CHAPTER ONE

INTRODUCTION
1.1 Background of the Study
Pectinase is a group of enzymes that catalyze a degradation of pectin through depolymeration
(hydrolases and lyases) and de-esterification (esterases) reactions (Pedrolli et al., 2009). Based
on their mode of action, the pectinase is classified into three types: pectin esterase, hydrolases
and lyases. Pectin esterase catalyze the de-esterification of the methoxyl group of pectins,
forming pectic acid. Hydrolases (polygalacturonases and polymethylgalacturonases) catalyze the
hydrolytic cleavage of α-1, 4glycosidic linkage in pectic acid and pectin. Lyases
(polygalacturonate lyase and polymethylgalacturonate lyase) catalyze the cleavage of α -1, 4-
glycosidic linkage in pectic acid and pectin, forming unsaturated galacturonates and methyl
galacturonates (Garg et al., 2016).
Pectinases share about 25% in global sales of food enzymes (Tapre and Jain, 2014). Pectinases
are widely used in food industries, such as fruit processing industry for extraction and
clarification of fruit juice, preparation of fruit cordials (Merín et al., 2015), enhance process
efficiency of wine making and wine quality (Merín et al., 2015), oil extraction and fermentation
of coffee, cocoa and tea (Tapre and Jain, 2014). Since pectinases are widely used enzyme for
different industrial application, it is necessary to improve pectinase production, including
selection for potential source of pectinase. Many experiments have attempted to increase the
performance of enzyme production (Molina et al., 2011). The new microbes with high
extracellular pectinase activity, stability over wide range of temperature and pH for a longer
period of time, with their cost-effective production have been the focus of recent research (Garg
et al., 2016). Mango peel Mango is an important tropical fruit, cultivated in many tropical
regions and distributed worldwide. India is largest producer of mango amongst 90 countries in
the world, with its production of 18, 643, 000 metric tons on 2, 209, 000 hectare area which is
35.8% of total fruit area (DACFW 2015; Reddy and Saritha 2015). Mango peel, one of the major
by-products from the mango pulp industry, constitutes about 20–25% of the mango fruit
processing waste and is discarded during the process. It is a rich source of pectin, which
comprises 12.2–21.2% pectin with degrees of esterification from 56.3 to 65.6% (Cheok et
al. 2016). Mango peel has been used in SSF for pectinase production by A. foetidus (Kumar et
al. 2012). The substrate mango peel was supplemented with salt solution consisting of (g/l)

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(NH4)2SO4, 25; MgSO4, 0.6; FeSO4, 0.4; urea, 3; peptone, 5 and KH2PO4, 6 at pH 7.0 for use in
SSF. Reddy and Saritha (2016) also used mango fruit processing waste (0.5%) for production of
pectinase by Enterobacter sp. PSTB-1in SmF.
Pectinolytic enzymes are a group of related enzymes that hydrolyse pectic substances. Pectin is a
complex polysaccharide present in the middle lamella of plant cell walls. It is composed of
multiple units of D-Galacturonic acid linked by α (1, 4) glycosidic linkage. Pectinolytic enzymes
have been reported in higher plants (Nighojkar et al. 1994; Jolie et al. 2010) and microorganisms
including bacteria and fungi (Uzuner and Cekmecelioglu 2015; Patidar et al. 2016; Rebello et
al. 2017). Pectin is completely digested by three major enzymes: pectin methylesterase
(pectinesterase; EC: 3.1.1.11), pectinase (polygalacturonase; EC: 3.1.1.15) and pectin lyase (EC:
4.2.2.10) to release galacturonic acids and its oligomers (Combo et al. 2012). In nature,
microorganisms have been endowed with vast potential. They produce a range of enzymes,
which have been exploited commercially over the years. It has been reported that microbial
enzymes account for 25% of total global enzyme sales (Jayani et al. 2005). The pectic substances
can be converted by means of microorganisms or their enzymes into constituent
monosaccharides or specific oligosaccharides without the production of undesirable by-products
(Zykwinska et al. 2008; Martínez et al. 2009).
Pectinases are known for their tremendous potential in various industries. Pectin methylesterase
and endo-polygalacturonase have important role in softening of fruits, extraction and
clarification of juices, preparing gel, food manufacturing, retting of textile fibers, extraction of
olive oil, protoplast isolation, etc. (Kashyap et al. 2001; Jayani et al. 2005; Kohli and
Gupta 2015). Galacturonic acid, produced by action of pectinolytic enzymes, has various
applications in industries mainly in pharmaceutical industries. It is used for the production of
vitamin C as acidic agent in food industries and as washing powder agent in chemical industries
(Molnar et al. 2009; Burana-Osot et al. 2010). Almost all the commercial preparations of
pectinases are produced from fungal sources (Kertesz 1951). Filamentous fungi
especially Aspergillus niger is the major producer of acidic pectinase used mainly in fruit juice
and wine industries (Kashyap et al. 2001).
The utilization of SSF processes is interesting for pectinase production by fungi and has the
advantages of low water requirement, high productivity, lesser chance of contamination, cost
effectiveness and simpler fermentation technology (Pandey et al. 2000; Viniegra-González et

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al. 2003). Research on the selection of suitable solid substrates for SSF has mainly been centered
on agricultural and industrial residues due to their potential advantages for filamentous fungi,
which are capable of penetrating into the hardest of these solid substrates. In addition to the
utilization of this agro-industrial waste, it provides alternative substrates and helps in solving the
pollution problems (Pandey 2003).
Due to the wide applications of polygalacturonase and pectin methylesterase, there is a need to
highlight recent developments on several aspects related to their production in SSF. Microbial
sources, production, characterization and application of pectinases have been reviewed (Kashyap
et al. 2001; Jayani et al. 2005; Favela-Torres et al. 2006; Sharma et al. 2013). However, the most
common sources of pectinolytic microorganisms, recent development in SSF process for
polygalacturonase and pectin methylesterase production, and their assay methods have not been
considered until now.
1.2 Statement of the Problem
Mango (Mangifera indica) contain substantial amount of pectin having a high gelling grade
(Bhardwaj and Garg, 2010). In the processing of mango products, mango peels is a major by-
product which ends up as a waste product. However, mango peels can be used as a valuable,
economic and abundant media source for the commercial production of natural enzymes such as
pectinases. Pectin acts as an inducer for the production of pectinolytic enzymes by microbial
systems. Mango peel is one of the major by-products from the mango pulp processing industries.
During the processing of mango fruit, peel and stone are generated as waste (40–50% of total
fruit mass). Waste generated from mango processing constitutes 20–25% peel, which was found
to be a good source for the extraction of pectin of good quality, with a high degree of
esterification and phenolic compounds (Berardini et al., 2005). Pectin acts as the inducer for the
production of pectinase enzymes by microbial systems, and pectin-rich mango Peels are
considered to be a good source for pectinase production and ideal substrate for the
decomposition of mango peels by microorganisms (Kumar et al., 2012). The current commercial
sources of pectinase comprise a wide variety of bacteria, yeast and filamentous fungi (Yazid et
al., 2011). Amande and Adebayo-Tayo (2012) noted that among the filamentous fungi
Aspergillus spp. are the strains of choice for polygalacturonase production.
1.3 Justification

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Pectinases are widely used in food industries for juice extraction and clarification, coffee and tea
fermentation, oil extraction, improvement of chromacity and stability of red wines. This research
project will help to determine pectinolytic filamentous fungi from decomposed mango peels.
These produce a huge amount of waste throughout the year. These waste materials can be
constructively used to produce important products including pectin and pectinases as a
replacement for dumping them in the environment where they might at times generate pollution.
This is a promising way of converting wastes to wealth.
1.4 Aim and Objectives of the Study
1.4.1 Aims
The aim of this study is to isolate and characterize of pectinolytic filamentous fungi from
decomposed mango peels..
1.4.2 Objectives:
i. To isolate pectinases from decomposed mango peels samples
ii. To screen for the ability of yeast isolates to ferment pectinases.
iii. To study the physiological characteristics of selected isolates from decomposed mango
peels.