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AuthorShrestha, Prabin
AuthorJoshi, Bishnu
AuthorJoshi, Jarina
AuthorMalla, Rajani
AuthorSreerama, Lakshmaiah
Available date2021-09-07T06:16:15Z
Publication Date2016
Publication NameBioMed Research International
ResourceScopus
ISSN23146133
URIhttp://dx.doi.org/10.1155/2016/3238909
URIhttp://hdl.handle.net/10576/22776
AbstractAt 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.
Languageen
PublisherHindawi Limited
Subject2 diethylaminoethanol
2,2' azinobis(3 ethylbenzothiazoline 6 sulfonic acid)
ammonium sulfate
copper sulfate
laccase
lignin
protein
sepharose
fungal protein
laccase
anion exchange chromatography
Article
controlled study
culture medium
delignification
enzyme activity
enzyme analysis
enzyme isolation
enzyme kinetics
enzyme purification
enzyme stability
enzyme substrate
enzyme synthesis
extracellular matrix
fractionation
fungal strain
fungus isolation
Ganoderma
Ganoderma japonicum
Ganoderma lucidum
habitat
Lentinula edodes
Nepal
nonhuman
pH
physical chemistry
protein expression
rice
straw
thermostability
chemistry
classification
ecosystem
enzyme activation
enzyme specificity
enzymology
Ganoderma lucidum
isolation and purification
microbiology
Oryza
species difference
Ecosystem
Enzyme Activation
Enzyme Stability
Fungal Proteins
Laccase
Nepal
Oryza
Reishi
Species Specificity
Substrate Specificity
TitleIsolation and Physicochemical Characterization of Laccase from Ganoderma lucidum-CDBT1 Isolated from Its Native Habitat in Nepal
TypeArticle
Volume Number2016
dc.accessType Open Access


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