A Review on Production of Polygalacturonase Using Various

_____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected];

Asian Journal of Biotechnology and Bioresource Technology 5(3): 1-12, 2019; Article no.AJB2T.52023 ISSN: 2457-0125

A Review on Production of Polygalacturonase Using Various Organisms and Its Applications

A. Radha1, R. Sneha1, R. Kiruthiga1, P. Priyadharshini1 and N. Prabhu1*

1Department of Biotechnology, Vivekanandha College of Engineering for Women, Elayampalayam,

Tiruchengode - 637 205, Tamil Nadu, India.

Authors’ contributions

This work was carried out in collaboration among all authors. Authors AR and RS designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript.

Authors RK and PP managed the analyses of the study. Author NP managed the literature searches and statistical analysis. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/AJB2T/2019/v5i330063

Editor(s): (1) Dr. Md. Shaheed Reza, Department of Fisheries Technology, Agricultural University, Bangladesh.

Reviewers: (1) Panan Kanchanaphum, Rangsit University, Thailand.

(2) Mariana-Atena Poiana, Banat’s University of Agricultural Sciences and Veterinary Medicine, Romania. Complete Peer review History: https://sdiarticle4.com/review-history/52023

Received 03 August 2019 Accepted 14 October 2019

Published 30 October 2019

ABSTRACT

Polygalacturonase is a pectinolytic enzyme that catalyses the hydrolytic cleavage of the polygalacturonic linkage chain. An enzyme is a polygalacturonase is expressed in fruits. The polygalacturonase produced from various organisms isolated from various fruits. The solid-state fermentation was used in the production of polygalacturonase. The production of PG was found at various incubation period and pH and temperature are using fruits as best nitrogen and carbon sources. Although they have other parts of the genome they are active in the fruit. Peak expression requires a full range of promoter and saturation lines. Based on the physicochemical properties of the purified enzymes, this enzyme possesses great potential for industrial and biotechnological application such as oil extraction, fruit clarifications.

Keywords: Polygalacturonase; microorganism; fermentation; optimum culture conditions.

Review Article

Radha et al.; AJB2T, 5(3): 1-12, 2019; Article no.AJB2T.52023

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1. INTRODUCTION

Polygalacturonases (PGs) (EC.3.2.15) are an important member of pectinase with catalysing the hydrolysis of homogalacturonan or polygalacturonic acid into galacturonic acid monomers by breaking of α- 1, 4-linkages. polygalacturoanse (PGs) is the major enzyme responsible for the depolymerisation of cell walls and softening of fruit tissues [1]. Polygalacturonase also is known as pectin depolymerase, PG, pectolase, pectin hydrolase and poly-alpha-1, 4 galacturonide glycanohydrolase, is an enzyme that hydrolyzes the alpha-1, 4 glycosidic bonds between galacturonic acid residues. The majority of the enzymes are derived from fungi and bacteria. PGs are widely distributed among fungi, bacteria, yeast, plants and some plant-parasitic nematodes. In the food industry, enzymes are used in extraction to increase the yield of fruit and vegetable juices, to control the clarity of juices [2,3]. Polygalacturonase appeared to be the most frequently encountered pectin enzymes. They are formed in the majority of plant tissues, particularly in repining fruits. Also saprophytic microorganisms and plant pathogenic produced polygalacturonase (Parvateesam and Verma, (1992), Ajayi et al. (2003). Polygalacturonase exists in two forms; endopolygalacturonase act randomly on the alpha-1, 4-polygalacturonic backbone chain.exopolygalacturonases act on the non-reducing end of the alpha-1, 4-polygalacturonic backbone chain. polygalavturonase use in many industries. It is also found in plant tissues particularly in ripening fruits and is also produced by a microorganism such as fungi and bacteria mixtures of polygalacturonase [4]. In agro-industries, they are used for the removal of settled or suspended solids that presents a cloudy appearance of juices especially the acidic ones [5].

The enzymes are also employed in coffee fermentation and extraction of oils or retting of fibres [6]. Furthermore, PGs are used in food industries for productions of jams, jellies, frozen foods, and more recently in low-calorie foods as fat and/or sugar replacement(s) [7,8]. Generally, the applications of polygalacturonase to food and fruit industries have solved several problems that have arisen during extraction, filtration, and clarification of fruit juice and also in biotechnological experiments. Polygalacturonase produced by two methods: Solid-state and submerged fermentation and extracted by centrifugation and filtration [9].

Polygalacturonase (PGs) are produced by various organisms, such as plants, bacteria and fungi. The pg activity was a large difference in fruit for example; Avocado fruit has high levels of PG activity that are temporally correlated to solubilisation and depolymerisation of polyuronides during fruit ripening. Green tomato fruit has low levels of pg activity is present in exo-acting. PG isoform that is expressed constitutively throughout development and ripening. pg activity increases exponentially during fruit ripening as a result of the de novo synthesis of endo-acting PG isoforms. The majority of research has focused on PGs in ripening fruit, abscission zones or pollen. PGs have first identified over 35 years ago and have been suggested to be involved in the disassembly of pectin that accompanies many stages of plant development, particularly those that require cell separation [10]. For example, PG activity is associated with organ abscission, pod and anther dehiscence and pollen grain maturation and pollen tube growth. In this ripening system, the initiation and early stages of fruit softening were accompanied by a decrease in the molecular size of hemicellulosic polysaccharides, most notably of a tightly bound fraction of xyloglucan [11]. Also, it is now clear that PGs are encoded by relatively large gene families in plants and expressed in a wide range of developmental contexts than previously appreciated [12]. Structural variation has been identified among differing PGs depending on organisable [13]. Origins and catalytic functions. For example, endo –Polygalacturonase produced from Erwinia carotovora demonstrate functional similarity to pectate lyases in that backbone, but some of the monomers are methyl esterified on the sixth carbon. PG acts on pectate, not pectin [14]. The PG activity was a large difference because of various microbes present in fruits (such as tabulation1). Then many types of research are proving that to absent the PG activity during fruits is repining and softening. PG is not only used the softening process. Additionally, these enzyme activity cultivars the different type of in same fruit. Polygalacturonase has a share of 25% in the global sales of food enzymes and has numerous biotechnological application such as in fruit juice extraction and its clarification, bleaching of paper, scouring of cotton, wastewater treatment, vegetable oil extraction, tea and coffee fermentation in poultry feed as additive and in

alcoholic beverages, pectinase is enzyme that degrades pectin. They are mainly divided into esterase or polymerase based on their actions. One of the depolymerise enzyme polygalacturonase. PG catalyzes the hydrolytic cleavage of polygalacturonic acid linkage with the introduction of water across the oxygen bridge in the smooth region of pectin. Among the family of pectinolytic enzymes, PG is the most extensively studied. They are found in plant tissue particularly in ripening fruits and are also produced by microorganisms such as fungi and bacteria. Mixtures of polygalacturonase, pectinesterase and pectate are available as commercial enzyme preparations and are used in processing food industries.

1.1 Structure Many synonyms beta-sheets of this enzyme are a helical form, called a beta-helix. This highly stable structure, to the many hydrogen bonds and fibres between the disulfide bonds, is a common characteristic of the enzymes involved in the degradation of pectin [15]. The betaof the interior is hydrophobic [16]. Xcrystallography has been used to determine the three dimensional (3D) structure of PGs in organisms such as Fusarium moniformeactive site Fusarium moniliforme PG comprises 6 charged amino acid residues: H188, R267

Fig. 1. Image of polygalacturonase magenta

Radha et al.; AJB2T, 5(3): 1-12, 2019; Article no.

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alcoholic beverages, pectinase is enzyme that degrades pectin. They are mainly divided into esterase or polymerase based on their actions.

depolymerise enzyme PG catalyzes the hydrolytic

cleavage of polygalacturonic acid linkage with the introduction of water across the oxygen bridge in the smooth region of pectin. Among the family of pectinolytic enzymes, PG is the most

tensively studied. They are found in plant tissue particularly in ripening fruits and are also produced by microorganisms such as fungi and bacteria. Mixtures of polygalacturonase, pectinesterase and pectate are available as

and are used

sheets of this enzyme are helix. This highly

stable structure, to the many hydrogen bonds and fibres between the disulfide bonds, is a common characteristic of the enzymes involved

in [15]. The beta-helix of the interior is hydrophobic [16]. X-ray crystallography has been used to determine the three dimensional (3D) structure of PGs in

Fusarium moniforme. The PG comprises 6

mino acid residues: H188, R267 and

K269 are involved in substrate binding, D212 is responsible for proton donation to the glycosidic oxygen and D213 and D191 activate Hnucleophilic attack.

1.2 Mechanism Polygalacturonase enzyme that degrades pectin. PGs hydrolyze O- glycosyl bonds in the pectin polygalacturonase network [17]. The rate of hydrolysis depends on the length of the polysaccharide chain. Low rates of hydrophobicity are associated with very shorchains (e.g. decalcuturonic acid) and verychain [18].

1.3 Exo-vs. Endo-polygalacturonase EXO-and ENDO –PGs use different hydrolytic systems. Endo –PGs hydrolyze at random with a polygalacturonase network. This pattern results in oligogalacturonase [19]. The (Fig. 2) EXOhydrolyze at the non-reducing end of the polymer to form monosaccharide galacturonic acid. Occasionally, organisms use both methods. In addition to the different types of processes, BG is also available. Polymorphism us fungi PGs can effectively degrade a wide range of plant tissues.Bg type, substrate specification and other factors at optimal PH may contribute to phytopathogenic organisms such as fungi [20].

polygalacturonase from Fusarium moniliforme (1HG8) showing active site in magenta. Molecular formula: C43H67N15O12S2

; Article no.AJB2T.52023

K269 are involved in substrate binding, D212 is responsible for proton donation to the glycosidic oxygen and D213 and D191 activate H2O for a

Polygalacturonase enzyme that degrades pectin. glycosyl bonds in the pectin

polygalacturonase network [17]. The rate of hydrolysis depends on the length of the polysaccharide chain. Low rates of hydrophobicity are associated with very short chains (e.g. decalcuturonic acid) and very-long-

polygalacturonase

PGs use different hydrolytic PGs hydrolyze at random with a

polygalacturonase network. This pattern results oligogalacturonase [19]. The (Fig. 2) EXO-PGs

reducing end of the polymer to form monosaccharide galacturonic acid. Occasionally, organisms use both methods. In addition to the different types of processes, BG is

rphism us fungi PGs can effectively degrade a wide range of plant tissues. Bg type, substrate specification and other factors at optimal PH may contribute to phytopathogenic

(1HG8) showing active site in


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Table 1. Polygalacturonase activity from various organisms

Organism Substrate Kinetic parameter Mra

(kda) Optimum pH

Optimum temperature

Stability temperature & pH References

Rhizomucor pusillus

Orange peel Km-0.22 mg/Ml, Vmax-4.34U/Ml,

32 5 55 n.d [41]

Aspergillus niger MTCC 478

Orange juice Km-2.3 mg/Ml, Vmax- n.d

~124 4.0 50 3.0-11.0 & 10-40ºC

[40]

Aspergillus carbonarius

Apple Km-n.d, Vmax- n.d 42 4.3 10 min 50ºc n.d [43,44]

Aspergillus fumigatus MTCC2584

Crotalaria juncea plant

Km-2.4 mg/Ml, Vmax- n.d

~43 10.0 30 7.0-11.0& 10-30

0c for 2h

[36,45]

Bacillus subtilis Soil Km-n.d, Vmax-n.d n.d 4.0 60 9.0 & 600C [46]

Aspergillus flavus Orange peel Km-0.705 mg/Ml, Vmax-1.0508µmol/min

554 4.5 35 n.d [42]

Aspergillus AN07 Papaya, orange peel

Km-2.6 mg/Ml, Vmax-181.8µmol/min

64.5 55 5.0 3.0-5.0 & 550C for 1 h [33]

Aspergillus niger Banana peel Km-n.d, Vmax-n.d 32 5.0 40 3-7 & 20ºC

[31,32]


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Fig. 2. The mechanism of polygalacturonase: Hydrolysis of polygalacturonan

2. PRODUCTION OF POLY-

GALCTURONASE FROM BACTERIAL STRAIN BY TOMATO

The polygalacturonase is expressed in tomato fruit. The polygalacturonase is the best enzyme of ripening the tomatoes. The role of this enzyme is the degradation of the cell wall in tomatoes. By the action of this enzyme, the tomato is converted into a fruit. We have many advantages of this invention. The polygalacturonase is important for the retention of tomatoes. Although their polygalacturonase linkages are present in many parts of the crop, only the fruit is unique [21]. This enzyme is the most uses for industry [22]. The microorganism is a most useful source of this enzyme [23] Tomatoes fruits are best nutrients in our body and tomato fruits component is used in our body health and growth and development and easily attack the microbes in tomato because of the PH and moisture content involved in large variance of the chemical component [24]. Saxena and R.gupla are said that, isolate the total bacterial strains that produce Polygalacturonase enzyme from tomatoes fruits [25,26].The total bacterial strain such as Aeromonas sp., Klebsiella sp., Citrobacter sp., Enterobacter sp., Staphylococcus sp., Lactobacillus sp. and Micrococcus sp. The Saxena and R. Gupla, et al. are used (method) materials such as collection the sample. 1st fresh tomatoes fruits are purchased

from the market. There are 200 fruits are need, because the first 100 fruits are temperature condition and another 100 fruits are refrigerator condition. The tomatoes are sorted, then washed and in the laboratory. The first method is there are 100 fruits are stored AT temperature condition. The second method, is 100 fruits are stored CT at the refrigerator. Both 200 fruits method contained the 15 days.After the process is to isolate the bacterial strain (Aeromonas sp., Klebsiella sp., Citrobacter sp., Enterobacter sp., Staphylococcus sp., Lactobacillus sp. and Micrococcus sp). In tomatoes and the population of bacterial washing, the method is to pour plate. The different sample of tomato fruit such as tomato juice and the extract was diluted in distilled water. Because every method was properly handled and sterile water is not there any other contamination. The using the agar was nutrient agar (NA) is incubating at 37ºC 24h. After the bacterial strain was identification in basic microbiology and using the scheme of bacteriology of Bergey's manual [27]. To use the medium is mostly different temperature condition and to use the pH value is different concentration. After determining the polygalacturonase activity.The polygalacturonase incubated at 35ºC for 3h [28] (Tabulation 1). The Saxena and R. Gupla were given the conclusion that the population of bacteria. The total bacterial are grow on tomato as room temperature and refrigerator method was the production of polygalacturonase. The different type of production of polygalacturonase Such as


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bacteria, fungi, yeasts. Maybe this enzyme is isolated from other sources. Like bread (yeast), vegetables (fungi), potato (lactobacillus), soil (algae) are described.

The ripening process reoccurs in conjugation and increases the production of ethylene and CO2 in tomato fruit [29,30]. Production of Polygalacturonase from tomatoes is high. In this property is the helpful producing role of the enzyme in sources. Like food, drink and industry. 3. PRODUCTION OF POLY-

GALCTURONASE FROM Aspergillus niger BY BANANA

Sumi barman and Nandan sit & laxmikant s.badwaik & sankar c.deka is to prove that production of PG from banana. The banana is the second-largest fruits in India and it is used to source of food. It is highly active polalioun, niacin and riboflavin. PG is produced from the production of banana. The bananas juices are mostly highly qualified production of PG. The polygalacturonase is used in fruit industrial and food industry (patil and dayanand 2006) [31]. The banana was production method is the different way such as juice, peel and powder. The powder was production enzyme such as PG. The agricultural and industrial wasted such as banana peel was purified and production the PG enzyme (Martin, et al. 2004) [32]. The powder is dried because the moisture content is evaporated by dried heating of the oven method. To isolate the organism are based on its morphological, colonial and microscopic examination. The solid-state fermentation was found to be more suitable for PG production by Aspergillus Niger. Various type of waste peel of fruit and vegetables are used for the production of polygalacturonase (taskin (Elton 2008; Bari et al. 2010; kuneear et al. 2012) [33]. Mostly different banana are produce polygalacturonase (Ramli, et al. 2009, dhabacker and chandak 2010) [34]. The banana juice is extracted for hot water. The production of polygalacturonase is used for the banana fruit and the stability storage was increased for example of ripe banana is mainly used for juice extraction. Sumi barman and nandan sit & laxmikant s.badwaik & sankar c.deka use the method is a collection of banana after wash and to cut in piece. (1% of banana and 2% of distilled water) 1:2 ratios are mixed. This juice was mainly used

productional of PG. It is a nutritive and medicinal value. Mostly grows well in a temperature range of 15ºC – 35ºC with relative humidity 75-85% and pH between 6.5-7.5. The Aspergillus niger culture was purchased (banana) in institude. The agar medium using cultivated the culture at temperature of 30ºC and 7 days are incubated. After the potato dextrose agars are the maintained the culture and every 30 days .The tab water was used to wash the banana peel. After dried the peel at 45ºC for 24 hour. The dried peel is grinted. Finally to the bowder was there. The bowder was used production of polygalacturonase. 0.5% Polygalacturonase acid was incubating by Polygalacturonase activity (svgma Aldrich, USA). The citrate buffer was used in 0.2 M (pH 5.8) and these enzymes are extracting at 37ºc for 60 min. The nelson sonogyi method (smogy 1952) [35]. It was determined the this enzyme activity.it also export polygalacturonase activity increases initially with the increase in substrate concentration, time of incubation and temperature of incubation and then decreases with further increase in the processing parameters within the experimental range. Similar results were obtained by Mukesh Kumar, et al. (2012) (Tabulation 1). The Francis sopuruchukwu lre and Emmanuel Garba vinking are the productions of polygalacturonase from the same organism in solid-state fermentation and submerged fermentation using a banana peel. The production of polygalacturonase in submerged fermentation was carried out in a 0.6% (NH4)2SO4, MgSO4.7H2O & 0.6% KH2PO4 are liquid chemicals are prepared of basal medium and the banana peel are dried to taken 2% was added to the basal medium to sterilized the medium was 121ºC for 16minutes. After inoculated and this is used to isolate of pectolytic .then the flask were properly handled and 5 days 30ºC for incubated .many temperature condition method was measured the crude enzyme. Finally, the enzyme was assayed the submerged fermentation method. The production of polygalacturonase in solid state fermentation was carried out in the banana peel was weighted for 15 g and 0.6%(NH4)2 SO4, 0.6% K2HPO4 and MgSO4.7H2O 0.01% of the mineral salt solution was prepared for The Francis sopuruchukwu lre and Emmanuel Garba vinking. The solid-state of the medium was sterilized at 121ºC for 40 min [18]. Inoculated


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with the test isolate into the flask and 30ºC for incubated for 4 days. Then the Francis sopuruchukwu lre and Emmanuel Garba vinking were added the 50 millilitres of distilled water in the flask at 4 days and then filtered. Finally conducting PG assay was filtrate method. Effect of carbon sources on enzyme production carried out in the set of to add the 1% of glucose, starch, fructose, mannitol & cassava peel in the fermentation medium in best carbon source for fermentation. Finally, the fermentation process method was measured the PG enzyme activity [36]. Effect of nitrogen sources on enzyme production carried out, of fermentation medium because this medium is best nitrogen source for fermentation .the medium is obtained by 0.6% NH4cl, NaNO3, peptone, yeast extract KNO3 and urea were separated added to the medium. 4. POLYGALACTURONASE

PRODUCTION FROM Aspergillus flavus BY ORANGE PEEL

The author of Daughari JH and onyebarachi GC are said that the production of PG from fungal. This fungal one of the natural sources. This type of fungi is produced higher PG activity on a selective medium containing pectin. Mostly this type of fungi on grown on orange peel. The orange peel helpful because decrease the cholesterol in our body and the blood pressure is controlled and in this peel helpful of weightless because vitamin C is present it. Orange peels can be used effectively in the production of PG under solid state fermentation system using Aspergillus flavus. The ability of the Aspergillus flavus strain to degrade pectin demonstrated by the production of PG.

The author of Daughari JH and onyebarachi GC are carried out in a 2 g soil sample were collected and transferred the McCartney bottle was taken because the soil sample was transferred the bottle. After the sample was diluted with distilled water and sample were inoculated using the spread plate method using PDA (Potato Dextrose Agar) medium and 30ºc for 7 days incubate the culture plate. After to isolation of fungi [37]. The isolate the fungi were inoculated onto the Petri dish of MCDA media was used. later, incubated at 30ºC for 24-96 h and the plate was drawn with 50 Mm iodine [38]. The colony is present in Petri dish. Finally, the fungal strain was the production of polygalacturonase [39].

The conclusion of this author was to isolate the three fungal from natural sources. To isolate the Aspergillus flavus with genetic profile size of 554 bases were confirmed the microscopic and molecular characterization. The maximal production of PG is orange peel because using the substrate was carbon followed by plantain peel and yam peel. The enzyme production is decreasing because the glucose was used as carbon sources. Organic and inorganic nutrition is mainly used for PG production. The production of PG by fungal was obtained in the presence of ammonium sulphate. To compare the organic is higher production of PG in inorganic nitrogen source. The maximal production recorded on day 4 during the effect of incubation period also showed that production of PG is increasing. The PG activity temperature condition is 35ºC and pH 4.5 and SDS-Page with a molecular weight is 66KDA which depend on aspergillus Flavus. Ascorbic acid was the strongest inhibition of enzyme because of the inhibitory effect of acid on purified polygalacturonase [40]. Respectively, Km and Vmax were also found to be 0.705 mg/Ml and 1.0508 µmol/min. Based on the PG properties of the purified enzyme, this enzyme possesses great potential for industrial and biotechnological applications such as fruit juice clarifications, oil extraction. Orange peels used in this process can be considered as a cheap substrate for effective and resourceful production of PG using Aspergillus flavus because not only do they contain a significant amount of pectin, they are readily obtained with little or no cost. This will minimize the production cost of PG and also the amount of pollution of the environment by this waste (Tabulation 1). 4.1 Effect of Carbon Sources on PG

Production Many wastes peel and chemicals were used in the production of the medium. The peel such as orange, banana, yam and plantain and chemicals such as glucose, sucrose and pectin which concentration of carbon source have checked the production of the medium. The PG activities have assayed the cultures.

4.2 Effect of Nitrogen Sources on PG Production

The Nitrogen source substances are urea, casein, yeast extract, peptone and (NH2)4SO4 are basal medium. To check and test their effect on PG activity.


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5. POLYGALACTURONASE PRODUCTION FROM Rhizomucor pusillus BY ORANGE PEEL

The Rhizomucor pusillus is a fungus and mostly this fungi was growing on soil, fruit and vegetables. This organism is can grow at temperature condition is ~54ºC. Mohd. Asif siddiqcci, veena pande and mahammad Arif et al. The production of PG carried out of the different type of fungi are selected and produced in PG were isolated from different soil, vegetables wastage and fruits by pectin agar medium are used by the composition and mineral composition. To make the 1 litter distilled water and to adjust the pH value is 6.5-6.0 by using NaOH.this medium is sterilized with 121ºC for 15 min. The medium was inoculated plates and 50ºC for 5-7 days incubate it. The colonies are indicating of a pectinase. Finally the fungal were isolated characters following Dubey &maheswari. The author was the production of enzyme by using solid-state cultivation method was used the solid-state cultivation is an orange peel was carried out in 50 ml. The 15 g of the basal medium is inoculated with a 5-day slant. The pH was adjusted to 5.5-6.0 and temperature 50ºC for 4-5 days. To extract the fermented media with 30 ml of distilled water. To shaken the flask and filtered and 100 ml of citrate buffer was add the extract. To centrifuge the extract at 10000 rpm for 15 min at 4ºC [41].

Different fungi strains are isolated from different soil, fruits and vegetable sample and different spices are isolated. The author is isolating the 40 fungal strains in various soil, fruit, and vegetable samples. Different species of Mucor, Aspergillus penicillium, Rhizopus and Trichoderma were isolated. High pectnase producing strains were further screened semiquantitatively by plate assay method. The author is isolating the maximum activity of PG in Rhizomucor pusillus. The isolate was further identified as Rhizomucor pusillus finally isolate are the high activity of PG and identification as Rhizomucor pusillus. To analyze the temperature and pH activity from Rhizomucor Pusillus at 55ºC and 5.0 [42].

The tabulation 1 are explained the polygalcturonase activity such indicated of Rhizomucor pusillus are grow on optimum temperature & pH, substrate, stability

temperature and pH. The maximum high activity of production. The Aspergillus niger MTCC478 (fungal strain) was used for the production of polygalacturonase both under submerged and solid-state fermentation condition. Further its production was optimized under solid-state fermentation condition with media comprising of wheat bran and tea extract.

6. POLYGALACTURONASE PRODUCTION FROM Aspergillus niger MTCC 478 BY SOLID STATE FERMENTATION

Aspergillus niger is one of the most common fungi in the environment saprophytic fungi and the primary causative agent of disease in immune-compromised individuals. It is also the source of several bioactive compounds.

The authors (Guatam Anand, Sangeeta Yadhav et al.) are determined by production of polygalacturonase from fungal strain Aspergillus niger MTCC 478 was grown in 10 250 mL Erlenmeyer flasks containing M3 media. The media was inoculated with 1 mL of spore suspension in each flasks were kept at 26ºC in a biological demand incubator (BOD). Enzymes produced from the fungi Aspergillus, Rhizopus and Penicillium are generally regarded as safe (GRAS) and produce extra cellular enzymes which can be easily recovered. High clarity and low viscosity have high commercial values of substrate are used an industry such as vegetable and fruit juice industry, juices. Fruit generally contains pectin and other polysaccharides leading to fouling and colloid and substantially influents the commercial value of juices. Microbial polygalacturonase have been play role in viscosity and reduction and clarification of juice. In general microbial polygalacturonases are important group of enzymes with potential application in various industries like wine industry, paper industry, textile industry and food industry. Application of purified PGs in clarification of fruit juice was studied on orange juice produced from the local market [43,44]. Maximum production was obtained with medium M3 followed by medium M2. Tea extract was found to increase the enzyme. The optimum pH for the production of polygalacturonase was found to be 4.0 and the enzyme was stable for a wide pH range, 3.0-11.0. The optimum Temperature for the production of polygalcturonase activity was found to be 50ºC and stable for a PG range 10 & 40ºC for 30 min. (Tabulation 1).


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7. INDUSTRIAL APPLICATION OF POLYGALACTURONSE AND PURPOSE

Polygalacturonase have many applications such as clad stabilizations, extraction of juice and oil, maceration, liquefaction, gelatin, wood preservation, retting of fiber crops, Degumming of fiber crops, coffee and tea fermentation. In industries clad stabilization used to precipitate hydrocolloid matter present in fruit juices. Fruit juice clarification is used to Degradation of cloud forming pectic substances. Hence, the juice can easily filter and processed. Extraction of juice and oil is overcome the difficulty in pressing fruit pulp to yield juice and oil. Maceration is used for breakdown the vegetable and fruit tissues to yield pulpy products uses as base material for juice, nectar as in the case of baby food, yogurt, and pudding. Liquefaction in idustries breakdown fermentable plant a carbohydrate to simple sugars.Gelatin use in gelling low-sugar fruit products. Wood preservation is used for prevent the wood from infection by increasing the permeability of preservative. Retting of fiber crops to release fibres from the crops by fermenting with microorganism, which degrade pectin. Degumming of fiber crops used to remove the remie gum of remie fiber. Waste water treatment is degrading pectic substance in waste water from citrus processing industries. Coffee and tea fermentation is used to increase the tea fermentation and foam forming property of tea. Enzymes are added in the high yield because preesing of juice and to remove suspended matter to give sparkling clear juice. The high activity of PG enzymes is added to fruit juice to stabilize the cloud of citrus juice. These enzymes are majorly using in unicellular products examples of transformation of organized tissue into a suspension of intact cells, resulting in products used as base material for pulpy juice, yogurt, and baby food. The waste water from the citrus processing industry contains PG materials barley decomposed by microbes because the activated sludge treatment.

8. AGRICULTURAL APPLICATION OF POLYGALACTURONASE

8.1 Fruit Ripening

Delayed ripening of the first GM food genetically modified tomato (also known as flower saver) available in stores in achieved by preventing the degradation of polygalacturonase pectin, which stabilizes tomatoes. An antisense PG gene was

introduced which prevents polygalcturonase from inhibiting tomato ripening and softening. Pectin depolymerisation in the later stages of fruit ripening. Especially when the fruit becomes excessive [45]. Although tomatoes are a prime example of high BG activity, the enzyme is very active in butter and peach ripening. The PG enzymes in peach, two exo-PGs and one endo –PG, are active when the fruit.

8.2 Abscission

It is often whether PGs play a role in helping some plants move away, and if so, whether they are exo – endo –acting. Contradictory research has been punlished on wheather PGs is involved in the elimation to measure exo-PG activity [46]. Additional complications is the difference in PG enzymatic activity between the fruit and eat cell dividing regions. In peach, PG activity was detected only in fruit beaibg regions.

8.3 Pollen

Exo-PGs play a role in activating pollen tube length because pectin rearrangement is necessary for the development of pollen tubes. This PG activity is found in maize-like grasses and trees; especially the eastern cotton tree.Exo-PGs involved in pollen tube development requires Ca2+ for maximum enzyme activity and may be inhibited by high concentrations of NaCl, citrate and EDTA.

8.4 Other

Agricultural insects such as Lycus Hesperus can damage cotton and other crops because they secrete PGs in saliva. This digests plant tissue. They use exo-and end o-PGs.

9. CONCLUSION Polygalacturonase produced from fungal spices is vary with temperature and PH because depend on carbon and nitrogen source. Production of polygalacturonase can be an efficient and economic enzymatic option for food industries. Based on the physicochemical properties of an purified enzyme, this enzyme possesses great potential for industrial and biotechnological applications such as fruit juice clarification, oil extraction. The various fruits used in this process can be considered as a cheap substrate not only does they contain a significant amount of pectin; they are readily obtained with or no cost. This will minimize the production cost of polygalacturonase and also the amount of pollution of the environment by this waste.


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COMPETING INTERESTS Authors have declared that no competing interests exist.

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