Nishank Prakash et al.

, IJSID, 2012, 2 (2), 266-273

ISSN:2249-5347

IJSID

International Journal of Science Innovations and Discoveries

Research Article

An International peer Review Journal for Science

Available online through www.ijsidonline.info

PRODUCTION OF ECONOMICALLY IMPORTANT PRODUCTS BY THE USE OF PULLULANASE ENZYME

Nishank Prakash, Sweta Gupta, Moinuddin Ansari, Zaved Ahmed Khan, Vuppu Suneetha*
School of Bio-Sciences and Technology, VIT University, Vellore, Tamil Nadu, India

Received: 06.02.2012 Accepted: 30.04.2012

*Corresponding Author

ABSTRACT industries for the production of various economical products. Pullulanase is also known as debranching enzyme and is one of the most important enzymes used for starch modifications. Pullulanase hydrolyse -1, 6-glycosidic linkages of polysaccharides. It is a very powerful enzyme for the degradation of starch to glucose or maltose. This property of the pullulanase enzyme is widely used for the large scale production of glucose and The study on pullulanase is a recent research for implications in various

Address: Name: Dr. V.Suneetha Place: Vellore, TamilNadu, India E-mail: vsuneetha@vit.ac.in

maltose syrup and also in the baking industry. Also pullulanase is quite inexpensive and easily available making it useful for the economical production of cyclodextrins, bioethanol and beer in the industries. Resistant starch and substitute of natural gums are possibilities. pharmaceutical and neutraceutical applications. With the improvement in the stability and

also produced INTRODUCTION by action of pullulanase; these products have various potential activity of pullulanase, it could be used in various industries with new applications and

INTRODUCTION Keywords: Pullulanase enzyme, -1, 6-glycosidic linkage, starch.

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Nishank Prakash et al., IJSID, 2012, 2 (2), 266-273 hydrolyses pullulan, which has the repeating units of maltotriose joined together in head to tail fashion by -1, 6 glycosidic bonds. Since pullulanase acts on the branch points of the polysaccharides is also known as debranching enzyme. Pullulanase microorganisms [1].
[2].

also attacks on -1, 4-glycosidic linkage with other residues.Pullulanase which attack both -1, 6 glycosidic bonds and -1, 4glycosidic bonds in dextrin, amylopectin and malto-oligosaccharide have been isolated from various thermophilic -1, 6-glycocidic linkages but pullulanase type II specifically attack on -1, 4-glycocidic linkage and -1, 6-glycocidic linkage The highly thermostable pullulanase type-II enzyme is the most interesting from industrial point of view which is obtained shows maximum enzymatic activity at pH 6.0 and temperature 55C [9]. The -1, 6-glycosidic bonds are easily broken by the pullulanase enzyme and this property is widely used in the industries for the productions of various important products. APPLICATIONS OF PULLULANASE ENZYME a) In the food processing industry Pullulanase obtained from Bacillus stearothermophilus is highly thermostable and acidophilic. This kind of pullulanase

Pullulanase (pullulan - glucanohydrolase; EC 3.2.1.41) is an extracellular carbohydrase enzyme .Pullulanase INTRODUCTION

spinach [4] and millet [5]. The enzyme is produced by several bacteria also like Klebsiella pneumoniae and Bacillus species [6][7]. from hyperthermophilic archaea, for example members of genera Thermotoga maritima and Pyrococcus etc.[8].Pullulanase

Various bacteria and plants act as the source of pullulanase enzyme. The plant sources include endosperm of rice seed [3],

The pullulanase enzyme is of two types, pullulanase type-I and pullulanase type-II. Pullulanase type-I hydrolyses only

is used in sugar syrup industries for the complete hydrolysis of starch which is started by -amylases. The combination of amylase and pullulanase simultaneously increases the efficiency of the starch saccharification reaction which in turn increases the quality of sugar syrups.
Starch granules

Starch slurry Gelatinisation Bacterial -amylase Gelatinised starch Liquefaction Liquefied starch D-glucose Maltose Oligosaccharides
glucoamylase pullulanase

Saccharification

fungal -amylase pullulanase

D-glucose(high content)

Glucose syrup

International Journal of Science Innovations and Discoveries, Volume 2, Issue 2, March-April 2012

Fig 1 general outline for the production of glucose and maltose syrup

Maltose isomaltose Other oligosaccharides

Maltose(high content)

Maltose syrup D-glucose

Isomaltose Other oligosaccharides

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[11],[12].Pullulanase

sweeteners by debranching of corn starch b) In the baking industry

saccharification[15] time in the manufacture of monosodium glutamate, alcohol, vinegar and soy sauce. The general method for the production of glucose syrup and maltose syrup is given in fig 1. Starch and starchmodifying enzymes are used by baking industries in large amounts around the world. One of the

This method is highly advantageous for the generation of higher yields of a desired end product from starch. [10][7] enzyme is also in use as a processing aid in the food industries, like in the production of certain corn
[13]. [8], [14].

Nishank Prakash et al., IJSID, 2012, 2 (2), 266-273

Pullulanase is also use to improve the glucose content and shorten

major problem that is faced by baking industry is the staling effect [16] .The staling effect leads to increase in the firmness of crumb, loss of crispness of the crust, reduction of moisture content of the crumb and flavour of the bread is changed. This anti-staling treatment[18], [13] nowadays the consumers prefer the enzymatic treatment as chemical possess certain health hazards. On the other hand enzymes are produced naturally and are highly accepted by the consumers. Pullulanase plays a major role in the enzymatic c) In the production of cyclodextrins (cds)

greatly reduces the quality of bread and other products [17]. This problem is mainly solved by the chemical treatments. But

members of cyclic oligosaccharides family. CDs mainly compose of 5 or more -D-glucopyranoside units joined together by 1, 4-glycosidic linkages. These are classified into three groups -cyclodextrin: having six sugar ring molecule -cyclodextrin: having seven sugar ring molecule -cyclodextrin: having eight sugar ring molecule

Cyclodextrins are the compounds which are made up of sugar molecules joined together in a ring hence are the

-cyclodextrin

-cyclodextrin

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modifying enzymes. Commonly cyclodextrin glycosyltransferase (CGTase) is used with -amylase. But the -1, 6-glycosidic bond of the amylopectin blocks the action of the CGTase cyclodextrins (CDs) like maltosyl-CDs and d) In the production of bioethanol cholesterol free products in the food industry glucosyl-CDs[21]. increasing the percentage yield of the cyclodextrin production materials, masking odors, and as a solvent for insoluble or poorly soluble drugs. Also these are used for the production of The, CDs and branched CDs are widely used as stabilizers for labile
[19]. [20].

The production of cyclodextrins is quite simple and involves treatment of ordinary starch with a set of starch Also pullulanase is used for the production of branched

Fig 2 .Chemical structure of three cyclodextrins

-cyclodextrin

These bonds are broken by the pullulanase enzyme hence

Fig 3 Duckweed International Journal of Science Innovations and Discoveries, Volume 2, Issue 2, March-April 2012

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slow-moving fresh water bodies. It is also known as "bayroot". It can grow in all seasons in the areas of warm climate. Also its biomass gets double within two days under the optimal conditions the dry weight depending on strains and species of the duckweed effluents [23]. Because of its tolerance to high nutrient levels and preferred absorption of ammonium, duckweed has also been
[25]. [22].

Duckweeds, or water lentils, are small aquatic plants which are found floating on or just beneath the surface of still or Duckweed is also used for the treatment of domestic

Nishank Prakash et al., IJSID, 2012, 2 (2), 266-273

successfully used to recover the nutrients from pig effluents [24]. The total starch content of duckweed can vary from 3-75% of and the concentration of duckweeds play a major role in the accumulation of starch in the duckweed. For example some species like Spirodela polyrrhiza when grown on swine wastewater and grown anaerobically contains a starch content of almost 59%[26][27]. Hence they act as a good source for the production of bioethanol. The duckweed biomass is hydrolysed by for the production of bioethanol from the high-starch duckweed. e) In the brewing industry acts to produce alcohol. Also other factors like availability of various nutrients

the action of pullulanase enzyme and amyloglucoside to produce reducing sugars [28]. These sugars are then converted into ethanol. The overall starch conversion rate by the action of pullulanase is very high; hence it is economical and easy method yeast fermentation process of sugars. Barley contains starch which is first broken down to fermentable sugars on which yeast Beer is an alcoholic beverage which is part of our social life for thousands of years. It is produced form barley by the The beer is produced by mixing hot water and crushed barley malt in a mash. This process is known as mashing.

During mashing the starch is broken down into dextrin and fermentable sugars. After mashing, the mash is filtered to form a

liquid known as sweet wort. Hops are boiled with the sweet wort to produce hopped wort. The yeasts then act on the hopped wort to ultimately produce beer. Pullulanase is added during the mashing step of beer production. Pullulanase enzyme causes the manufacture of beer[31]. This causes improvement in the quality of beer. f) In the production of food gums increasing the percentage yield of the low caloric beer production [30]. Also pullulanase reduces the amount of dextrin during Natural gums are polysaccharides obtained from natural sources. When these natural gums are added even in small

the hydrolysis of the -1, 6 branch points of starch which leads to the maximum fermentability of the wort[29] .Hence

concentrations to the solution, causes an increase in viscosity. Hence these are used as thickening agents, gelling agents, emulsifying agents, and stabilizers in the food industries. Also they are used in other industries as adhesives binding agents, clarifying agents, encapsulating agents, flocculating agents, swelling agents, foam stabilizers, .One of the natural gums is Locust bean gum. that it is very costly and difficult to extract. Locust bean gum (LBG, also known as carob gum) is a galactomannan[32] vegetable gum which is extracted from the

seeds of the carob tree. This tree is mostly found in the Mediterranean region. But the main problem associated with LGB is This problem is solved by the use of pullulanase enzyme on gaur galactomannan. Pullulanase enzymes remove galactose residues from the gaur galactomannan to produce modified gaur galactomannans [33]. These modified gaur galactomannans have all the functional properties of the locust bean gum and also have better rheological properties like viscosity and elasticity [34]. Pullulanase is relatively inexpensive, easily available and the modified guar galactomannan which is produced by galactomannan can be utilized for various food and non-food applications of the locust bean gum. International Journal of Science Innovations and Discoveries, Volume 2, Issue 2, March-April 2012 its action has enhanced rheological properties along with the functional properties of LGB .Hence these modified

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Nishank Prakash et al., IJSID, 2012, 2 (2), 266-273 g) In the production of resistant starch cannot be broken down by the enzymes present in the human small intestine .It is generally considered to be one of the most pullulanase enzyme.[36] Resistant starch is produced by the amylose retrogradation process amylose retrogradation [38]. Retrograded amylose yield is increased by the removal of amylopectin with the pullulanase.[38]
[37].

important components of the total dietary fibre (TDF) [35]. Resistant starch is produced by the hydrolysis of maize starch by also called asprebiotic fiber because it increases the growth of the healthy bacteria and also reduces the growth of the properties hence it keeps colon tissue healthy[42].

Resistant starch (RS) is that portion of the starch which is partially or wholly fermented in the large intestines. These

Resistant starch acts as a replacement for wheat products in the foods that are required to be gluten-free. Resistant starch is potentially harmful bacteria in the bowel[39]. The fermentation of the resistant starch in the intestines results in the reduction Butyrate acts as the primary source of energy for the colonic cells .also butyrate has anti-carcinogenic and anti inflammatory Pullulanase is a very potent enzyme in terms of its applications. The products obtained by the action of pullulanase CONCLUSION

Amylopectin interferes with

of intestinal pH, secondary resistant acids, ammonia and phenols[40]. Resistant starch also leads to the formation of butyrate[41].

have various pharmaceutical, neutraceutical properties. Pullulanase is cheap and easily available hence it helps in the economical production of various products like resistant starch and substitutes of LBG. The saccharification property of this advancement of technology it could be used in for various other applications which would directly or indirectly affect the humans. 1. 2. 3. 4. 5. 6. 7. 8. a REFERENCES Newly Isolated Thermophilic Anaerobic Bacterium Ven5, Applied production could also be done by the use of pullulanase thus providing a source of fuel. The basic property of breaking

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