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Life Science Research and Sustainable Development ISBN: 978-98-84663-33-9

from compost soil and characterize them. 11 fungal isolates were isolated by serial dilution and
they were cultivated on potato dextrose agar plate with indicator compound ABTS to screen for
the laccase production ability. Out of 11 isolates, one was presumed to be potent, another showed
medium potency, and three showed weak laccase producing ability.
Daphne and Gnanadoss (2013) used two substrates viz. Guaiacol and ABTS in media to
isolate laccase producing fungi. Twenty six fungi were isolated and screened for their ability to
produce laccase on solid medium containing guaiacol and ABTS. Six isolates showed positive
laccase activity out of that two fungi were quantitatively screened and identified as Psathyrella
condolleana LCJ 178 and Myrothecium gramineum LCJ 177. Kaur and Nigam (2014) in search of
active producer of laccase enzyme used different culture media out of tested media, malt extract
medium supplemented with substrate ABTS showed maximum laccase production out of other
tested media and substrates or indicator compounds. Abeer et al., (2015) carried out qualitative
and quantitative assay of marine fungal isolate Alternaria tenuissima and reported that it showed
highest zone diameter and colony diameter in agar plate screening test with guaiacol as an
indicator compound.
Vantamuri and Kaliwal (2015) isolated 150 fungal strain and screen for their ability to
produce laccase enzyme using ABTS, tannic acid, guaiacol and syringaldazine as an indicator
compounds and showed that among 150 fungi 8 fungal strain shown ability for laccase
production. Fatima et al., (2015) screened soil fungi for laccase production by using guaiacol as a
substrate and found that Verticillum sp. and Helminthosporium sp. showed large reddish-brown
zone in media due to oxidative polymerization of guaiacol. Sidhu et al., (2017) isolated twenty
five lignolytic fungi from decaying wood samples and screened for the laccase production using
guaiacol and ABTS as indicator compounds. The positive isolates were confirmed for the
presence of laccase using indicators and quantitative enzyme assay in presence of catalase using
ABTS as substrate.
6. Optimization of culture condition for laccase production:
6.1. Optimization of proper culture media for laccase production:
Researcher used different medium for laccase production such as; Potato Dextrose Broth,
Malt Extract Broth, Glucose Peptone Broth, Glucose Nitrate Broth, Yeast Extract Peptone Dextrose
Broth and Czapek Dox Broth.
Manimozhi and Kaviyarasan (2012) concluded that malt dextrose broth and potato
dextrose broth was optimal medium for laccase activity and biomass production of Agaricus
heterocystis. Amutha and Abhijit (2015) studied different media for growth and laccase production
from Trametes versicolor and observed that yeast glucose broth is suitable for the growth of
Trametes versicolor at pH 5.5 and temperature 28 C and Modified Kirk and Ferrell media showed
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maximum enzyme production in presence of pH 5.5 and temperature at 28 C. Prasher and
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Chauhan (2015) carried out an experiment, effect of twelve different basal media for the growth
and ligninolytic enzymes activity of Dictyoarthrinium synnematicum and reported that the
Glucose-peptone medium was found to be an optimum medium for the growth and laccase
production of Dictyoarthrinium synnematicum. Sidhu et al., (2017) studied laccase producing
capacity of fungus estimated in seven different media and reported that Czapek dox medium
showed highest enzymatic activity.
6.2. Effect of different nutrient sources on laccase production:
Some researchers observed that the elevated laccase activity was achieved by using low
carbon to nitrogen ratio while others observed that it was achieved at high carbon to nitrogen
ratio.

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