Review of Related Literature

Mango ( Mangifera indica L.) is rich in pectin content (Tandon and Garg, 1999), and its
peel offers a suitable growth medium for pectinolytic microorganisms. In recent study, the
common inhabitant in the mango surface was the genus Aspergillus, and they can be isolated
almost of all the mango cultivars (Pelczar et.al, 2008).
Naidu and Panda (1998) states that Aspergillus niger has the highest enzyme activity and
this is the most commonly used fungus for the industrial production of pectinolytic enzymes.
The mango peel contains a pectin content of 18.2 (w/w) and it can be used as a substrate
for pectinase enzyme. With Solid state fermentation, Aspergillus foetidus shows extreme
production of polygalacturonase and pectin lyase had optimum activities at pH 5 and 5.5,
temperatures 35C and 30C respectively. The enzymes are utilized for mango juice processing
for clarification and it shows maximum clarification of about (92.50.26%) at a temperature of
about 40C and 150 minutes of incubation on mango pulp (Yannam et.al,2012). The fungal
isolate Aspergillus sclerotioniger produced polygalacturonase in the highest value on the 6 day
of fermentation ranged from 0.448 4.7745 U/ml using mango peel as substrate on submerged
fermentation (Tiukaa et.al,2012).
Juices, nectars concentrates, jams, jelly powders and flakes are some products in
processing and utilization of mango fruit, whereas waste are generated in the form of peels and
kernels (Udenwobele et.al,2014).
Pectinases are a group of enzymes that catalyze the breakdown of pectins. They can be
divided in polygalacturonase (PG), pectin lyase (PL) and pectate lyase (PE). The synergistic
action of methyl esterase, endo-polygalacturonase, exopolygalacturonase endopectate lyase
completes the degration of pectin (Kuhad et.al,2004)

Commercially,microbial pectinase is one of the promising enzymes recently due to their

economic value (Darah et.al,2013).
Pectinases are posh cluster of enzymes which degrade different pectic substances present
in plant tissues. These enzymes have useful applications in paper, fruit and textile industries.
Almost 75% of the estimated sale value among industrial enzymes in 1995 has been contributed
by pectinases (Kashyap, Vohra, & Tewari, 2001).
According to Zhong & Cen (2005), these enzymes also have biological importance in
identification of plant diseases and protoplast fusion technology. Microorganisms have more
number of advantages and can be used for enzymes production at a higher level. Pectinases have
great biotechnological potential and can be used in many industrial processes. Hence,
applications of pectinases in various fields are widening, it is necessary to understand the
properties and nature of these enzymes for efficient and effective usage(Sathyanarayana
et.al,2003). Pectinase enzymes are yield from a vast variety of microbial sources such as fungi,
bacteria, yeast.
Pectinases are employed in many industrial processes includes fruit juice extraction,
textile processing, degumming of bast fibres, paper industry , treatment of industrial waste water,
coffee and tea fermentation etc., Pectinolytic enzmes were used in clarification of fruit juice
(Blanco et al., 1999), treatment to banana, apple, grapes, increased of juice volume of apple fruit
pulp (Kaur et al., 2004), promotes excessive softening during the process of fermentation and
storage (Baker & Wicker, 1996), accelerates the tea fermentation process and also destroys foam
foaming property (Carr, 1985) and used to eliminate mucilaginous coat from coffee beans
(Ranveer et.al,2005) .

Submerged (SMF) and Solid-State fermentation (SSF) are widely used for pectinase
production by different microorganisms. Some factors that influences the microbial pectinase
produced by SMF are the type and concentration of the carbon source , the culture pH, the
oxygen concentration (Rashmi et.al,2008)
In bioconversion of agro-industrial wastes such as orange peels, hydrolysis of polymers is
essential for their breakdown to monomers which can further be fermented to ethanol (as an
alternative to energy source) and other products. Two major polymers needed to be hydrolysed in
orange peels are pectins and celluloses. The enzymatic hydrolysis of pectinolytic and
cellulolytic substances requires synergistic actions of both pectinolytic and cellulolytic enzymes
(Grohmann and Baldwin, 1992).
Pectinases are widely used for biotechnological applications in food industry (fruit juice
extraction, coffee and tea fermentation, oil extraction, improvement of chromaticity and stability
of red wines), textile, paper and pulp industries (Cao et al., 1995). Pectinase production
has been reported from bacteria including Actinomycetes (Bruhlmann et al., 1994; Elegado et
al., 1999; Beg et al., 2000), Yeasts (Huang and Mahoney, 1999) and Fungi (Hawksworth et al.,
1983; Gummadi and Panda, 2003). However, for the industrial production of pectinases,
Aspergillus niger strains are used exclusively (Marcia et al., 1999). The enzyme preparations
used in the food industry are of fungal origin because fungi are potent producers of pectic
enzymes and the optimal pH of the fungal enzymes are very close to the pH of many fruit
juices, which ranges from pH 3-5.5. Such preparations are not suited for the production of
vegetable purees or other preparations in which pH values are close to neutral. Therefore, the
commercial pectinase production is still dominated mainly by Aspergillus niger strains
Pectinases are a heterogeneous group of enzymes that degrade pectin. These are

widely used in the food industry for the production and clarification of fruit juices, to
improve the cloud stability of fruit and vegetable juices and nectars, for depectinization in order
to produce high density fruit juice concentrates, and for haze removal from wines Pectic enzyme
preparations are also used for the production of low methoxypectin for
diabetic foods, in the degumming of natural fibers in the textile industry, and in making
commercial softwoods, such as Sitka and Norway spruce, more permeable to preservatives .
Purified pectinases have also been developed specifically for use in plant protoplast culture
studies. When used with cellulase, purified pectinases have been found to be very useful for
generating good yields of viable protoplast in several plant systems, e.g. corn, soybean, red beet,
sunflower, tomato, citrus etc. Commercial enzymes are generally obtained from fungal sources
since the pH optima of these enzymes are in the range found naturally in materials to be
processed and the enzymes are secreted into the culture media, making the downstream
processing easier. Keeping in view the importance of enzyme
pectinases in the food processing industry and the problems associated with the
disposal of food processing industry waste, the present study was undertaken with the
objectives of utilizing citrus peel for the production of pectinase(Bhardwaj and garg,2011).
Pectin, a polymer of galacturonicacid residues connected by a-1, 4 glycosidic linkages,
being the main component of middle lamella of plant cell wall. Pectinase enzyme hydrolyse
pectic substances into sugar which can be used for food and value added products. Pectinase are
industrially important enzymes and have potential applications in fruit, paper, textile, coffee and
tea fermentation industries. Pectinases are either intracellular or extra cellular. Although a large
number of micro-organisms can degrade pectin. Keeping in view, the demand of new enzymes

some micro-organism capable of pectinase production are being isolated and studied (Bhardwaj
and Garg,2011).
Pectinases or Pectinolytic enzymes are the one which have broadest applications
in the food processing, alcoholic beverages and textiles industries. These enzymes are chiefly
produced from the plants and microorganisms. The fruit peels are regarded as waste by most of
the industries. And the disposal of them becomes the serious problem, as it leads to the
environmental pollution. On the other hand, it is of low-cost and it contains pectin, a natural
substrate that contains selective chemical compound which is suitable for the production of
pectinase enzyme.
This review mainly concerned about the selection of substrate as peels and the production of
pectinolytic enzymes using different fruit peels, comparison of fermentation method that is
suitable for enzyme production using peels as substrates, different enzyme assay methods,
computer

software

controller

for

fermentation

used

and

also

applications

of

pectinase(Kumar,2014).
The Solid State Fermentation (SSF) is a method that primarily involves the growth of
microorganisms on a wet solid supports in the absence (or close to absence) of free water. New
interest in this technique derives from merely possible fact that it is considered to be an actual
approach for the processes that includes the bioremediation as well as removal of toxic substance
from agricultural wastes, biopulping and biotranformation of crops etc., [24,25,26]. The SSF
uses a different variety of natural solid supports that supply the nutrients for growth that
includes apple, citrus fruits, corn, potato and banana waste [27].
The Agroindustrial residues are normally considered to be the excellent substrate for the
production of enzyme through SSF technique. The SSF serve as an anchorage for the growing

microbial cultures and also it prevents bacterial contamination due to low moisture and content
of the fermenting medium, the pectinase synthesis is also less affected by the catabolic repression
when compared with SmF. Over all, SSF is much better than SmF for the production of
pectinolytic enzymes using agricultural wastes [28,29,30]
Pectinases are an enzyme group that catalyzes pectic substance degradation through
depolymerization (hydrolases and lyases) and deesterification (esterases) reactions. The wellknown pectinolytic enzymes are homogalacturonan degrading enzymes.

Protopectinases solubilize protopectin forming highly polymerized soluble pectin [5, 9].
They are classified into two types: one reacts with the polygalacturonic acid region of
protopectin, A type; the other with the polysaccharide chains that may connect the
polygalacturonic acid chain and cell wall constituents, B type [12].
Pectin methyl esterase or pectinesterase (EC 3.1.1.11) catalyzes deesterification of the
methoxyl group of pectin forming pectic acid and methanol. The enzyme acts preferentially
on a methyl ester group of galacturonate unit next to a non-esterified galacturonate unit.
It acts before polygalacturonases and pectate lyases which need non-esterified substrates [9].
It is classified into carbohydrate esterase family [13].
Pectin acetyl esterase (EC 3.1.1.-) hydrolyses the acetyl ester of pectin forming pectic
acid and acetate [14]. It is classified into carbohydrate esterase families 1 2and 13 [13].

Polymethylgalacturonase catalyzes the hydrolytic cleavage of -1,4-glycosidic bonds in

pectin backbone, preferentially highly esterified pectin, forming 6-methyl-D-galacturonate
[6].

Polygalacturonase catalyzes hydrolysis of -1,4-glycosidic linkages in polygalacturonic

acid producing D-galacturonate. It is classified into glycosyl-hydrolases family 28[13].
Both groups of hydrolase enzymes (PMG and PG) can act in an endo- or exo- mode. Endo-PG
(EC 3.2.1.15) and endo-PMG catalyze random cleavage of substrate, exo-PG (EC 3.2.1.67) and
exo-PMG catalyze hydrolytic cleavage at substrate nonreducing end producing on a
galacturonate or digalacturonate in some cases [4, 9]. Hydrolases are produced mainly by fungi,
being more active on acid or neutral medium at temperatures between 40 C and 60 C