Global Journal of
Traditional Medicinal Systems
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GJTMS
ISSN- 2319-8761
Global J Trad Med Sys 2014 3(1): 1 - 9
©2012 BRNSS pub hub. All right reserved
Microbial Pectinases: A Review
P. Saranraj1* and M.A. Naidu2
1 - Department of Microbiology, Annamalai University, Chidambaram – 608 002, Tamil Nadu, India.
2 – Department of Pharmaceutics, Mandsur Institute of Pharmacy, Mandsur, Madhya Pradesh, India.
E- mail: microsaranraj@gmail.com
Correspondence to: Dr. P. Saranraj, Department of Microbiology, Annamalai University, Annamalai Nagar – 608 002,
Chidambaram, Tamil Nadu, India.
Received date: 20 – 11 - 13
Accepted date: 05 – 12 - 13
Abstract
Pectinases are the groups of enzymes, which cause degradation of pectin that are chain molecules with a rhamnogalacturonan
backbone, associated with other polymers and carbohydrates. These pectinases have wide applications in fruit juice industry and
wine industry. In fruit juice industry, it is used for clarification, where reduction in viscosity is caused which ultimately leads to
formation of clear juice. They increase the yield of juices by enzymatic liquefaction of pulps; these pectinases also helps in
formation of pulpy products by macerating the organized tissue into suspension of intact cells. In wine industry pectinases are
mainly used for decreasing astringency by solubilizing anthocyanins without leaching out procyadin polyphenols, and pectinases
also increase pigmentation by extracting more anthocyanins. The present review paper contains the following topics: Structure
and properties of pectin, Pectinolytic microorganisms, Pectic enzymes and classification of pectic enzymes, Production of pectic
enzymes, Methods of pectinase enzyme production, Substrates used for pectinase enzyme production, Assay of pectic enzymes,
Nutritional factors affecting enzyme production, Environmental factors influencing enzyme production and Industrial application
of pectic enzymes.
Key words: Pectinases • Pectin • Pectinolytic microorganisms • Fermentation
1. INTRODUCTION
Pectinases are group of enzymes that attack pectin
and depolymerize it by hydrolysis and transelimination as
well as by deesterification reactions, which hydrolyses the
ester bond between carboxyl and methyl groups of pectin.
These enzymes act on pectin, a class of complex
polysaccharides found in the cell wall of higher plants and
cementing material for the cellulose network. Pectinases
accounts for 10% of global industrial enzymes produced and
their market is increasing day by day [1]. Pectinases are
classified according to their mode of secretion as extracellular
and intracellular pectinases. An extracellular enzyme is
excreted (secreted) outside the cell into the medium in which
that cell is living. Extracellular enzymes usually convert large
substrate molecules (i.e. food for the cell or organism) into
smaller molecules that can then be more easily transported
into the cell, whereas an intracellular enzyme operates within
the confines of the cell membrane. Membrane proteins remain
attached in some way to the cell membrane. Both intracellular
and extracellular pectinases are classified on the mode of their
attack on the galacturonan part of pectin molecules [2].
Pectinases can be produced by both submerged and
solid state fermentation (SSF). Submerged fermentation is
cultivation of microorganisms on liquid broth. It requires high
volumes of water, continuous agitation and generates lot of
effluents. SSF incorporates microbial growth and product
formation on or within particles of a solid substrate [3] under
aerobic conditions, in the absence or near absence of free
water, and does not generally require aseptic conditions for
enzyme production [4].
2014|Volume 3|Issue 1|Pg: 1 - 9
Many filamentous fungi like Aspergillus niger,
Aspergillus awamori, Penicillium restrictum, Trichoderma
viride, Mucor piriformis and Yarrowia lipolytica are used in
both submerged as well as solid state fermentation for
production of various industrially important products such as
citric acid, ethanol, etc. Fungi like Aspergillus niger,
Aspergillus oryzae, Penicillium expansum, which are
Generally regarded as safe (GRAS) by United States Food and
Drugs Administration (USFDA) are employed in food
industry. Some bacteria (Bacillus licheniformis, Aeromonas
cavi, Lactobacillus, etc), yeasts like Saccharomyces, Candida
and Actinomycetes like Streptomycetes are also used.
Amongst these, the filamentous fungi are most commonly
employed [5, 6, 7].
Fungi can produce both intracellular as well as
extracellular enzymes. All fungi are hetrotrophic, and rely on
carbon compounds synthesized by other living organisms.
Small molecules like mono disaccharides fatty acids and
amino acids can easily pass through but for breaking down of
larger complex compounds like pectin, fungi secrete extra
cellular enzymes. It is well known that as compared to
intracellular enzymes, the extra cellular enzymes are easier to
be extracted. Intracellular enzymes require more time and
costly chemicals for extraction. Till date, substrates used for
solid - state fermentation are materials of plant origin like
grains such as rice, corn, root, tubers and legumes. Apart from
these, pomace, mango peels, orange waste like peels and other
fruit and vegetable industry waste are also being in much use
[8]
.
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Dr. P. Saranraj and M.A. Naidu /Microbial Pectinases: A Review
Pectinases are frequently used in fruit and vegetable
industry and pectin is also employed widely in food industry.
Peel oil finds many useful applications in both food and
pharmaceutical industry. It is good for the skin citrus solvent
is a biodegradable solvent occurring in nature as the main
component of citrus peel oil. Citrus solvents have pleasant
aroma, & FDAGRAS rating ("generally recognized as safe")
makes it suitable to be used as solvent, citrus solvent can
replace a wide variety of products, including mineral spirits,
methyl ethyl ketone, acetone, toluene, glycol ethers, and of
course fluorinated and chlorinated organic solvents. Dietary
fibers are the most recent value added product and are used as
means of roughage.
2. STRUCTURE AND PROPERTIES OF PECTIN
Pectin is a heteropolysaccharide with galacturonic
acid and methanol as the main components. The pectic
polysaccharide consisted of a traid of polymers: -1, 4. D polygalacturonide, a highly branched L - araban and  -1, 4D-galactan. In addition to D - galacturonic acid, the sugars
such as L - rhamnose, L - arabinose, D - galactose, D -xylose
and L - fructose also present. The carboxylic acid groups of
galacturonic acid residues are partially esterifies with
methanol and methoxyl content varies with source. When all
the carboxyl groups in polygalacturonic acid are esterified, the
methoxyl content is 16.32 per cent i.e. the degree of
esterification is 100 per cent. Acidic and neutral pectins were
carrying ferulic acid on the non reducing ends of the neutral
arabinose and or galactose containing domains. The pectins
carry approximately one teruloyl residue per 60 sugar residue.
These feruloyl pectic acids are involved in the regulation of
cell expansion, in disease resistance and in the initiation of
lignification. The most important unique physical property of
pectins is their ability to form gels with sugar and acids. The
maceration of pectic substances with the help of pectinolytic
microorganisms attribute to the release of bast fibres from the
stem cortex during the retting of flax, jute and other stem fibre
crops, freeing of coffee, cocoa, and white pepper seeds from
surrounding pulp and mucilage. Forgarty and Kelly [9]
reported the presence of pectin in some fruits and vegetables.
3. PECTINOLYTIC MICROORGANISMS
Pectins are degraded by number of microorganisms
to produce a variety of compounds and enzymes which are
involved in many industrial applications. Many pathogenic
bacteria and fungi are capable of degrading pectins [10].
3.1. Pectinolytic Bacteria
Elyrod [11] first reported that the bacterium Erwinia
sp. can degrade pectin. Zucker et al. [12] and Chatterjee et al.
[13]
showed the production of inducible and extracellular
endopolygalacturonase by Pseudomonas fluorescens and
Erwina. Bacteria like Bacillus, Pseudomonas and
Micrococcus isolated from retting flax, jute, sisal and coir and
Erwinia from coffee fruits have shown to possess the ability
to degrade pectin by producing pectinolytic enzymes [14].
Forgarty and Kelly [15] listed many microorganisms
that are capable of degrading pectin. McMillan et al. [16];
Heikinheimo et al. [17]; Weber et al. [18] and Liao et al. [19]
found that many species of Erwinia, Xanthomonas and
Pseudomonas are capable of producing pectinolytic enzymes.
The determination to total counts and total pectinolytic counts
on pulped fruits indicated that pectinolytic bacteria are the
important fraction of the microbial population [20]. The
microbial
flora
of
fruit
peels
consist
mostly
Enterobacteriaceae particularly the genus Erwinia, which
produce pectinolytic enzymes during fermentation.
3.2. Pectinolytic fungi
Many fungal species are capable of degrading pectin
by producing different pectinolytic enzymes. The fungus,
Alternaria sesami produced pectinolytic enzymes viz.,
polygalacturonase transeliminase, pectin transeliminase and
polygalacturonase [21].
Shindia [22] reported that temperature variation during
garbage composting led to corresponding changes in the
distribution of pectin degrading fungi in the compost and the
most common pectinolytic fungi were Aspergillus niger,
Aspergillus flavus, Aspergillus terreus, Penicillium
chrysogenum, Fusarium moniliforme, Alternaria alternata,
Cladosporium cladosporioids and Trichoderma reesei and
these fungi also formed the part of pectinolytic microflora of
coffee fruits.
4. PECTIC ENZYMES AND CLASSIFICATION OF
PECTIC ENZYMES
Pectinase are the group of enzymes that catalyze the
breakdown of pectin containing substances. These enzymes
are produced by plants and microbes and are not synthesized
by animal cells [23]. At present, a majority of commercial
enzymes are obtained by employing fungal cultures.
Pectic enzymes have been classified based on
different criteria [24, 25].
The recent classification of
pectinolytic enzymes was based on the method proposed by
Fogarty and Kelly [26]. Basically, there exist three types of
pectic enzymes viz., pectin esterases, which remove methoxyl
residues from pectin, a range of depolymerizing enzymes
(pectinase) and protopectinase, which solubilizes protopectin
to form pectin [27].
5. PRODUCTION OF PECTIC ENZYMES
During the last decade, world interest on pectic
enzymes has increased because of their industrial value,
especially in food and fermentation industries. Pectic
enzymes account for 10 per cent of the total food enzymes
[28]
. Its use in the production of clarified fruit and vegetable
juices is a common practice in the food industry. The
addition of pectinases to the cloudy juice causes a rapid drop
in the viscosity as well as in flocculation of the micelles and
clear juice can be obtained after filtration. Other industrial
uses of pectic enzymes include the extraction of oil, flavours
and pigments from plant materials and maceration of
vegetables and fruits. Cell free preparation of microbial
pectic enzymes and their ability to macerate plant materials
was reported by Chesson [29].
Pectinolytic enzymes are produced by number of
plants and microorganisms and are not synthesized by
animal cells [30]. Fungal sources do provide the largest
variety of bulk commercial enzymes and have the widest
variety of application, which is well documented by Lowe
[31]
. The high enzyme production was achieved through
strain selection, media development, process development
and scale up programmes [32].
6.
METHODS
PRODUCTION
OF
PECTINASE
ENZYMES
Microbial enzymes are commercially produced either
through submerged fermentation (SmF) or solid substrate
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Dr. P. Saranraj and M.A. Naidu /Microbial Pectinases: A Review
fermentation (SSF) techniques. The SmF techniques for
enzyme production are generally conducted in stirred tank
reactors under aerobic conditions using batch or fed batch
systems. High capital investment and energy costs, and the
infrastructural requirements for large - scale production make
the application of SmF techniques in enzyme production more
impractical in a majority of developing country environments.
Submerged fermentation is cultivation of microorganisms on
liquid broth it requires high volumes of water, continuous
agitation and generates lot of effluents. SSF incorporates
microbial growth and product formation on or within particles
of a solid substrate under aerobic conditions, in the absence or
near absence of free water and does not generally require
aseptic conditions for enzyme production [33]. For the
industrial production of pectinolytic enzymes, it was
important to improve the cultural conditions, yielding better
production of extracellular enzymes in liquid culture on
inexpensive carbon sources [34].
6.1. Submerged fermentation (SmF)
Submerged fermentations (SmF) have the advantages
of better opportunities for process control and analysis and the
basis for planned experiments to increase fermentation yield
through the use of optimized medium [35]. Pectinolytic
enzymes on a large scale are produced from species of the
genus Aspergillus [36] and Penicillium [37].
6.2. Solid state fermentation (SSF)
Solid state fermentation (SSF) is generally defined as
the growth of microorganisms on solid materials in the
absence of free water. It has tremendous potential for the
production of enzymes [38, 39]. Studies on the production of
enzymes by solid state fermentation are increasing because of
the potential advantages such as simplicity, high productivity
and concentrated products over submerged fermentations [40,
41]
.
7.
SUBSTRATES
PRODUCTION
USED
FOR
PECTINASE
Medium require presence of available nutrients and
absence of toxic or inhibitory constituent medium carbon,
nitrogen, inorganic ions and growth factors are also required.
For submerged fermentation, besides carbon source, nitrogen
growth factors media requires plenty of water. The most
widely used substrate for solid state fermentation for pectinase
production are materials of mainly plant origin, which include
starchy materials such as grains, rice, corn, roots, tubers and
legumes, and cellulosic lignin, proteins, and lipid materials
[42]
. Agricultural and food processing wastes such as wheat
bran, cassava, sugar beet pulp, citrus waste, corn cob, banana
waste, saw dust and fruit pomace (apple pomace) are the most
commonly used substrates for SSF for pectinase production
[43]
.
8. MICROORGANISMS INVOLVED IN PECTINASE
PRODUCTION
Microorganisms are currently the primary source of
industrial enzymes: 50% originate from fungi and yeast; 35%
from bacteria, while the remaining 15% are either of plant
origin. Filamentous microorganisms are most widely used in
submerged and solid -state fermentation for pectinase
production. Ability of such microbes to colonize the substrate
by apical growth and penetration gives them a considerable
ecological advantage over non - motile bacteria and yeast,
which are less able to multiply and colonize on low moisture
substrate [44]. Among filamentous fungi, three classes have
gained the most practical importance in SSF; the
phycomycetes such as genera Mucor; the ascomycetes genera
Aspergillus and basidiomycetes especially the white rot fungi
[45]
. Bacteria and yeasts usually grow on solid substrates at the
40% to 70% moisture levels. Common bacteria in use are
Bacillus licheniformis, Aeromonas cavi, Lactobacillus and
common yeasts in use are Saccharomyces and Candida.
Pectinase production by Aspergillus strains has been observed
to be higher in solid state fermentation than in submerged
process [46].
Commercial pectinases are often produced from
fungal sources in liquid broths. Aspergillus and Trichoderma
are widely used for the enzyme production. Pectinase
production has been reported in solid state cultures employing
agricultural by products like cassava fibrous waste [47], wheat
bran [48], apple pomace [49] and citrus wastes [50] as substrates
and these substrates are found to be the best substrates for the
SSF process [51]. Trejo Hernanadez et al. [52] compared the
pectinase yields and productivity by both techniques
suggesting that SSF is more productive than SmF.
Aspergillus niger, a filamentous fungus produces
several pectinolytic enzymes and are currently used in fruit
juice and wine industries [53]. SSF cultures showed higher
pectinolytic activities than those obtained by SmF. In SSF,
exopectinase activity was maximum, after 72 hours while in
SmF, it was delayed further. Pectin lyase production by SmF
peaked after 4 days. The comparative ratios of productivities
(SSF/SmF) obtained for endo - exopectinase and pectate lyase
were 6.51 and 29 respectively showing that, overall the SSF
technique is more productive than SmF [54] and the results
confirmed the findings of earlier study on pectinase
production using the strain Aspergillus niger [55].
Cotty et al. [56] determined the pectinases production
by Aspergillus flavus by measuring the clear zones formed
around the colonies stained with ruthenium red. Out of 87
isolates tested, 15 produced red zone after staining with dye.
Major achievement of this study was to screen out most
efficient pectinases producing isolates of Aspergillus niger
from the local environment to fulfill the requirement of local
demand.
Mehta et al. [57] screened one hundred sixty eight
bacterial strains, isolated from soil and samples of vegetable
in decomposition for the use of citrus pectin as the sole carbon
source. Out of 102, 168 were positive for pectinase
depolymerization in assay plates as evidenced by clear
hydrolization halos. Among them, 30% presented
considerable pectinolytic activity. The cultivation of these
strains by submerged and semi - solid fermentation for
polygalacturonase production indicated that five strains of
Bacillus sp. produced high quantities of the enzyme. The
physico-chemical characteristics, such as optimum pH of 6.0
to 7.0, optimum temperatures between 45°C and 55ºC,
stability at temperatures above 40ºC and in neutral and
alkaline pH, were determined.
Urmila Phutela et al. [58] isolated thermophilic fungal
strain producing both pectinase and polygalacturonase after
primary screening of 120 different isolates. The fungus was
identified as Aspergillus fumigatus. Using solid state
cultivation, the optimum levels of variables for pectinase and
polygalacturonase (PG) production were determined.
Maximal levels of enzyme activities were achieved upon
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Dr. P. Saranraj and M.A. Naidu /Microbial Pectinases: A Review
growing the culture in a medium containing wheat bran,
sucrose, yeast extract and (NH4)2SO4 after 2 - 3 days of
incubation at a temperature of 50ºC. Highest enzyme activities
of 1116 Ug-1 for pectinase and 1270 Ug-1 for
polygalacturonase were obtained at pH 4.0 and 5.0,
respectively.
9.2. Lyase enzyme
Enzyme activity of lyases can be measured by
following the increased absorption of the digest at 560 nm
wavelengths. Because of its simplicity, the assay procedure is
widely and routinely used to assay polygalacturonate lyase
and polymethyl galacturonate lyase [68].
Chawanit Sittidilokratna et al. [59] carried out the
screening of pectinase producing bacteria and assessment of
the effectiveness for biopulping of paper mulberry bark of the
pectinase of the highest producer. Pectinolytic bacteria were
initially screened from 6 identified and 118 unknown isolates.
Twelve strains gave positive results, including 3 of Erwinia
carotovora subsp. carotovora, 2 of Erwinia chrysanthemi and
7 of Bacillus sp. Crude pectinases were prepared from the
selected strains. Then, the activity of 3 pectinase types,
namely polygalacturonase (PG), pectate lyase (PAL) and
pectin lyase (PL) was investigated. The results showed the
highest PG production from Erwinia chrysanthemi strain N05
isolated from onion and highest PAL and PL production from
Bacillus sp. strain N10 isolated from paper mulberry bark.
Both, N05 and N10 possess similar optimum conditions at pH
10.0 and 35ºC, and were stable at pH 3 - 12 for 30 minutes
and 20 - 40ºC for 24 hours.
10. NUTRITIONAL FACTORS AFFECTING ENZYME
PRODUCTION
Abdul Hannan et al. [60] performed a multistep
evaluation of 52 strains of Aspergillus niger on the basis of
polygalacturonase production. Plate method was employed for
preliminary screening of isolates. The strains exhibiting
relative clear zone with diameter above 1.386 were selected
for final screening. Out of 52 strains, 23 were selected for
further screening by using optimized enzyme assay method.
The enzyme production by 23 selected strains was studied in
submerged fermentation with pectin as the only carbon
source. Maximum activity was observed in two isolates of
Aspergillus niger (H12 and H51) showing more than 0.700
units after 96 hrs incubation period. Only four isolates (H06,
H13, H45, and H46) gave the maximum activities after
incubation for 72 hours whereas decline was observed at 96
hrs.
Lali Kutateladze et al. [61] investigated the physiology
and some biochemical characteristics of the selected strains.
The nutrient media for each particular strain was optimized
and conditions of growth were established. The strain
Penicillium canescens I-85 reveals the highest pectinase
activity at 27ºC and pH 4.0; the strain Aspergillus niger at
40ºC, pH 6; Trichoderma viride Ts-2 at 30ºC, pH 7.5. As a
result of optimization of the Nutrient media the activity of
pectinase increased by 122, 28 and 98%, respectively.
9. ASSAY OF PECTIC ENZYMES
Assay method for the detection and measurement of
pectinolytic activity ranged from pure qualitative methods [62]
for demonstrating the presence of enzymes activity to
quantitative methods [63, 64], which determine the activity in
terms of actual linkages hydrolyzed.
9.1. Pectic esterase
The most common method for determining pectin
esterase activity is the titrimetric method of estimation of
carboxyl groups formed in pectin by the enzyme [65, 66]. Pectin
esterase can also be assayed by measuring the methanol
liberated from pectin during the reaction. Zhao et al. [67] have
described a simple spectrometric method for the estimation of
methanol released from pectin.
Tuttobello and Mill [69] reported that the high enzyme
yields were obtained in a medium of groundnut flour with 2
per cent sucrose and 2 per cent pectin. Moran and Starr [70]
reported that Erwinia carotovora and Erwinia aroideae were
constitutive with respect to endopolygalacturonate lyase
synthesis. In batch fermentation, the differential rate of
enzyme formation was low with glucose, medium with
glycerol and high with pectate as the sole carbon and energy
source and it was concluded that enzymes production is under
catabolism repression control.
The use of complex substances like citrus peel or
other agricultural wastes is common, since these materials are
natural substrates for pectinolytic enzymes [71, 72]. These
substance and their degradation products are inducers for the
enzyme synthesis, e.g., Galacturonic acid and pectic acid [73]
as well as polygalacturonic acid and pectin itself [74].
Ward and Fogarty [75] stated that glucose is the most
suitable carbon source for the production of the
polygalacturonate lyase from Bacillus subtilis and sodium
polypectate was the best inducer of polygalacturonate lyase
with Flavobacterium pectinovorum.
Bateman [76] showed that the pectic acid or sodium
polypectate was found to be a better substrate for the
polygalacturonase production than pectin. Kunte and Shastri
[77]
reported that the maximum production of
polygalacturonase and polymethgalacturonasel took place in a
medium containing 0.5 per cent pectin and 0.5 per cent
cellulose powder. Maximum production of PG occurred after
4 days and the optimum incubation period for the production
of pectinolytic enzymes vary from strain to strain and species
to species.
Foda et al. [78] reported that for maximum
pectinolytic enzyme production, Asperillus aculeatus and
Mucor pusillu fungi require pectin as carbon source. Murad
and Foda [79] found that supplementation of permeate with
yeast extract or peptone resulted in marked increase in the
enzyme activity as compared to the control permeate medium.
Garzon and Hours [80] stated that by using Aspergillus
foetidus, pectinase with an activity of 1600 - 1700 Ug-1 after
36 hrs of culture can be obtained from citrus waste
supplemented with yeast extract and mineral salts.
Pectinase produced by SmF with Aspergillus and
Fusarium was induced by pectin and its derivatives [81]. Sara
et al. [82] reported that under solid state fermentation exo and
endo pectinase activities of Aspergillus niger increased with
increase in the concentration of carbon source. She also found
that under SmF, the exo and endo pectinase activities by
Aspergillus niger sharply decreased when glucose or sucrose
(3 per cent) was added to the pectin containing culture
medium. Bahkali [83] reported that the fungus Verticillium
tricorpus exhibited maximum activity of polygalcturonase
enzyme in medium containing pectin as substrate. He reported
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Dr. P. Saranraj and M.A. Naidu /Microbial Pectinases: A Review
that pectin is the best substrate for the production of
polygalacturonase.
12. INDUSTRIAL
ENZYMES
Gupta et al. [84] reported that sugar beet shreds and
apple pomace produced higher polygalacturonase activity
under semi solid conditions and the raw onion and citrus peel
proved better under stationary condition. Isshiki et al. [85]
reported that Alternaria alternata (AG 325) produced
polygalacturonase in liquid medium containing one percent
pectin. Increased synthesis of polygalactuonase was also
recorded with the addition of five percent of sucrose to the
culture medium.
Application of enzymes in biotechnological process
has expanded considerably in recent years. In food and
related industry, major importance was being attached to the
use of enzymes in upgrading quality, increasing yields of
extractive processes, product stabilization, and improvement
of flavour and by product utilization [95].
11. ENVIRONMENTAL FACTORS INFLUENCING
ENZYME PRODUCTION
In order to optimize enzyme production, parameters
affecting the enzyme synthesis have to be standardized.
Although, optimum conditions may vary for each organism
and enzyme certain factors have been established as the most
significant in influencing overall enzyme yield [86]. The
critical factor for fungal growth on solid surface is moisture.
The control of moisture level within a relatively narrow range
was essential for optimizing solid state fermentation. Many
microorganisms can grow in solid substrate but only
filamentous fungi can grow in the absence of free water. A
properly moistened substrate would have a surface film of
water to facilitate dissolution and mass transfer of nutrients
and oxygen, but antiparticle channels would be left free to
permit oxygen diffusion and heat dissipation [87].
Norkrans and Hammarstrom [88] found that Rihizinia
maculata produced polygalacturonase optimally at a pH range
of 3 - 4 and the activity dropped to 50 percent at the pH range
5 - 6. Foda et al. [89] reported that the optimum pH value for
enzyme production were 4 - 5 and 4 - 6 for Aspergillus
aculeatus and Mucor pusillus, respectively and both these
organisms require pectin as carbon source. Moran and Starr
[90]
indicated that polygalacturonase secretion was maximum
at pH 4 - 5 in the case of Fusarium oxysporum. Hours et al.
[91]
found the highest pectinolytic activity from apple pomace
at pH 4.0 and maximum saccharification of sugar beet pulp
was found at pH 4.8 [92]. The biggest obstacle to scale up the
SSF process is heat build-up. This causes evaporative water
loss and stops vegetative growth. On the other hand, a
controlled evaporation with continuous water replacement
may promote heat dissipation and assures productive
vegetative growth.
Pectinolytic activities of Aspergillus niger produced
in SSF are more stable than those produced in SmF.
Maximum endo polygalacturonase activity in SSF was
reported at 60C [93]. They also reported that exopectinase
activity obtained in SmF. According to Pandey [94], the major
factors that affect the microbial synthesis of enzyme in solid
state fermentation include; selection of a suitable substrate
and microorganisms, pre - treatment of the substrate, particle
size (inter particle space and surface area) of the substrate,
water content and water activity (aw) of the substrate, relative,
humidity, type and size of the inoculum, control of
temperature of fermenting matter, removal of metabolic heat,
period of cultivation, maintenance of informality in the
environment of solid state fermentation system and the
gaseous atmosphere.
APPLICATION
OF
PECTIC
In juice extraction, both maceration and viscosity
reduction contribute to the increased yields of press juice
obtained. It is the ability of pectic enzyme to reduce viscosity
of drinks from fruits [96]. Fogarty and Kelly [97] reported the
use of pectinases in wine clarification. Hours et al. [98]
reported that pectinase levels ranging from 1000 - 2000 U l-1
of juice for 1 to 3 hours are necessary to achieve clarification
in three different apple juices.
Pectolytic enzymes are added before fermentation of
white wine musts, which are made from pressed juice without
any skin contact in order to hasten clarification. Another
application of pectolytic enzymes during wine making was
associated with the technology of thermovinification. During
heating the grape mash for few hours large amounts of pectin
are released from the grape, this does not occur in traditional
processing. It is therefore necessary to add a pectolytic
preparation of the heated mash, so that the juice viscosity is
reduced. An additional benefit from the process is that the
extraction of anthocyanins was enhanced, probably due to a
breakdown in cell structure by the enzyme, which allows the
pigments to escape more readily and thus helps in color
enhancement [99].
In the textile industry pectinases are sometimes used
in the treatment of natural fibers such as linen and ramie fibers
[100]
. Das and Baruah [101] reported that saprophytic fungi
Trichoderma reesei isolated from arecanut husk secreted high
polygalacturonase and when they treated cell free preparations
of polygalacturonase enzyme on arecanut seeds, the
germination was better than those of seed nuts treated with
distilled water. Marcia Soares et al. [102] stated that pectinases
were used in the processing of green coffee beans to hasten
the removal of the jelly that surrounds the coffee cherry.
Natural fermentation may give a coffee bean of inferior
quality.
One of the potential applications of pectic enzymes
involves the treatment of commercial softwoods. The age old
practice of retting by which many important textile fibres such
as flax, hemp and jute are prepared involves the pectinolytic
enzymes of certain microorganism. Pectinolytic enzymes may
also be applied in disrupting gels in order to assist recovery of
oils [103].
Pectinolytic enzymes play a crucial role by
increasing the access of cellulases to their substrates [104, 105].
Wang and Chang [106] and Spagnulo et al. [107] reported that
pectinase appeared to be the most important enzyme, since by
hydrolyzing the pectic surface of the lignocellulosic
substrates; it favored the degradation of cellulose and
hemicelluloses by the respective enzymes.
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