SCREENING OF CELLULOLYTIC FUNGI FOR MANAGEMENT OF MUNICIPAL SOLID WASTE

Abstract

Currently, Rapid industrialization and Population explosion in India, the estimated quantity of the waste generation was11 billion tones of industrial wastes and 1.6 billion tones were municipal solid waste. About 90 billion tones solid wastes are expected to be generated annually by the year 2025. The main objective of this study is to maximize economic benefits and at the same time protection of environment by microbial activity for waste management. Cellulases are group of hydrolytic enzymes capable of hydrolyzing the most abundant organic polymer i.e. Cellulose to smaller sugar components including glucose subunits. Cellulases have enormous potential in industrials and used in food, beverages, textile, laundry, paper and pulp industries etc. This study was aimed to screen the cellulolytic ability of fungi from municipal solid waste. Out of 20 fungal culture isolates from environmental sources including 8 different zones, 16 fungi were found to passes cellulose degrading ability. Cellulolytic fungi belonging to Aspergillus funmigatus, Trichoderma sp.I and Chaetomium sp. Results obtained during this study clearly indicate that cellulase activity of Aspergillus fumigatus and Trichoderna sp.1 were found relatively towards the higher side and A. niger, A. flavus, A. nidulans, Alernaria sp., Penicillium sp. moderate range while Fusarium sp., Humicola sp. and Torula sp. showed low cellulase activity. These fungi were potential cellulose degrader and useful for the management of municipal solid waste.

Full text

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S.P.Gautam, P.S. Bundela, A.K. Pandey, M.K. Awasthi and S. Sarsaiya, 2010. Screening of Cellulolytic Fungi for
Management of Municipal Solid Waste.



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Journal of Applied Sciences in Environmental Sanitation, 5 (4): 391-395.









Research Paper

SCREENING OF CELLULOLYTIC FUNGI FOR MANAGEMENT OF MUNICIPAL
SOLID WASTE

S.P.GAUTAM1, P.S. BUNDELA2, A.K. PANDEY3, M.K. AWASTHI2 and S. SARSAIYA2*

1Central Pollution Control Board, New Delhi, India.
2Regional office M.P. Pollution Control Board Vijay Nagar Jabalpur (M.P.) India.
3M.P. Private Universities Regulatory Commission, Govt. of M.P. Bhoj University Campus, Kolar Road,
Bhopal (M.P.) India.
*Corresponding Author: Phone: +919826838221; Fax: 076124042780; E-mail: surendra_sarsaiya@yahoo.co.in

Received: 12th August 2010; Revised: 8th October 2010; Accepted: 29th October 2010

Abstract: Currently, Rapid industrialization and Population explosion in India, the
estimated quantity of the waste generation was11 billion tones of industrial wastes and
1.6 billion tones were municipal solid waste. About 90 billion tones solid wastes are
expected to be generated annually by the year 2025. The main objective of this study is
to maximize economic benefits and at the same time protection of environment by
microbial activity for waste management. Cellulases are group of hydrolytic enzymes
capable of hydrolyzing the most abundant organic polymer i.e. Cellulose to smaller
sugar components including glucose subunits. Cellulases have enormous potential in
industrials and used in food, beverages, textile, laundry, paper and pulp industries etc.
This study was aimed to screen the cellulolytic ability of fungi from municipal solid
waste. Out of 20 fungal culture isolates from environmental sources including 8 different
zones, 16 fungi were found to passes cellulose degrading ability. Cellulolytic fungi
belonging to Aspergillus funmigatus, Trichoderma sp.I and Chaetomium sp. Results
obtained during this study clearly indicate that cellulase activity of Aspergillus fumigatus
and Trichoderna sp.1 were found relatively towards the higher side and A. niger, A.
flavus, A. nidulans, Alernaria sp., Penicillium sp. moderate range while Fusarium sp.,
Humicola sp. and Torula sp. showed low cellulase activity. These fungi were potential
cellulose degrader and useful for the management of municipal solid waste.

Keywords: Cellulosic material, Thermogenesis, Municipal solid waste (MSW), Cellulases.





ISSN 0126-2807


Volume 5, Number 4: 391-395, October-December, 2010
© T2010 Department of Environmental Engineering
Sepuluh Nopember Institute of Technology, Surabaya
& Indonesian Society of Sanitary and Environmental Engineers, Jakarta
O p e n A c c e s s h t t p : / / w w w . t r i s a n i t a . o r g



This work is licensed under the Creative Commons Attribution 3.0 Unported License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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S.P.Gautam, P.S. Bundela, A.K. Pandey, M.K. Awasthi and S. Sarsaiya, 2010. Screening of Cellulolytic Fungi for
Management of Municipal Solid Waste.

INTRODUCTION

The large volumes of cellulosic waste generated annually from forestry, commercial,
agricultural and industrial activities are difficult to degrade and cause imbalances in the
ecosystem. These residues encompass material such as agricultural residue, municipal solid
waste (MSW), cellulosic material (35-45%) found in MSW. Cellulosic residues are renewable
resources of which over a billion tons are generated annually in world [1]. The annual production
of cellulose is estimated to be 4.0X107 tons [2]. Treatment of MSW with either cellulolytic
enzymes or microorganism could lead to more efficient degradation of this waste material,
promote cycling of nutrients in the environment and reduce the impact of waste accumulation on
terrestrial and aquatic ecosystem.
Cellulases are consortia of complex hydrolytic enzymes capable of hydrolyzing these
materials to smaller sugar components like glucose units. This enzyme system which shows
excellent degradative action towards crude natural cellulosic material is routinely found in most of
the fungi and essentially comprises three main components namely exoglucanase,
endoglucanase and β-glucosidase [3]. A large number of fungi have been isolated by various
investigators from self heating material and other sources play an important role in decomposing
of municipal solid waste at elevated temperature that usually resulted from microbial
Thermogenesis [4]. Fungi capable of producing extracellular enzymes responsible for
degradation of cellulose are known, some of them being highly cellulolytic, which include species
of Aspergillus sp., Trichoderma sp., Sclerotium sp. and are also being considered for
commercially exploitation [5-10].
The present study was initiated to isolate and screen the indigenous fungal strains and to
explore their hydrolytic potential for their possible future application.

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Journal of Applied Sciences in Environmental Sanitation, 5 (4): 391-395.

MATERIALS AND METHODS

Collection of Samples
Total 162 Samples of MSW were collected from 8 different localities of Jabalpur: Zone 1,
Zone 2, Zone 3, Zone 4, Zone 5, Zone 6, Zone 6, Zone 7 and Zone 8. The samples were taken
by means of sterilized spatulas and collected in sterile sealed polythene bags. The samples were
then brought to the laboratory for microbiological study.

Isolation of Fungi
All the samples were processed within 3 hrs of collection. For isolation, dilution plate method
was used [11]. One (1) gm fresh weigh sample was placed in 10 ml of sterile water and shaken
for proper mixing at room temperature. Then, 100 µl portions of the suspensions were inoculated
onto plates containing potato dextrose agar (25% Potato, 1.5% Agar (w/v). Chloramphenicol was
added to the molten medium after autoclave, and the plates were incubated at 28±20C for 4-8
days to identify the fungi. All the isolated from Jabalpur MSW were identified according to [12]
and maintained on PDA slants at low temperature (4±1oC) for further study.
Screening for Cellulase Enzyme
From the various isolates, screening for cellulolytic fungi was made using PDA media
supplemented with 5% carboxymethyl cellulose (CMC). Cellulolytic fungi create a clearing zone
around the colony on the agar [13].

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S.P.Gautam, P.S. Bundela, A.K. Pandey, M.K. Awasthi and S. Sarsaiya, 2010. Screening of Cellulolytic Fungi for
Management of Municipal Solid Waste.

RESULTS AND DISCUSSION

The numbers of fungi isolated and details of the samples collection from the various locations
are shown in Table 1 clearly indicate that the highest number of fungi were isolated from zone 4
(21), zone 1 (16) and zone 6 (14). Among the fungi isolates, only five were identified to the
species level presented in the table 2. It was fund that fungi isolated, respectively were obtained
from 8 different zone of the Jabalpur city were potential cellulose degrader. Twenty different
isolates were screened for their ability to produce extracellular cellulases. The cellulase
production ability of fungi assessed by estimating zone around the colony formed due to ability of
fungal isolates to hydrolyse cellulose.

Table 1: Details of sample collection and fungal isolates
Zones
of
Jabalpur
Zone 1 54 20 12
Zone 2 15 2 11
Zone 3 10 1 4
Zone 4 25 4 1
Zone 5 10 4 4
Zone 6 18 11 2
Zone 7 15 2 6
Zone 8 15 1 5
Total 162 45 45

Results presented in Table 2 indicate that only sixteen isolates were able to synthesize
extracellular cellulase. Highest cellulolytic activity was detected in only seven isolates viz.
Trichoderma sp.I (4.7mm), Chaetomium sp. (2.7mm), Aspergillus fumigatus (2.3mm), A. flavus
(1.7mm), A. nidulans (1.5mm), Trichoderma sp.II (1.5mm) and A. niger (1.4mm). The enzymatic
activity was considerably low in other fungi such as Aspergillus sp. (0.2mm), Curvularia sp. I
(0.8mm), Curvularia sp. II (1.3mm), Fusarium sp. (0.6mm), Fusarium oxysporum (0.5mm),
Humicola sp. 0.2mm), Penicillium sp. 0.9mm) and Torula sp. 0.5mm) while Alternaria sp.,
Cladosporium sp., Exserohilum sp. and Helminthosporium sp. were not produce cellulase
enzymes. Aspergillus sp. was isolated from all the substrates and its occurrence was also
predominantly high among other fungal strains followed by Fusarium sp. Curvularia sp., and
Trichoderma sp.
Only few strains of Trichoderma sp., which is the most extensively studied cellulase producer
were isolated in this study. This may be due to a possibility that the growth of Trichoderma sp.
was suppressed by rapidly growing fungi like Aspergillus sp. Majority of Aspergillus sp.,
Trichoderma sp.I Isolates were found to posses cellulolytic activity. The present study indicates
that number of fungal forms inhabiting various cellulolytic wastes produce extracellular cellulases.
The screened fungal strains will be useful for further studies by enzyme producers.
Fungi are known agents of decomposition of organic matter in general and of cellulosic substrate
in particular as reported by [14]. Trichoderma sp. is well known among the cellulolytic fungi for
their potential to degrade organic municipal solid waste. A few studies have been conducted
earlier with Trichoderma sp. to investigate their cellulolytic ability [15]. Screening of different
genera species and even isolated is first step for finding acceptable enzyme producer isolated.

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Journal of Applied Sciences in Environmental Sanitation, 5 (4): 391-395.

Number of
samples
Types of material and number of sample
Plant residue Compost
Fungal
isolates
Soil Mixed MSW
14
1
3
5
1
2
5
8
39
8
1
2
15
1
3
2
1
33
16
2
5
21
9
14
11
9
87

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S.P.Gautam, P.S. Bundela, A.K. Pandey, M.K. Awasthi and S. Sarsaiya, 2010. Screening of Cellulolytic Fungi for
Management of Municipal Solid Waste.

[16] in surveys on some genera like Trichoderma sp., Fusarium sp., Aspergillus sp.,
Phanerocheate sp., Chrysosporium sp. and Sclerotium sp. showed some differences in their
cellulase activity. Members of the fungal genus Trichoderma sp. and Aspergillus sp. have been
extensively studied, particularly due to their ability to screte cellulose degrading enzyme the most
common and most potent cellulase producers are Trichoderma ressei, T. koningii, Fusarium sp.
Table 2 clearly depicts that Trichoderma sp.I, Aspergillus fumigatus showed excellent cellulose
producing results in the screening. Morederate cellulose producers were also recorded among
other fungi. The present finding indicates that the cellulose system of these fungal forms contain
enzymes complexes for the effective hydrolysis of cellulose [17,18].

Table 2: Screening and measurement of zone of clearance by fungal isolates
S. No. Fungi
1. Aspergillus niger
2. Aspergillus flavus
3. Aspergillus fumigatus
4. Aspergillus nidulans
5. Aspergillus sp.
6. Alternaria sp.I
7. Alternaria sp.II
8. Chaetomium sp.
9. Curvularia sp.I
10. Curvularia sp.II
11. Cladosporium sp.
12. Exserohilum sp.
13. Fusarium sp.
14. Fusarium oxysporum
15. Helminthosporium sp.
16. Humicola sp.
17. Penicillium sp.
18. Torula sp.
19. Trichoderma sp.I
20. Trichoderma sp.II

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Journal of Applied Sciences in Environmental Sanitation, 5 (4): 391-395.


CONCLUSION

It may be concluded from the foregoing that the successful colonization of fungi as decaying
MSW in past may be correlated with their cellulolytic ability. The result of using different cellulosic
materials as substrate for the isolation of diverse group of fungi showed that the large number of
cellulolytic fungi present Zone 1, Zone 4 and Zone 6 isolated a number of fungi from self heating
materials.
Acknowledgements: The authors are thankful to M.P. Pollution Control Board Bhopal and Head,
Department of Biological Sciences, R.D. University, Jabalpur, for laboratory facilities. Also thanks to
Municipal Corporation of Jabalpur for his support. Ministry of Environment and Forest New Delhi is also
thankfully acknowledged for financial support.



Zone of Clearance in mm
1.4
1.7
2.3
1.5
0.2
1.1
0.0
2.7
0.8
1.3
0.0
0.0
0.6
0.5
0.0
0.2
0.9
0.5
4.7
1.5

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S.P.Gautam, P.S. Bundela, A.K. Pandey, M.K. Awasthi and S. Sarsaiya, 2010. Screening of Cellulolytic Fungi for
Management of Municipal Solid Waste.

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