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Research article

J. Microbiol. Biotechnol. 2011; 21(12): 1312-1321

Published online December 28, 2011 https://doi.org/10.4014/jmb.1106.06036

Copyright © The Korean Society for Microbiology and Biotechnology.

Chitinolytic and Chitosanolytic Activities from Crude Cellulase Extract Produced by A. niger Grown on Apple Pomace Through Koji Fermentation

Gurpreet Singh Dhillon 1, Satinder Kaur Brar 1*, Surinder Kaur 1, 2, Jose R. Valero 1 and Mausam Verma 3

1INRS-ETE, Université du Québec, 490, Rue de la Couronne, Québec, G1K 9A9, Canada, 2Department of Mycology & Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University (BHU), Varanasi-221005, India, 3Institute for Research and Development in Agri-Environment (IRDA), 2700 rue Einstein, Quebec (Quebec), G1P 3W8, Canada

Received: June 16, 2011; Accepted: August 5, 2011

Abstract

Enzyme extracts of cellulase [filter paper cellulase (FPase)
and carboxymethyl cellulase (CMCase)], chitinase, and
chitosanase produced by Aspergillus niger NRRL-567
were evaluated. The interactive effects of initial moisture
and different inducers for FP cellulase and CMCase
production were optimized using response surface
methodology. Higher enzyme activities [FPase 79.24±
4.22 IU/gram fermented substrate (gfs) and CMCase
124.04±7.78 IU/gfs] were achieved after 48 h fermentation
in solid-state medium containing apple pomace supplemented
with rice husk [1% (w/w)] under optimized conditions [pH
4.5, moisture 55% (v/w), and inducers veratryl alcohol
(2 mM/kg), copper sulfate (1.5 mM/kg), and lactose 2%
(w/w)] (p<0.05). Koji fermentation in trays was carried
out and higher enzyme activities (FPase 96.67±4.18 IU/gfs
and CMCase 146.50±11.92 IU/gfs) were achieved. The
nonspecific chitinase and chitosanase activities of cellulase
enzyme extract were analyzed using chitin and chitosan
substrates with different physicochemical characteristics,
such as degree of deacetylation, molecular weight, and
viscosity. Higher chitinase and chitosanase activities
of 70.28±3.34 IU/gfs and 60.18±3.82 to 64.20±4.12 IU/gfs,
respectively, were achieved. Moreover, the enzyme was
stable and retained 92-94% activity even after one month.
Cellulase enzyme extract obtained from A. niger with
chitinolytic and chitosanolytic activities could be potentially
used for making low-molecular-weight chitin and chitosan
oligomers, having promising applications in biomedicine,
pharmaceuticals, food, and agricultural industries, and in
biocontrol formulations.

Keywords: cellulase, chitooligomers, chitinase, chitosanase, solid-state tray fermentation, response surface methodology