Journal of Mines, Metals and Fuels, 71(1): 408-417; 2023. DOI: 10.

18311/jmmf/2023/33750

Print ISSN : 0022-2755

Journal of Mines, Metals and Fuels

Contents available at: www.informaticsjournals.com/index.php/jmmf

Microbial Production Strategies for the Production

of Industrially Important Pectinases: Bioreactor
Consideration and its Applications in Various Sectors
V Vaishnavi1, G Divyashri2*, Thomson Tanya1, S Sejal1 and T P Krishnamurthy1
1Department of Biotechnology, M S Ramaiah Institute of Technology, Bangalore 560 054, India.
2*Assistant
Professor, Department of Biotechnology, M S Ramaiah Institute of Technology. E-mail: gdivyashri@gmail.com;
gdivyashri@msrit.edu

Abstract
Pectinases are a group of enzymes hydrolyzing pectin substances. Pectin esterases (catalyze the removal of methoxyl residues
of pectin), depolymerizing pectinases and protopectinases (converts insoluble protopectin into soluble pectin) are the three
major types of pectinases, which are of commercial importance. Pectinases are abundant in nature and are synthesized by
a variety of microorganisms, notably from bacteria, fungus to yeast. Production of pectinases by various microorganisms is
through solid state and submerged fermentation procedures. Optimization of essential bioreactor considerations and
parameters such as temperature, pH and fermentation times play a vital role in pectinase production. The need for
commercial production of microbial pectinase has expanded globally due to its applications in various sectors. Pectinases
are being used extensively in the food, agricultural, environmental and pharmaceutical sectors. Hence, it needs to be
explored further so as to ensure its maximum usage in various industries. This review discusses the structure of pectin and
pectinase, substrate specificity, mode of action, microbial production strategies and its industrial applications. It also
discusses the current limitations and future prospective.

Keywords: Pectinases, Pectin esterases, Protopectinases, Solid-state fermentation, Submerged fermentation, Industrial
applications

1.0 Introduction some plant tissues during maturation and storage 49 .

1.1 Pectinase
The bioactive substances known as enzymes control a
variety of chemical alterations in living tissues. Pectinases are
a collection of interconnected enzymes responsible the
hydrolysis of pectin compounds, which are primarily found
in plants. Higher plants and microbes both have pectinolytic
enzymes in large quantities. They are crucial for plants
because they aid in cell wall extension and the softening of
*Author for correspondence
According to the cleavage site, pectinases are divided into
three groups:
(1) hydrolases consisting of polygalacturonase, PG (EC
3.2.1.15)
(2) lyase/trans-eliminases comprising pectinlyase, PNL
(EC 4.2.2.10), and pectate lyase, PL (EC 4.2.2.2);
(3) pectin esterase, PE (EC 3.1.1.11)67.
In order to accelerate the extraction of fruit juice from fruit,
such as apples and sapota, pectinase enzymes are frequently
utilised in processes involving the breakdown of plant
materials. Since the 1960s, pectinases have also been utilised
in the manufacture of wine53.

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 Microbial Production Strategies for the Production of Industrially Important Pectinases: Bioreactor Consideration and its Applications in Various Sectors
Figure 1: Mode of action and byproducts (a) PG (polygalacturonase) and PMG (polymethylgalacuronases) (b) PE (pectin esterase),
PGL (polygalacturonate lyase) (c) PGL (Polygalacturonate lyase) and PL (Pectin lyase)
1.2 Mode of Action
Based on mode of action, the pectinase enzyme can be
roughly divided into three groups (Fig.1): pectin esterase,
hydrolases, and lyases. The methoxyl group of pectin is de-
esterified by pectin esterase, resulting in the formation of
pectic acid. Hydrolases (Polygalacturonases and Polymethyl-
galacturonases) catalyse the hydrolytic cleavage of a-1,4-
glycosidic linkage in pectic acid and pectin, respectively, while
lyases (Polygalacturonate Lyase and Polymethyl-
galacturonate Lyase) catalyse the cleavage of a-1, 4-glycosidic
linkage by trans-elimination and forming unsaturated
galacturonates and methyl galacturonates respectively.
1.3 Substrate for Pectinase
The plant cell wall, is a structure that encloses and
safeguards the cell and is unique to plants and necessary to
their life, is what sets them apart from other living things.
Plants’ mobility is restricted as a result, which makes them
plastic and gives them the ability to endure various adverse
environmental conditions, as well as to withstand attack by
pathogens and herbivores. The composition of the cell wall,
which is made up of polysaccharides, proteins, aromatic, and
aliphatic chemicals allows for the growth of plants in a variety
of environmental settings. As a cell develops and the
environment changes, cell wall structure is continuously
adjusted. During the first stages of cell expansion and growth,
the plant cell develops the middle lamella and the primary wall.
Many cells have an additional secondary wall that thickens
and strengthens the primary wall. According to11, the primary
wall may help with signal transduction, cell adhesion, and wall
structural integrity.
Cellulose, hemicellulose, and pectic compounds make up
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the cell walls of immature plants. Hemicelluloses and pectic
materials serve as the cellulose network’s reinforcement, while
the cellulose microfibrils give the cell wall its strength. The
integrity and rigidity of plant tissue are influenced by pectins
or pectic compounds, which are involved in intricate
physiological processes like cell development and
differentiation44. The primary walls of most plant tissues are
composed of pectin.
Pectins are a class of galacturonic acid-rich plant cell wall
polysaccharides that are covalently bonded2. About 70% of
pectin is made up of galacturonic acid, and all of the pectic
polysaccharides have it connected at the O-1 and O-4
positions40.
1.4 Substrate Specificity
Analysis of substrate specificity showed that the
pectinase enzyme is capable of acting on a variety of
substrates. Thermo-alkaline pectinase from Hylocereus
polyrhizus can act on polysaccharides such as arabinan,
wheat arabinoxylan and oat spelt xylan , according to a study
on the enzyme’s substrate activity4. Based on a study on
pectinase produced by Aspergillus niger LFP-1 on specific
substrate (peels of Citrus maxima)25, it was discovered that
pectin served as the primary substrate for the pectinase
enzyme. This discovery agreed with research by 12 on
pectinase released by Rhizopus oryzae and it concluded that
the enzyme exhibits significant activity against a fruit-based
substrate. In addition, the enzyme had up to 20% relative
activity in the degradation of oat and polymeric carbohydrate
(starch). The pectinase did prove ineffective against a number
of substrates, including cornflakes, dextrin, and locust bean
gum (LBG). The earlier investigations, however, used a variety
of substrates to determine the specificity of pectinase’s
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V Vaishnavi, G Divyashri, Thomson Tanya, S Sejal and T P Krishnamurthy
substrate. In the substrate specificity experiment,
polygalacturonase acid was utilised as a substrate, and55
reported the production of 8.34 U/mL of pectinase (100%
relative activity). Additionally, digalacturonic acid was utilised
as a substrate in another study to examine the enzyme
specificity of Streptomyces lydicus pectinase, reaching 56%
of the maximum enzyme concentration24.
2.0 Microbial Production Strategies
2.1 Microorganisms Producing Pectinases
Industrially, microorganisms are preferred as the source
for the production of pectinases instead of plants or animals
due to various advantages. They provide for easy, consistent
and cost-effective production. Numerous microorganisms can
release pectinases, including bacteria27, yeast47, and fungi69.
Aspergillus niger, the primary producing strain, is a
filamentous fungus that produces a majority of the microbial
pectinolytic enzyme used in industry 64. These
microorganisms use pectin as a carbon source and degrade
the pectin substances to produce pectinolytic enzymes.
2.1.1 Pectinase producing bacteria
Erwinia species, Pseudomonas fluorescens,
Pseudomonas, Bacillus licheniformis, Aeromonas cavi,
Lactobacillus and Micrococcus have a high capacity to
breakdown pectin. From a study done on the estimation of
pectinolytic activity of pectinase producing bacteria, two
isolates were identified as Bacillus and Pseudomonas.
Significant amounts of pectin degrading enzymes, including
pectin esterase and pectate lyase, were detected in all of the
bacterial cultures. The bacterial isolate Pseudomonas sp.
displayed higher levels of pectinase activity. Additionally,
according to Chatterjee et al., pectinases could be produced
by a number of bacteria, including Erwinia sp., Pseudomonas
sp., and Bacillus sp. Based on the activity of such pectinases,
the isolated cultures were evaluated for further studies.
Pseudomonas sp., and Bacillus sp., two of the bacterial
isolates, had the highest pectinase activity18.
Alkalophilic bacteria such as Bacillus spp. are mainly
responsible for producing alkaline pectinases. Bacillus sp.
DT7, a soil isolate, was shown to produce substantial
amounts of pectin lyase. Bacillus sp. DT7 produced more
pectin lyase due to better development circumstances than
was previously documented in the literature. Using gel
filtration and ion exchange chromatography, this enzyme was
purified. The purifed enzyme worked at its best when the pH
was 8.0 and the temperature was 60C. CaCl2 and
mercaptoethanol considerably improved the pectin-degrading
activity of the isolated enzyme. The textile industry, plant
tissue maceration, and wastewater treatment all benefit greatly
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from the use of this pectinase.
According to a 1942 research by Elyrod, pectin may be
broken down by the bacterium Erwinia sp. with the help of
pectinases. Phytopathogens such as the soft-rotting Erwinia
species, E. carotovora and E. chrysanthemi, produce a group
of pectinolytic enzymes. Bacillus, Pseudomonas, and
Micrococcus have shown to have the ability to produce
pectinases, according to Chessen et al., 1980. Pseudomonas
and Streptomyces species have been identified as other
bacterial genera with pectinolytic characteristics8.
2.1.2 Pectinase producing fungi
Fungi are capable of producing both intracellular and
extracellular enzymes. Fungi, being heterotrophic, rely on
carbon sources produced by other organisms. Despite the
fact that minor biomolecules can easily pass, extracellular
enzymes are secreted by fungi to break down larger
polysaccharides such as pectin. When it comes to
downstream processing, extracellular enzymes are easier to
extract than intracellular ones. Extraction of intracellular
enzymes requires a lot of time and expensive reagents.
Materials of plant origin are generally used as substrates for
solid state fermentation. The substrates include grains like
rice and legumes, tubers and agro-waste substrates such as
pomace, citrus peels, mango peels and other waste materials.
Filamentous fungi like Aspergillus niger, Aspergillus awamori,
Penicillium restrictum, Trichoderma viride, Mucor piriformis
and Yarrowia lipolytica have been employed for the
production of industrially significant enzymes, such as
pectinases (Dalboge, 1997). They are used in SmF and SSF
for the production of various commercially important
products such as ethanol, citric acid etc. Fungi which are
regarded as safe by the USFDA are utilised in the food sector.
Fungal isolates such as Aspergillus niger, Aspergillus oryzae,
Aspergillus flavus and Penicillium chrysogenum are known
to produce pectinolytic enzymes for the breakdown of pectin.
Isshiki et al. and Kapat et al. both confirmed that fungi like
Alternaria, Cladosporium, Colletotrichum, Mucor, Penicillium,
and Trichoderma produce pectinases. Among these,
Aspergillus niger was found to be the most commonly used
species. Different researchers came to different conclusions
about the isolation, characterization, selection,
characteristics, and fermentation of Aspergillus niger strains
for the synthesis of pectinases utilising various substrates.
There have also been reports of pectinase production by
other Aspergillus species, including16. A. giganteus was the
first species to be reported to produce endo-PGL 45 .
Additionally, the generation of pectinase involves species
such as Penicillium, Fusarium, Mucor, Neurospora crassa,
Sclerotinia sclerotiorum, and others. In order to absorb
nutrients from plant tissues and disrupt the middle lamella in
plants, the fungus creates these enzymes.
Journal of Mines, Metals and Fuels
 Microbial Production Strategies for the Production of Industrially Important Pectinases: Bioreactor Consideration and its Applications in Various Sectors
2.1.3 Pectinase producing yeast
The production of pectinolytic enzymes by yeasts is not
preferred as it is seen only in a few species of yeast9. Luh
and Phaff initially documented the endo-PG synthesis by
yeasts in 1951 with Saccharomyces fragilis. They discovered
that some yeast species – Saccharomyces fragilis,
Saccharomyces fragilis var. no. 351, Torulopsis kefyr, Candida
pseudotropicalis and Candida pseudotropicalis var. lactosa,
– represent all yeast genera and were capable of significantly
altering pectin34. The potential of other yeast species to
produce pectinase is also described in recent papers. These
yeast species include Saccharomyces sp., Cryptococcus sp.,
Rhodotorula dairenensis, Aureobasidium pullulans,
Wickerhanomyces anomalus, Geotrichum klebahnii,
Kluyveromyces marxianus, etc8.
2.2 Fermentation Routes (Solid State and
Submerged)
The production of microbial pectinase involves the
following steps - isolation, selection and screening of the
microbial strain capable of producing pectinases, growth of
microorganism on a suitable culture medium, fermentation
and purification methods. The production of pectinase can
be done either using Solid State fermentation or by
Submerged fermentation. SmF involves the growth of
microbes of interest in a liquid medium. Submerged
fermentation (SmF) is the process by which microbes grow
and decompose substrates in the presence of a large amount
of free water (liquid medium), continuous agitation and
generates a large number of effluents41. The method of choice
for industrial activities is SmF6. In solid state fermentation
(SSF), due to absence of free water, microorganisms ferment
the solid substrate33. SSF is often performed in an aerobic
environment. Aseptic conditions are not a prerequisite.
2.2.1 Solid State fermentation
SSF has been used to produce several enzymes such as
xylanses and pectinases by A. awamori using grape pomace
as the substrate. Production of polygalacturonase and pectin
lyase was also done by Penicillium viridicatum using other
fruit-based substrates57, and production of very high levels
of alkaline and thermophilic pectinase by Bacillus sp. DT729
are also relevant examples of the production of pectinolytic
are also relevant examples.
2.2.2 Submerged fermentation
Pectinase can be produced through submerged
fermentation routes too. Production of xylano-pectinolytic
enzymes from Bacillus pumilus was achieved through this
method30. Bacillus licheniformis grown on a medium containing
NaNO3, KH2PO4, KCl, MgSO4, Tryptone and Orange peel7 and
and Aspergillus niger can also be used for the production of
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pectinase through submerged fermentation. Aspergillus niger
can also be used for pectinase production through SmF.
On the fundamental differences between the generation
of pectinase by submerged fermentation (SmF) and solid-
state fermentation, comparative investigations have been
done (SSF). These studies have compared productivity and
enzyme yields of both the techniques and concluded that
solid state fermentation was more productive than submerged
fermentation. This might be due to the fact that SSF is simple,
and produces concentrated products with high productivity1.
2.3 Bioreactor Considerations
From an experiment done by16 on the production of
pectinase in a packed bed reactor, it was observed that the
maximum pectinase yield was seen with specific parameters.
Alternating between 24 and 32 celsius using saturated inlet
air proved optimum. The best result was obtained with bed
height of 40 cm containing 27kg of wheat bran and 3kg of
sugarcane bagasse substrate. Another study by15 included
statistical approaches to seek novel and optimum
compositions of medium components that affect the
production of pectinase. included statistical approaches to
seek novel and optimum compositions of medium
components that affect the production of pectinase. The
study results showed that pectin, (NH4)2SO4 and K2HPO4
affected pectinase production the most. The yield was
increased 2.8 folds in optimized medium when compared to
un-optimized medium. Box-Behnken design was performed to
optimize the concentrations of these components. The
maximum pectinase activity was obtained utilising the middle
levels of all three tested components. A new strain,
Aspergillus niger NRC1ami, which was isolated from citrus
fruit was cultivated on the statistically optimized media.
A.niger requires a continuous supply of oxygen and
controlled pH for the maximum growth and production of
metabolites. It was also noted that scaling up of the cultivator
from shake flasks to bioreactos resulted in an increase in the
yield of pecinase and also other enzymes such as amylase
and invertase. This coupled with pH controlled conditions
increased the production of enzyme about 4-folds. The
growing cells have a considerably more stable growing
environment thanks to pH regulation, which is evident in the
enhanced cell development and productivity. The relationship
between enzyme denaturation and the instability of
conformational configurations and the influence of pH change
on enzyme activity has been shown.The study results
showed that pectin, (NH 4)2SO 4 and K 2HPO 4 affected
pectinase production the most. The yield was increased 2.8
folds in optimized medium when compared to un-optimized
medium. Box-Behnken design was performed to optimize the
concentrations of these components. The maximum pectinase
activity was obtained utilising the middle levels of all three
Journal of Mines, Metals and Fuels | 411
V Vaishnavi, G Divyashri, Thomson Tanya, S Sejal and T P Krishnamurthy
tested components. A new strain, Aspergillus niger NRC1ami,
which was isolated from citrus fruit was cultivated on the
statistically optimized media. A.niger requires a continuous
supply of oxygen and controlled pH for the maximum growth
and production of metabolites. It was also noted that scaling
up of the cultivator from shake flasks to bioreactos resulted
in an increase in the yield of pecinase and also other enzymes
such as amylase and invertase. This coupled with pH-
controlled conditions increased the production of enzyme
about 4-folds. The growing cells have a considerably more
stable growing environment thanks to pH regulation, which
is evident in the enhanced cell development and productivity.
The relationship between enzyme denaturation and the
instability of conformational configurations and the influence
of pH change on enzyme activity has been shown.
3.0 Industrial Applications of
Pectinases
3.1 Applications In Food Industry
3.1.1 Vegetable and fruit processing
A significant step in the processing of fruits and vegetables
is the use of suitable enzymes to treat the pulp32. Consumers
tend to prefer juices that are high in nutrition, have high clarity
but low viscosity. The clarification of juices is generally
achieved by liquefaction of the pulps, removal of peels35 and
maceration of vegetables by pectinases. The degradation of
pectin depends on various factors such as time taken,
composition of the juice and the enzymes used. The treatment
time ranges from 5 minutes to 6 hours (Aliaa et al.2010), pH can
range from 2.5-613, temperature is maintained below 50°C16. In
the process of making apple juice, pectic enzymes are employed
to speed up juice extraction or pressing as well as to help
separate precipitate by sedimentation, filtering, or
centrifugation. Rhamnogalacturonase, a novel pectinase, and
its function in the apple peel lysis were described by52. Initially
identified in Aspergillus aculeatus, this enzyme now appears
to be generated by additional Aspergillus species. Depending
on the specific type of pectinase used, how much is utilised,
the reaction temperature, and the apple variety employed,
pectinase treatment can take anywhere between 15 minutes and
two hours. The production of grape juice is increased by
pectinase enzyme treatment by up to 30%. Juice extraction rises
as pectinase levels increased from 0.05 to 1.5%59.
3.1.2 Wine processing
Pectinolytic enzymes are used in the extraction process
of winemaking to increase yield, enable filtration, and improve
colour and flavour. Wine processed enzymatically showed
greater stability than control wines before the addition of
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inoculum. Clarification of the substrate is necessary before
fermentation can begin; this improves the sensory
characteristics of white wine. Study by Bosso (1993) reported
that the concentrations of 2-phenyl ethanol and iso-amyl
alcohol increased while those of n-propanol decreased in
fermented grape that had been pre-treated with pectic
enzymes. They investigated the combined impact of pectinase
treatment and yeast culture fermentation on ethanol
generation. When treated with pectinase, fermentation was
conducted for 12 hours at 30°C and pH 4.5, increasing the
volume of juice. According to numerous studies, the activity
of pectin esterase causes wine to have higher quantities of
methanol when pectinolytic enzymes are added during the
wine-making process. The maximum amount of methanol that
can be present in wine should be restricted because it is
hazardous. Hence, pectin esterase levels in commercial
combinations should be limited16.
3.1.3 Coffee fermentation
Fermentation of coffee is essential for the removal of
mucilage from parchment coffee. The mucilage is composed of
polysaccharide (pectin), cellulose and starch. The mucilage is
responsible for prolonging the duration required to dry the
coffee brands and may even lead to mold formation. Hence, the
fermentation process is aided my microorganisms present in the
environment or occur naturally in the coffee fruit, that are
capable of producing pectinases. Microorganisms aid by
breaking down the mucilage by releasing enzymes. Recent
studies indicate that fermentation is being used to produce
specialty coffee. According to Vaast et al., a number of
physiological changes take place during this process, including
a reduction in the amount of water and simple sugars and
development of fragrance and flavour precursors. More than
50 yeast and bacterial species have been found in previous
publications to be active during this process. It is claimed that
when pectinaceous carbohydrates are fermented 57, the
microorganisms engaged in coffee fermentation contribute to
making ethanol and other higher carboxylic acids. According
to Coughlan and Mayer, several extracellular enzymes
produced by Bacillus species may help breakdown pectin
compounds found in the peel, pulp, and mucilage of coffee
cherries. The three key enzymes that microorganisms create are
PL, PG, and PME. Unsaturated galacturonic acids are produced
when pectin is broken down by trans-elimination, a process
catalysed by pectin lyase. The primary enzyme involved in the
fermentation of coffee is polygalacturonase. In order to
produce pectic acid (polygalacturonic acid), it catalyses the
hydrolysis of 1,4-glycosidic bonds. The methoxyl group of
pectin is de-esterified by pectin methyl esterase, resulting in
the formation of pectic acid and methanol. Saccharomyces
cerevisiae, Candida parapsilosis, Pichia guilliermondi, and
Pichia fermentans have demonstrated pectinolytic action,
Journal of Mines, Metals and Fuels
 Microbial Production Strategies for the Production of Industrially Important Pectinases: Bioreactor Consideration and its Applications in Various Sectors
resulting in the creation of premium coffee drinks with a
distinctive flavor of caramel and fruits, and intense perceptions
of buttery, and fermented flavors19.
3.2 Applications In Agriculture Sector
3.2.1 Plant fibre retting and degumming
For its oil-bearing seeds, flax is farmed both in tropical and
temperate regions. The remaining cake is used as feed, and
the seeds are crushed to produce linseed oil. Flax retting is a
procedure used to rot away or dissolve the pectin and cellular
tissues that surround fibre bundles, making it easier to
separate the fibre from the stem. One of the most
environmentally friendly processes is microbial retting. The
retting water in commercial retting facilities is a rich source of
microbes that contribute to the retting of flax. Three effective
strains capable of producing pectinase, which is required for
flax retting, were chosen to investigate their involvement in
optimizing the retting process. These strains, which have
specific pectinase activity of 37.17, 33.53, and 28 Umg1
respectively, are Bacillus humi, Chryseobacterium culicis, and
Micrococcus luteus. These strains’ impact on retting time and
fibre quality was researched by {reference}.
Chryseobacterium culicis, Micrococcus luteus, and Bacillus
humi were used in a combination treatment that reduced
retting time by 30%. Chryseobacterium culicis and
Micrococcus luteus were used in a mixed inocula that reduced
retting time by 25%. The retting period was 20% shorter when
Micrococcus luteus and Bacillus humi were combined. After
microbial retting, the measurement of weight loss, tensile
strength, and the whiteness and yellowness of the fibres was
kept track of. The findings suggested that the acquired
particular bacterial strains improved the retting procedure38.
3.2.2 Oil extraction
Pectinases have been used to boost the extraction of
olive oil and enhance its organoleptic qualities, such as
phenolic content. The enzymes are therefore especially
pertinent to the olive oil industry. According to a study by
the most productive strain of pectinase was Aspergillus
giganteus. It was investigated whether the pectinase from A.
giganteus could enhance the extraction of olive oil. When
A. giganteus extract was added at the start of the malaxation,
the number of oil droplets increased, and indicating
improved oil release and malaxation capabilities. The primary
bottleneck in the production of olive oil is thought to be the
malaxation stage. Pectinolytic enzymes boost the phenolic
compounds present by preventing the phenolics from
complexing with the pectic polysaccharides, according to
many studies. However, using A. giganteus extract in the oil
extraction did not result in noticeably higher polyphenol
content. The findings were consistent with those that Peres
et al. have already published. Carotenoids and chlorophylls
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are other significant pigments that affect the look of olive
oil. The composition of the pigments in the olive oil was
unaffected by the pectinolytic enzyme treatment in trials. A.
Giganteus pectinases enhanced the rheological
characteristics and oil output of olive oil without changing
the chemical makeup of the oil. Consequently, the pectinase
extract from A. giganteus NRRL 10 is appropriate for use in
the production of olive oil.
3.3 Applications In Environmental Sector
3.3.1 Wastewater treatment
In the past, numerous processes have been used to treat
wastewater from citrus processing plants that contained
pectic compounds. This has a number of drawbacks,
including increased treatment costs, prolonged treatment
periods, and chemical-related environmental damage.
Activated sludge treatment poorly breaks down pectinaceous
components found in fruit processing industry wastewater by
microorganisms. By utilising alkalophilic bacteria58, attempted
to establish a new method. An alkalophilic Bacillus sp. GIR
621 was discovered from Thai soil, which in alkaline
environment at pH 100 developed an extracellular endo
pectate lyase58. The pretreatment of the wastewater water
with the strain GIR 621-7 succeeded well to get rid of pectic
materials. The endo-PL from GIR 621-7 was believed to
possess a relative molecular mass of 33 000, a pI of 8/8, an
optimal pH of 9, an ideal temperature of 55–60°C, and a
desired Ca2+ concentration of 0–4 mM22.
3.3.2 Bioenergy production
Environmental biotechnological processes’ typical
treatment goal is the removal of contaminating substances to
produce a liquid, gaseous, or solid residue that can be reused
in a natural environment without harming it.
Energy is a fundamental prerequisite for the growth of
practically every part of a culture in the world. However,
using non-renewable energy can result in a number of issues.
Fossil fuel is non- renewable and its overuse will result in a
significant energy problem, which is currently a major issue
for the entire globe. Using conventional fossil fuels can also
produce pollutants like more carbon dioxide and other
greenhouse gases, which contribute to global warming. As a
result, bioenergy, a potent renewable fuel that can replace
fossil fuels, has grown over the past few decades, particularly
in North America and Europe, with the goals of addressing
the rise in global population, and reducing global warming66.
Endopolygalacturonases (EPGs), acetyl – and
methylesterases, – arabinofuranosidase, and -galactosidases
are the pectinases that are most frequently utilised to
hydrolyze pectin. Plants and fungi are the microorganisms that
have been examined the most for bioenergy generation, and
only fungi EPGs have had their 3D structures characterised61.
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V Vaishnavi, G Divyashri, Thomson Tanya, S Sejal and T P Krishnamurthy
3.4 Applications In Pharmaceutical Sector
3.4.1 Preparation of dietary fibre - functional foods
The gastrointestinal system is unfamiliar with foods and
bacteria, although hypersensitivity reactions to dietary
antigens are rare. There is growing evidence that fermentable
prebiotic fibres, including those found in gut-associated
lymphoid tissues, influence the immune system in many ways
as prebiotics. These fibres may influence immunological
alterations via the colon bacteria51. Pectin, which wouldn’t be
broken down in the gastrointestinal system, has found utility
as a fibre by raising intestinal viscosity, which in turn, reduces
cholesterol absorption. Recently, the PME enzyme changed
pectin to produce better gel-forming agents, which are
becoming more and more popular as functional foods.. By
increasing the excretion of fecal bile acids and neutral sterols,
high-viscosity pectin is suggested to reduce cholesterol levels.
Additionally, it might prevent micelles from forming and/or slow
down the rate at which micelles containing bile acids and
cholesterol diffuse through the bolus, which would reduce the
amount of cholesterol and bile acids that are taken in. These
not only shield the intestine against inflammatory illnesses but
also regulate the gut hormones that regulate hunger and insulin
release60. Acetate also appears to be the main short-chain fatty
acid that leaves the liver and enters the bloodstream. As a
result, pectins modified by pectinases are used as functional
additives in a variety of food products for probiotic purposes
3.4.2 Formulation for lowering blood glucose and
cholesterol
Pectic compounds produced by pectinase treatment of
vegetable and fruit peels are being considered as a useful
component in pharmaceutical-based goods because to their
high fibre content. According to reports and claims made by
scientists, pectin may play a role in the treatment and
prevention of serious illnesses like diabetes and obesity;
however, this is fully dependent on viscosity, molecular mass,
and esterification level. More soluble fibres are thought to
increase intestinal viscosity, reducing the amount of bile acids
that are reabsorbed. As a result, the production of bile acids
from cholesterol increases, aiding in the blood cholesterol’s
poor circulation. By using the pectins that have been digested
from the peels of apples, oranges, soybeans, and other plants,
in vivo and in vitro models have been used to investigate
similar herbal compositions5.
4.0 Current Limitations And
Future Prospective
Due to the fact that enzymes have a substantial impact on
nearly every industrial sector (such as food, feed, and
414 | Vol 71(3) | March 2023 | http://www.informaticsjournals.com/index.php/jmmf
medicines), the market is rising quickly to keep up with
consumer demand. Enzyme progress in industrial fields is
hampered, nonetheless, by their stability and high cost. The
enzyme’s resistance to extreme temperatures, unfavorable pH
conditions, and organic solvents is crucial for
commercialization. The employment of enzymes in commercial
operations is constrained by their weak resilience to harsh
industrial environments. To maximize pectinase synthesis, a
variety of methods can be used; however, the cost of general
applicability is higher due to the unstable nature of enzymes.
Because it is challenging and expensive to maintain a
consistent temperature throughout a large-scale bioreactor,
thermophilic enzymes are receiving more interest in research.
One of the most crucial variables is the cost-effectiveness
of generating pectinase from certain microorganisms and
applying environmental parameters. To increase the number
and quality of finished products, pectin hydrolyzing enzymes
are applicable to many industrial operations Examining the
manufacturing process and physicochemical characteristics of
new enzymes is crucial in this strategy. Research should be
done on the immobilization of pectinase enzyme for reuse in
order to further lower the overall cost. Because the alterations
are totally under control, genetic engineering is a much more
effective solution. More investigation is needed to identify
strains that produce pectinase alongside other enzymes, and
each application calls for a precise combination. The focus of
future pectinolytic enzyme research should be on
understanding the molecular mechanisms that regulate enzyme
release and the mechanisms by which various pectinolytic
enzymes function on various pectic substrates
In this manner, well-designed research can offer crucial
tools for controlling microorganisms to create large amounts
of effective and affordable enzymes. All operations in the future
seem to be capable of being completed by pectinases.
Industrial uses of pectinases include the production of textiles,
the processing of fruits, the extraction of oils, and the
fermentation of coffee and tea. The extensive research
demonstrates that pectinases have received priority for the
significant creation or enhancement of enzymes for industrial
purposes. Protein engineering research should be prioritised
in order to create enzymes that are dependable and flexible, as
well as to improve manufacturing processes utilising novel
strains, if this method for exploiting microbial pectinase is to be
effective.
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