Isolation and Physicochemical Characterization of Laccase from Ganoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal.
      aug:
        au:
          Shrestha, Prabin
          Joshi, Bishnu
          Joshi, Jarina
          Malla, Rajani
          Sreerama, Lakshmaiah
        affil: Central Department of Biotechnology, Tribhuvan University, Kirtipur, Nepal
      sug:
        subj:
          Oxidoreductases Analysis
          Fungi Analysis
          Nepal
          Descriptive Statistics
          Chromatography Methods
          Chemistry, Analytical
          Hydrogen-Ion Concentration
          Kinetics
          Fungi Classification
          Cellulose
          Biological Assay
          Electrophoresis Methods
          Data Analysis Software
          Culture Media
          Temperature
      ab: At present, few organisms are known to and capable of naturally producing laccases and white rot fungi are one such group. In the present study, three fungal species, namely, Ganoderma lucidum-CDBT1, Ganoderma japonicum, and Lentinula edodes, isolated from their native habitat in Nepal were screened for laccase production, and G. lucidum-CDBT1 was found to express highest levels of enzyme (day 10 culture media showed 0.92 IU/mg total protein or 92 IU/mL laccase activity with ABTS as substrate). Lignin extracted from rice straw was used in Olga medium for laccase production and isolation from G. lucidum-CDBT1. Presence of lignin (5 g/L) and copper sulfate (30 μM) in the media increased the extracellular laccase content by 111% and 114%, respectively. The laccase enzyme produced by G. lucidum-CDBT1 was fractionated by ammonium sulfate and purified by DEAE Sepharose anion exchange chromatography. The purified enzyme was found to have a molecular mass of 43 kDa and exhibits optimal activity at pH 5.0 and 30°C. The isolated laccase was thermally stable for up to 70°C for 1 h and exhibited broad pH stability. The kinetic constants, Km, Vmax, and Kcat, determined using 2,2′-azinobis-(-3-ethylbenzothiazoline-6-sulfonic acid) as substrate were found to be 110 μM, 36 μmol/min/mg, and 246 min−1, respectively. The isolated thermostable laccase will be used in future experiments for delignification process.
      pubtype: Academic Journal
      doctype:
        pictorial
        research
        tables/charts
        Journal Article
      ougenre: Article
    language: English
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