SATISH PRADHAN DNYANASADHANA COLLEGE ARTS, COMMERCE AND SCIENCE, THANE 2017-18 A Project Report on “ISOLATION AND SCREENING OF CELLULOLYTIC FUNGI FROM THANE CREEK ” Submitted by DR. MANDAKINI INGLE Department of Botany Satish Pradhan Dnyanasadhana College of Arts Commerce, Thane
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SATISH PRADHAN DNYANASADHANA COLLEGE
ARTS, COMMERCE AND SCIENCE, THANE
2017-18
A Project Report
on
“ISOLATION AND SCREENING OF CELLULOLYTIC
FUNGI FROM THANE CREEK ”
Submitted by
DR. MANDAKINI INGLE
Department of Botany
Satish Pradhan Dnyanasadhana College of Arts Commerce, Thane
INDEX
R. NO CONTENT PAGE NO.
01 Introduction 01-02
02 Review of Literature 03-04
03 Material and Method 05
04 Results 06-11
05 Discussion 12
06 Bibligraphy 13-14
CHAPTER 1
INTRODUCTION
Cellulolytic enzymes are synthesized by a number of micro-organisms. Fungi and bacteria are
the main natural agents of cellulose degradation (Lederberg, 1992). Fungi are well-known
agents of decomposition of organic matter, in general, and of cellulosic substrate in particular.
The initial step in cellulose degradation is enzymatic hydrolysis of polymers.
Cellulases are the third most significant commercial enzyme in the world market. Cellulases,
solely or in a mixture with other enzymes, involved in several industries including
biofuel,food, feed, beverages, agriculture etc (Kuhad et al.2011; Siti Lusi Sari et al. 2017).
Isolation and characterisationn of cellulolytic fungi provide a good starting point for the
discovery of beneficial enzymes (Rathnan et al.2012; Siti Lusi Sari et al. 2017). Most of the
cellulolytic fungi belong to Ascomycota and Basidiomycota phylums.
Mangroves are the coastal wetland forests generally found near the intertidal regions of
estuaries between creeks, lagoons, marshes etc. Mangroves provide a unique ecological site to
different microbes. Because of richness in carbon and other nutrients mangrove ecosystem
harbors diverse microbial communities which can adapt themselves in the extreme conditions
there. Microorganisms forms integral part of the mangrove ecosystem. Mangrove ecosystem
shows diversity of microbes such as bacteria, fungi, actinomycetes etc. The microorganisms of
mangrove are essential in the productivity, conservation, and rehabilitation of mangrove
ecosystems (Holguin et al., 2001).
The creek, known as “Thane Creek” separates the Island City of Mumbai in the west from the
mainland in the east and houses industrial areas at a distance of about 25 Km north-east of
Mumbai city. Thane Creek lies in the southern part of the Deccan belt of India between
latitude 18º 53’ to 19º 04’ N longitude 72º 48’ to 72º 53’E. It is a triangular mass of brackish
water which widens out and opens to the Arabian Sea in the South. The creek is narrow at the
Northern end, were it is fed partially by river Ulhas. Soil from thane creek is very rich in
micro-organisms like bacteria and fungi. Fungi can produce both intra-cellular as well as
extra-cellular enzymes. All fungi are hetrotropic, and rely on carbon compounds synthesized
by other living organisms.
Fungi as decomposers, play a most important role in our economy because fertility of soil
greatly depends on microbial activity.A wide variety of microbes including bacteria, fungi,
actinomycetes and protozoa are involved in the decomposition of cellulose, fungi have been
generally considered as main organisms’ responsible (Cowling, 1958). So the rate of
utilization of plant materials and its transformation into humus is largely dependent on the
active soil mycoflora. This is the reason why scientists are studying the fungi at molecular
level, trying to discover more cellulolytic fungi.
The aims and objectives of the present study include:
I) Isolation and identification of cellulose degrading fungi from soils of
Thane creek.
i) Preparation of large inoculum of each form to study cellulose
degradation capacity.
ii) Frequency distribution of cellulose decomposing fungi.
CHAPTER 2
REVIEW OF LITERATURE
Efficient hydrolysis of ligno-cellulose to simple sugars that can then be adequately
used as raw materials for production of ethanol and various industrial products by use
of chemicals is not cost effective (Sharada et al., 2013). Biological pretreatment of
lignocellulose via microorganisms is safe, environment friendly alternative (Okano et
al., 2005; Ravichandra et al., 2013). Many microorganisms are capable of degrading
and utilizing celluloses, hemicelluloses and other polysaccharides as carbon and
energy sources naturally in ecosystem. These organisms can produce a coordinate
battery of lingo-cellulolytic enzymes that are capable of degrading cellulose as well as
other plant cell-wall polymers. The lingo-cellulolytic enzymes comprises of a large
group of extracellular proteins including hydrolytic enzymes (cellulases.
hemicellulases, pectinases, amylases, mannases etc.) and lignolytic enzymes (Mtui,
2012).
Enzymatic hydrolysis of cellulose and other related oligo-saccharides is catalyzed by
cellulase. The cellulase enzyme system is a mixture of hydrolytic enzymes including
exoglucanases, endoglucanases and β-glucosidases acting in a synergistic manner
(Enari, 1983; Acharya et al., 2008).
Shobana and Maheshwari (2013) stated that cellulose producing fungi cultures using
agricultural wastes soil as substrate by culturing them on media plate and isolating
cellulolytic fungi for the study of maximum cellulase enzyme production activity by
the fungi.Fusarium oxysporum isolated from tomato plant parts produces maximum
cellulose at optimum parameters. (Ramananthan, etal., 2010).
Soma Mrudula and Rangasamy Murugammal ,have reported that the physical and
nutritional parameters of fermentation like pH, substrate, temperature, carbon and
nitrogen sources for cellulase production byAspergillus nigerduring submerged
fermentation and solid state fermentation play an important role. Kale and Zanwar
(2016) isolated Trichoderma and Aspergilluswere from different sources and screened
for their ability to degrade cellulose.Sumathi.et al.,(2016), investigated the efficacy of
fungal cultres isolated from plastic dumped soils as a substrate by using six different
culture mediums by serial dilution method. Cellulase producing fungi were isolated on
other specific culture media. Thorn et al ., (1996) screened 67 basidiomycetes from
40 of the 64 soil samples and 8 of 11 habitats and 17 of the 35 plots sampled.
Bhat and Shewade(2013), stated in their study by collecting mangrove soil samples
from five different stations of CBD Belapur, Navi Mumbai. Among the soil samples
screened, all gram positive isolates were obtained. 14 distinct isolates were
3. Bhat M. R. and Shewade, L. (2013). Isolation and Characteristics of micro-
organisms from mangrove soil of CBD Belapur creek, Navi Mumbai, MS India.4. C.J Alexopoulos et al., (1979). Book for Introductory mycology.5. Charitha Devi, M., and M.Sunil Kumar (2012). Production, Optimization and
Partial Purification of cellulase by Aspergillus niger fermented with paper and timber
sawmill industrial wastes, Journal of Microbiology and Biotechnology
Research,2(1):120-128.6. Gupta, N., Das, S. and Basak, U.C., (2007). Useful extracellular activity of
bacteria isolated from Bhitarkanika mangrove ecosystem of Orissa coast, Malaysian
Journal of Microbiology, 3(2):15-18.7. Kuhad, R.C., Singh, A. and Eriksson, K. E. (1997). Microorganisms enzymes
involved in the degradation of plant fiber cell walls. Adv. Biochem. Eng. Biotechnol.
57: 45-125.
8. Lowry, O.M., Rosebrough, N. J., Farr, A.L. and Randall, R.J. (1951). Protein
measurement with Folin-phenol reagent. Journal of Biological Chemistry. 193:265-
275.9. Ramanathan, G., Banupriya, S., and Abrirami, D. (2010). Production and
optimization of Cellulase from Fusarium oxysporum by Submerged
Fermentation.Journal of Scientific and Industrial Research. 69:454-459.10. Reese, E.T. and Mandels, M. (1984) Rolling with the time: production and
applications of Trichoderma reesei cellulase. Annual Report of Fermentation
Processes. 7:1–20.
11. Sahoo K. and Dhal, N.K. (2009), Potential microbial diversity in mangrove
ecosystem, A review Indian journal of Marine Sciences, 38(2): 249-256.
12. Sharada, R., Venkateswarlu, G., Venkateshwar, S. and Anand Rao, M. (2013).
Production of cellulase - A review. International Journal of Pharmaceutical, Chemical
and Biological Sciences, 3(4): 1070-1090.13. Singh, A., Singh, N. and Bishnoi, N.R. (2009). Production of Cellulases by
Aspergillus Heteromorphus from Wheat Straw under Submerged Fermentation.
International Journal of Civil and Environmental Engineering, 1:23-26.