Applications of Cellulases – Review R

IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES

Available online at www.ijpcbs.com Review Article

APPLICATIONS OF CELLULASES – REVIEW R. Sharada1*, G. Venkateswarlu2, S.Venkateswar3 and M. AnandRao4 1A.P.S.W.R.Junior College, Zaffargadh, Warangal, Andhra Pradesh, India. 2Bharat Heavy Electricals Limited, R.C.Puram, Hyderabad, Andhra Pradesh, India. 3University College of Technology, Osmania University, Hyderabad, Andhra Pradesh, India. 4University College of Sciences, Osmania University, Hyderabad, Andhra Pradesh, India.

ABSTRACT Cellulases are important enzymes not only for their potential applications in different industries like industries of animal feed, agricultural, laundry & detergent, textile, paper&pulp, wine &brewery, food processing, olive oil extraction. Carotenoid extraction, pharmaceutical & medical sciences, analytical applications, protoplast production, genetic engineering and pollution treatment, but also for the significant role in bio conversion of agriculture wastes into sugar and bio ethanol. This review paper simply assesses about: Enzymes, Cellulose, Cellulases& its types, Cellulase production and detailed study on Applications of cellulases.

Keywords: Enzymes, Cellulose, Cellulase.

INTRODUCTION hydrolases, the remaining 15% are divided Enzymes are biological catalysts which are the among oxidoreductases and isomerases. most remarkable, highly specialized and Enzymes are widely used in food, detergents, energized protein molecules found in every cell analytical applications, medical sciences, and are necessary for life. There are over 2000 production of chemicals & waste treatment. known enzymes, each of which is involved with one specific chemical reaction. These proteins Cellulose and their functions are determined by their Cellulose is found in plants as micro fibrils (2 – shape. They have extraordinary catalytic power, 20nm diameter and 100 – 40,000nm long). often for greater than that of synthetic or These form the structurally strong frame work inorganic catalysts. in the cell walls. Cellulose is mostly prepared Enzymes are vital to every biochemical process. from wood pulp. They have a high degree of specificity for their Cellulose is a linear glucose polymer composed substrates. They accelerate chemical reactions of anhydro glucose units coupled to each other tremendously and they function in aqueous by beta – 1,4 – glucosidic bonds and its most solutions under mild conditions and of normal abundant biological compound on terrestrial temperatures. earth. Enzymes like other proteins have molecular Cellulose is an insoluble molecule consisting of weights ranging from about 12,000 to more than residues between 2,000 – 14,000 with some of 1 million. Some enzymes require no chemical their properties being somewhat shorter. It groups for activity, but many require non- forms crystals (Cellulose Ia) where, intra protein co-enzymes or co-factors for their molecular and intra strand hydrogen bonds hold catalytic function. the network flat allowing the more hydrophobic According to International Union of Biochemists ribbon faces to the stack. Each residue is 1979, Six classes of enzymes have been oriented 1800to the next with the chain designated. They are: 1. Oxido – reductases, 2. synthesized two residues at a time. The natural Transferases, 3.Hydrolases, 4.Lyases, crystal is made up from meta stable“Cellulose I” 5.Isomerases, 6.Ligases. Major sources of with all the cellulose strands parallel and no industrial enzymes reveal that about 85% are inter sheet hydrogen bonding.

424 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

Cellulose has many uses as an anticake agent, 3. β – glucosidase / Cellobiase emulsifier, stabilizer, dispersing agent, thickener β– glucosidase ( E.C.3.2.1.21) completes the and gelling agent, but these are generally process of cellulose hydrolysis by cleaving subsidiary to its most important use of holding cellobiose and removing glucose from the non - on to water. Water cannot penetrate crystalline reducing ends of oligosaccharides. cellulose, but dry amorphous cellulose absorbs Complete degradation of cellulose to glucose water becoming soft and flexible. requires the synergistic action of all the three The degradation of cellulosic materials has components. gained increasing attention due to its world wide availability and immense potential for Cellulase production transforming them into sugars, fuels and Successful utilization of cellulosic materials as chemical feed stocks. renewable carbon sources is dependent on the Enzymatic hydrolysis hold tremendous promise development of economically feasible process due to the high specificity and production of technologies for cellulase production.It has been high yields of glucose without generation of calculated that cellulase production accounts for degradation products, unlike acid hydrolysis, ~27 – 20% of the cost of ethanol production utility cost are also low as hydrolysis occurs fromlignocellulosic materials. So, to keep the under mild reaction conditions. Microorganisms costs down, it is therefore very important to use that degrade cellulose are both abundant and a substrate that is inexpensive, such as wheat ubiquitous in nature. The enzymatic hydrolysis straw, rice bran, green gram husk etc.Large no. of cellulose requires the use of cellulase. of microorganisms are capable of degrading cellulose, fungi, bacteria are the main cellulase Cellulase producing microorganisms. Cellulase is a class of enzyme that catalyzes the cellulolysis i.e., hydrolysis of cellulose. Celulase Major steps involved in cellulase production is a multiple enzyme system consisting of endo –  Selection of potent strain as cellulose 1, 4 –β–D – glucanases and exo – 1, 4 –β– D – source. glucanases along with cellobiase (β– D –  Selection of waste as substrate, eg. glucosideglucano hydrolase). Lignocellulosic agricultural waste.  Enrichment of lignocellulosic waste in Types of Cellulases carbon content by process of pre- Fractionation studies on culture filtrate have treatment. demonstrated that, there are ‘three’ major types  Fermentation growth of selected strain of enzymes involved in the hydrolysis of native utilizing the pre-treated lignocellulosic cellulose to glucose, namely: waste. 1.Endo – glucanase  Harvesting the biomass cultivated after 2. exo – glucanase fermentation. 3. β– glucosidase / cellobiase.  Downstream processing of extra cellular enzymes and recovery of cellulase. 1. Endo – glucanase From the above steps, it can be observed that, Endo – glucanase (endo –β– 1, 4 – glucanase, there are three main stages for the production of endo –β– 1, 4 – D – glucan – 4 – cellulase. They are ‘pre - fermentative stage’, glucanohydrolase, E.C.3.2.1.4) ofen called CMC where the pre - treatment of substrate and ase, hydrolysis carboxyl methyl cellulose or medium preparationis done, followed by swollen cellulose, due to which there is a rapid ‘fermentative stage’ for cultivation of organism decrease in chain length along with a slow and finally ‘post – fermentative stage’ involving increase in reducing groups.Endoglucanse also downstream processing and product recovery. acts on cellodextrins, the intermediate products of cellulose hydrolysis and converts them to Cellulases Applications cellobiase and glucose. Recent development on bio – chemistry, genetics and protein as well as on the structure 2. Exo – glucanase function relationships of cellulases including Exoglucanase (Exo–β– 1, 4 – D – glucan – 4 – cellulosomes and related enzymes from bacteria cellobiohydrolase, E.C. 3.2.1.91) degrades and fungi has led to speculation and anticipation cellulose by splitting off the cellobiost units from of their enormous commercial potential in bio - the non- reducing end of the chain. technology and research.Cellulases have been a Cellobiohydrolase does not degrade cotton potential candidate for research by both rapidly, but can effect considerable academic and industrial research saccharification of micro crystalline celluloses. groups.Biotechnological conversion of cellulosic

425 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

biomasses potentially sustainable approach to chemical pretreatment by aerobic enzymatic develop novel bioprocesses and hydrolysis of chopped or milled biomass. products.Cellulase have become the focal biocatalysts due to their complex nature and Bio–Ethanol Industry wide spread industrial applications. Enzymatic saccharification of lignocellulosic Cellulases were initially investigated several materials such as rice bran, sugarcane bagasse, decades back for the bio conversion of the corncob, rice straw, green gram husk, saw dust, biomass which gave way to research in the and forest residues by cellulases for bioethanol industrial applications of the enzyme in Waste production is perhaps the most popular management, Bio – ethanol industry, Animal application currently being investigated. feed industry, Agricultural industry, Bioconversion of lignocellulosic materials into Laundry&detergent industry, Textile industry, useful and higher value products normally Paper& pulp industry, Wine& Breweryindustry, requires multistep processes. These processes Food processing industry, Olive oil extraction, include; pretreatment (mechanical, chemical, or Carotenoid extraction, Pharmaceutical & biological), hydrolysis of the polymers to medical sciences, Analytical applications, produce readily metabolizable molecules (e.g., Protoplast production, Genetic engineering and hexose and pentose sugars), bioconversion of Pollution treatment. these smaller molecules to support microbial growth and/or produce chemical products, and Waste Management the separation and purification of the desired Lignocellulose is the most abundant plant cell products. The utility cost of enzymatic wall component of the biosphere and the most hydrolysis may be low compared with acid or voluminous waste produced by our society. alkaline hydrolysis because enzyme hydrolysis Biomass is the only domestic, sustainable and is usually conducted at mild conditions (pH 4–6 renewable primary energy resource that can and temperature 45–50°C) and does not have provide liquid transportation fuels. The corrosion issues. conversion of cellulosic waste to useful by – Technologies are currently available for all steps products has long been recognized as a in the bioconversion of lignocellulosics to desirable endeavour. Disposal of cellulosic ethanol and other chemical products. However, municipal solid waste through processes that some of these technologies must be improved to would also desire energy production are ofthe produce renewable biofuel and other particular interest. The benefits would be ‘Two byproducts at prices, which can compete with fold’: Firstly, the amount of cellulose waste more conventional production systems. Not only would be diminished and its effects on our the recalcitrance of the substrate, but also environment will be reduced and Secondly, the several other factors that also limit cellulase pollutant would be converted to an alternative efficiency during the hydrolysis process source of energy to help displace our growing including end product inhibition, thermal dependence on fossil fuels. deactivation of the native protein, nonspecific The biological decomposition of organic matter binding to lignin , and irreversible adsorption of principally to methane and carbon dioxide by the enzymes to the heterogeneous substrate . ‘Anaerobic’ digestion is a natural process that To reduce the enzyme cost in the production of occurs readily in solid waste landfills. In natural fuel ethanol from lignocellulosic biomass, two anaerobic digestion processes some members of aspects are widely addressed: optimization of the microbial consortia collectively produce the cellulase production and development of a fermentable sugars from polysaccharides and more efficient cellulase-based catalysis system. others specialize in converting sugars to Protein engineering and directed evolution are methane and carbon dioxide.Such mixed powerful tools that can facilitate the fermentations are notoriously difficult to development of more efficient establish and maintain at large scale. The thermophiliccellulases . Strategies for recycling anaerobic biological conversion of the major and re-usage of the enzymes may also be used to polymerize components of solid waste requires reduce enzymatic hydrolysis costs. The recovery appropriate microorganisms and hydrolytic of enzymes is largely influenced by adsorption enzyme systems. Extra cellular hydrolytic of the enzymes onto the substrate, especially to enzymes such as cellulase and lipases have been lignin and enzyme inactivation. There are shown to be effective in the post hydrolysis of several reports where the nonspecific and anaerobic digester effluent solids. irreversible adsorption of cellulase to lignin has In conventional technologies for the ‘Aerobic’ been observed. Besides, there are also reports degradation of lignocellulose, lignocellulosic where the compounds that mimic cellulose or sugars are typically released by thermal the compounds have high affinity towards lignin

426 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

have been used to prevent the adsorption of Cellulases can be used to improve silage cellulases to lignin. Moreover, recently Scott and production for cattle feeding, which involves coworkers have filed a US patent enhancement of the digestibility of grasses (20100221778) on novel lignin-resistant containing large amounts of potentially total cellulase enzyme, in which linker peptides have digestible nutrients and energy values together been modified to prevent their adsorption onto with only small amounts of water-soluble lignin and enhance the enzyme activity. Among carbohydrates. The forage diet of ruminants, different strategies to recover and reuse the which contains cellulose, hemicellulose, pectin, cellulasesis concentration of the cellulose and lignin, is more complex than the cereal- fraction by ultrafiltration to remove sugars and based diet of poultry and pigs. Enzyme other small compounds that may inhibit the preparations containing high levels of cellulase, action of the enzymes and recycling of hemicellulase, and pectinase have been used to immobilized enzymes, which enables separation improve the nutritive quality of forages. of the enzymes from the process flow. However, Nevertheless, the results with the addition of the recycling techniques are mostly tested at enzyme preparations containing cellulase, laboratory scale. Therefore, the ability to scale hemicellulase and pectinase to ruminant diet up the techniques, the robustness, and are somewhat inconsistent. feasibility still needs to be demonstrated. Animal feedstock production processes generally include heat treatments that inactivate Animal feed Industry potential viral and microbial contaminants. Applications of cellulases and hemicellulases in Application of thermophiliccellulase in the animal feed industry have received feedstock production has the potential to reduce considerable attention because of their potential pathogens as well as to enhance digestibility and to improve feed value and performance of nutrition of the feed, thereby facilitating a animals. Pretreatment of agricultural silage and combination of heat treatment and feed grain feed by cellulases or xylanases can transformation in a single step. The cellulases improve its nutritional value. The enzymes can and hemicellulases are responsible for partial also eliminate antinutritional factors present in hydrolysis of lignocellulosic materials, dehulling the feed grains, degrade certain feed of cereal grains, hydrolysis of β-glucans, and constituents to improve the nutritional value, better emulsification and flexibility of feed and provide supplementary digestive enzymes materials, which results in the improvement in such as proteases, amylases, and glucanases. For the nutritional quality of animal feed. Moreover, instance, the dietary fiber consists of nonstarch these enzymes can cause partial hydrolysis of polysaccharides such as arabinoxylans, plant cell wall during silage and fodder cellulose, and many other plant components preservation. including resistant dextrins, inulin, lignin, The use of enzymes in animal nutrition became waxes, chitins, pectins, β-glucan, and more important after the prohibition of using oligosaccharides, which can act as anti- some nutritive ionophore antibiotics, which nutritional factor for several animals such as were previously used in the EU countries. There swine In this case, the cellulases effectively is a large variation in the digestibility of starch hydrolyse the anti-nutritional factor, cellulose, origins. The low digestibility of some starches in the feed materials into easily absorbent promotes the appearance of some digestive ingredient thus improve animal health and tract diseases because the non-digested and performance. non-absorbed starch reaching the large intestine β-Glucanases and xylanases have been used in can act as a substrate for bacterial fermentation the feed of monogastric animals to hydrolyze supporting the proliferation of some potentially nonstarch polysaccharides such as β-glucans hazardous pathogenic bacteria. Cellulases have a and arabinoxylans. Cellulases, used as feed positive effect on the caecal fermentation additives alone or with proteases, can processes by increasing the production of significantly improve the quality of pork meat. propionic acid, which act as a bacteriostatic Glucanases and xylanases reduce viscosity of material and thus can decrease the colonization high fibre rye- and barley-based feeds in poultry of pathogenic bacteria. and pig. These enzymes can also cause weight gain in chickens and piglets by improving Agricultural Industry digestion and absorption of feed materials. Various enzyme preparations consisting of Most low-quality feedstuffs contain higher different combinations of cellulases, concentrations of cellulose, small amounts of hemicellulases, and pectinases have potential protein and fat and relatively high ash contents applications in agriculture for enhancing growth when compared with high-quality feedstuffs. of crops and controlling plant diseases. Plant or

427 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

fungal protoplasts produced using microbial conventional detergent ingredients. Nowadays, hydrolases can be used to produce hybrid liquid laundry detergent containing anionic or strains with desirable properties. Cellulases and nonionic surfactant, citric acid or a water- related enzymes from certain fungi are capable soluble salt, protease, cellulose, and a mixture of of degrading the cell wall of plant pathogens in propanediol and boric acid or its derivative has controlling the plant disease. Fungal β- been used to improve the stability of cellulases. glucanases are capable of controlling diseases As most of the cellulose fibers in the modern by degrading cell walls of plant pathogens. Many textile industry enzymes are used increasingly cellulolytic fungi in the finishing of fabrics and clothes are including Trichoderma sp., Geocladium sp.,Chaet arranged as long, straight chains of some small omium sp., and Penicillium sp. are known to fibers can protrude from the yarn or fabric. The play a key role in agriculture by facilitating cellulases are applied to remove these rough enhanced seed germination, rapid plant growth protuberances for a smoother, glossier, and and flowering, improved root system and brighter-colored fabric. increased crop yields. Although these fungi have both direct (probably through growth- Textile Industry promoting diffusible factor) and indirect (by Cellulases are the most successful enzymes used controlling the plant disease and pathogens) in textile wet processing, especially finishing of effects on plants, it is not yet clear how these cellulose-based textiles, with the goal of fungi facilitate the improved plant performance. improved hand and appearance. Traditional It has been reported that β-1,3-glucanase and N- stonewashing of jeans involves amylase- acetyl-glucosaminidase from T. mediated removal of starch coating (desizing) harzianum strain P1 synergistically inhibited and treatment (abrasion) of jeans with pumice the spore germination and germ tube elongation stone (1-2 kg/pair of jeans) in large washing of B. cinerea . Moreover, the exoglucanase machines. Cellulases have been successfully promoters of Trichoderma are used for the used for the biostoning of jeans and biopolishing expression of the different proteins, enzymes, of cotton and other cellulosic fabrics. During the and antibodies in large amount. The biostoning process, cellulases act on the cotton exoglucanase promoters of Trichodermahave fabric and break off the small fiber ends on the been used for the expression of chymosin and yarn surface, thereby loosening the dye, which is other proteins: glucoamylase, lignin peroxidase, easily removed by mechanical abrasion in the and laccase. wash cycle. The advantages in the replacement Cellulases have also been used for the of pumice stones by a cellulose-based treatment improvement of the soil quality. Traditionally include less damage of fibers, increased straw incorporation is considered an important productivity of the machines, and less work- strategy to improve soil quality and reduce intensive and environment benign. dependence on mineral fertilizers. Many studies While the bio-polishing is usually carried out have attempted to hasten straw decomposition during the wet processing stages, which include via microbial routes. Cellulolytic fungi desizing, scouring, bleaching, dyeing, and applications such as Aspergillus, Chaetomium, finishing. The acidic cellulases improve softness and Trichoderma, and actinomycetes have and water absorbance property of fibres, shown promising results. Exogenouscellulase strongly reduce the tendency for pill formation, supplementation accelerated decomposition of and provide a cleaner surface structure with less cellulose in soil. Therefore, using exogenous fuzz. Cellulase preparations rich in cellulase may be a potential means to accelerate endoglucanases are best suited for biopolishing straw decomposition and increase soil fertility. enhancing fabric look, feel, and color without needing any chemical coating of fibers. The Laundry and Detergent Industry action of cellulases removes short fibers, surface Use of cellulases along with protease and lipase fuzziness, creates a smooth and glossy in the detergents is a more recent innovation in appearance, and improves color brightness, this industry. Cellulase preparations capable of hydrophilicity and moisture absorbance, and modifying cellulose fibrils can improve color environmentally friendly process. brightness, feel, and dirt removal from the Similarly, endoglucanase activity-rich cellulase cotton blend garments. The industrial is also proved better for biofinishing. Most application of alkaline cellulases as a potential cotton or cotton-blended garments, during detergent additive is being actively pursued repeated washing, tend to become fluffy and with a view to selectively contact the cellulose dull, which is mainly due to the presence of within the interior of fibers and remove soil in partially detached microfibrils on the surface of the interfibril spaces in the presence of the more garments. The use of cellulases can remove

428 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

these microfibrils and restore a smooth surface that improvements in dewatering and deinking and original color to the garments. The use of of various pulps result in the peeling of the cellulase also helps in softening the garments individual fibrils and bundles, which have high and in removal of dirt particles trapped within affinity for the surrounding water and ink the microfibril network. particles. The main advantages of enzymatic Interestingly, there are several reports where deinking are reduced or eliminated alkali usage, the performance of the whole cellulase improved fiber brightness, enhanced strength preparations was quite different from the properties, higher pulp freeness and cleanliness, enzyme rich in endoglucanase activity, and that and reduced fine particles in the pulp. Moreover, the latter offered better performance in deinking using enzymes at acidic pH also applications where losses in fabric strength and prevents the alkaline yellowing, simplifies the weight were minimum. Depilling/cleaning deinking process, changes the ink particle size and/or ageing effects are the result of the distribution, and reduces the environmental synergistic action of cellulases and mechanical pollution. Although enzymatic deinking can action, simultaneously or sequentially. Attempts lower the need for deinking chemicals and have also been made via cellulases treatment to reduce the adverse environmental impacts of improve the dimensional stability of cellulosic the paper industry, the excessive use of enzymes fabrics and to upgrade the surface and dyeing must be avoided, because significant hydrolysis properties of bleached cotton, mercerized of the fines could reduce the bondability of the cotton, and cotton/polyester blend fabric fibers. (50/50), using the pad-wet batch technique, Interestingly, the use of cellulases in improving followed by subsequent washing under the drainage has also been pursued by several mechanical action. mills with the objective to increase the production rate. Enzyme treatments remove Paper & Pulp Industry some of the fines or peel off fibrils on the fiber Interest in the application of cellulases in the surface and dissolved and colloidal substances, pulp and paper industry has increased which often cause severe drainage problems in considerably during the last decade. The paper mills. In this aspect, cellulases have shown mechanical pulping processes such as refining considerable improvement in the overall and grinding of the woody raw material lead to performance of paper mills. Enzymatic pulps with high content of fines, bulk, and treatment also destabilizes the lipophilic stiffness. While in contrast, biomechanical extractives in the filtrates and facilitates their pulping using cellulases resulted in substantial attachment to thermo-mechanical pulping energy savings (20–40%) during refining and fibers. These enzymes are also used in improvements in hand-sheet strength preparation of easily biodegradable cardboard, properties. manufacturing of soft paper including paper Mixtures of cellulases (endoglucanases I and II) towels and sanitary paper and removal of and hemicellulases have also been used for adhered paper. biomodification of fiber properties with the aim of improving drainage and beatability in the Wine and Brewery Industry paper mills before or after beating of pulp. Microbial glucanases and related Mansfield et al. studied the action of a polysaccharides play important roles in commercial cellulase preparation on different fermentation processes to produce alcoholic fractions of Douglas fir kraft pulp and observed beverages including beers and wines. These that the cellulase treatment decreased the enzymes can improve both quality and yields of defibrillation reducing the fibre coarseness. the fermented products .Glucanases are added While endoglucanases have the ability to either during mashing or primary fermentation decrease the pulp viscosity with a lower degree to hydrolyze glucan, reduce the viscosity of of hydrolysis, cellulases have also been reported wort, and improve the filterability. to enhance the bleachability of softwood kraft In wine production, enzymes such as pectinases, pulp producing a final brightness score glucanases, and hemicellulases play an comparable to that of xylanase treatment. important role by improving color extraction, Cellulases alone, or used in combination with skin maceration, must clarification, filtration, xylanases, are beneficial for deinking of different and finally the wine quality and stability. β- types of paper wastes. Most applications Glucosidases can improve the aroma of wines by proposed so far use cellulases and modifying glycosylated precursors. Macerating hemicellulases for the release of ink from the enzymes also improve pressability, settling, and fiber surface by partial hydrolysis of juice yields of grapes used for wine carbohydrate molecules. It has been postulated fermentation. A number of commercial enzyme

429 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

preparations are now available to the wine infusion of enzymes such as pectinases and β- industry. The main benefits of using these glucosidases. Enzyme mixtures containing enzymes during wine making include better pectinases, cellulases, and hemicellulases are maceration, improved color extraction, easy also used for improved extraction of olive oil. clarification, easy filtration, improved wine Use of macerating enzymes not only improves quality, and improved stability. the cloud stability and texture of nectars and Beer brewing is based on the action of enzymes purees, but also decreases their viscosity activated during malting and fermentation. rapidly. Thus, the macerating enzymes, Malting of barley depends on seed germination, composed of mainly cellulase and pectinase, which initiates the biosynthesis and activation play a key role in food biotechnology, and their of α- and β-amylases, carboxypeptidase, and β- demand will likely increase for extraction of glucanase that hydrolyze the seed reserves. In juice from a wide range of fruits and vegetables. an earlier study, researchers observed that Furthermore, infusion of pectinases and β- endoglucanase II and exoglucanase II of glucosidases has also shown to alter the texture, the Trichoderma cellulase system were flavor, and other sensory properties such as responsible for a maximum reduction in the aroma and volatile characteristics of fruits and degree of polymerization and wort viscosity. vegetables. Significant and reproducible improvements in grape pressability, settling rate, and total juice Olive Oil extraction yield were achieved using a combination of In recent years, extraction of olive oil has macerating enzymes. Such improvements were attracted the interest of international market noticeable only with a correct balance of because of its numerous health claims. pectinases, cellulases, and hemicellulases. Using Extraction of olive oil involves (1) crushing and three varieties (Soave, Chardonnay, and grinding of olives in a stone or hammer mill; (2) Sauvignon) of white grapes,reports show that, passing the minced olive paste through a series the performance of Cytolase 219 (mixture of of malaxeurs and horizontal decanters; (3) high- cellulase, pectinase, and xylanase) in wine speed centrifugation to recover the oil. To making and 10–35% increase in the extraction produce high-quality olive oil, freshly picked, of the first wine must, a 70–80% increase in the clean, and slightly immature fruits have been must filtration rate, 50–120 minutes decrease in used under cold pressing conditions. However, pressing time, 30–70% decrease in must high yields have been obtained with fully viscosity, 20–40% saving of energy during ripened fruit, when processed at higher than cooling of fermenter, and a significant ambient temperatures, but this resulted in oil improvement in wine stability. A range of with high acidity, rancidity, and poor aroma. improved enzymes like cellulase and pectinase Hence, an improved method for the extraction of that would be exogenously added to the process high-quality olive oil was needed to meet the are expected to enhance the productivity of growing consumer demand. The commercial existing brewing processes in future. enzyme preparation, Olivex (a pectinase preparation with cellulase and hemicellulase Food processing industry from Aspergillusaculeatus), was the first Cellulases have a wide range of potential enzyme mixture used to improve the extraction applications in food biotechnology as well. The of olive oil. Furthermore, the use of macerating production of fruit and vegetable juices requires enzymes increased the antioxidants in improved methods for extraction, clarification, extravirgin olive oil and reduced the induction and stabilization. Cellulases also have an of rancidity. The main advantages of using important application as a part of macerating macerating enzymes during olive oil extraction enzymes complex (cellulases, xylanases, and are (1) increased extraction (up to 2 kg oil per pectinases) used for extraction and clarification 100 kg olives) under cold processing conditions; of fruit and vegetable juices to increase the yield (2) better centrifugal fractionation of the oily of juices. The use of macerating enzymes must; (3) oil with high levels of antioxidants and increases both yield and process performance vitamin E; (4) slow induction of rancidity; (5) without additional capital investment. The overall improvement in plant efficiency; (6) low macerating enzymes are used to improve cloud oil content in the waste water. Likewise, the stability and texture and decrease viscosity of macerating enzymes could play a prominent the nectars and purees from tropical fruits such role in the extraction of oils from other as mango, peach, papaya, plum, apricot, and agricultural oilseed crops. pear. Texture, flavor, and aroma properties of These enzymes can also be used during olive fruits and vegetables can be improved by paste malaxation. The presence of collateral reducing excessive bitterness of citrus fruits by activities of a cellulase and hemicellulase nature

430 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

of the enzymatic formulation guarantees a rapid ‘malabsorption’.Since, humans poorly digest and intense disintegration of the cell walls and cellulose fiber, taking a digestive enzyme membranes of the olive fruits, thereby favoring product, like Digestin, that contains cellulase the passage of noble substances (particularly enzymes is not only necessary, but also vital for the polyphenols and aromatic precursors) into healthy cells. the final product. It is also used to decrease olive paste viscosity in olive oil production and to Analytical Applications intensify the process of extracting the Use of cellulases in various aspects of chemical polyphenolic substances contained in the olive analysis has been a growing interest. The fruit. It is necessary to underline that the principal analytical application of enzymes in selected enzymes are naturally present inside the diagnostics and food analysis. the olive fruit, but they are strongly deactivated during the critical pressing step, probably Protoplast Production because of oxidation phenomena. So, the For overcoming interspecific, incompatibility, replacement of these enzymes is expected to be protoplast combination method is a proper appropriate in relation with the role they play in procedure for making a new plant with desired determining the final product quality. triats. For this purpose, protoplast preparation is a first and important step. Hence, Carotenoid Extraction experimental results show that protoplast yield Carotenoids are the main group of coloring was significantly affected to the maximum substances in nature being responsible for many extent by the enzyme cellulase along with plant colors from red to yellow. There is a pectinase. continuously growing market for carotenoids as food colorants due to their desirable properties, Genetic Engineering such as their natural origin, null toxicity, and Combining different types of enzymes and high versatility, providing both lipo- and genetically engineering new enzymes that work hydrosoluble colorants with colors ranging from together to release both hemicellulosic sugars yellow to red. In addition, provitaminA activity, and cellulosic sugars. Biotechnological advances, a role in lipid oxidation, and anticarcinogenic especially in genetic engineering of properties are very important biological commercially useful enzyme producing strains functions of these pigments. are playing an important role in solving some of Usually a combination of cellulolytic and the bottlenecks that occur in production and will pectinolytic enzymes accelerates the rate of be vital in securing cellulosic ethanol as a hydrolysis for achieving complete liquefaction. competive liquid fuel for the future by using Cellulase randomly splits cellulose chains into cellulase. glucose whereas commercial pectinase preparations from Aspergillusniger have Pollution Treatment pectinesterase (PE), polygalacturonase (PG), Environmental pollution is growing more & and pectin lyase (PL) activity. The use of more due to the indiscriminate and frequently pectinase and cellulase enzymes disrupts the deliberate release of hazardous, harmful cell wall of orange peel, sweet potato and carrot, substances. Research efforts have been devoted and releases the carotenoids in the chloroplasts to develop new, low cost, low technology, eco- and in cell fluids. These pigments remain in their friendly treatments capable of reducing and natural state still bound with proteins. This even eliminating pollution in the atmosphere, bonded structure prevents pigment oxidation the hydrosphere and soil environments.Among and also affects color stability, whereas solvent biological agents, enzymes have a great extraction dissociates the pigments from the potentiality to effectively transform and detoxify proteins and causes water insolubility and ease polluting substances because they have been of oxidation. recognized to be able to transform pollutants a detectable rate are potentially suitable to Pharmaceutical& Medical Sciences restore polluted environments. Enzymes are Considerable research in this area is being done capable of transforming some classes of and newer applications are being discovered, pollutants into innocuous products. such as the treatment of enzyme defiencies. Several substances with high polluting potential Enzymes are also used as anti- tumor or anti - are present in the environment & affect soil, microbial agents, treatment of blood cloth and sediments, water, air, microbial organisms, herniated discs. Effect of enzyme cellulaseis as plants, animals & humans. The origins and digestive aid i.e. celulase digests fiber. It helps in sources of pollution are different, industrial the remedy of digestive problems such as activities such as mining and metal processing,

431 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

petrochemical and industrial complexes, ACKNOWLEDGEMENTS industry effluent, chemical weapons production, We the authors wish to thank pulp &paper industries, dye industries, SravaniVidhushiGogu and MeghanaShivaniGogu anthropogenic activities such as traffic, for their friendly and presentational support. agricultural practices and others. Pollutants may affect the health of humans, animals and REFERENCES environments for several causes. 1. Sharada R, Swathi K, Venkateswar S, Therefore, a growing interest is being devoted AnandRao M and Narsi Reddy M. to the search of effective remediation Optimization of cultural conditions for technologies for partial or total recovery of cellulase production by polluted sites. Several methodologies have been Aspergillusniger (MTCC 2196) using applied for remediation of polluted systems, of submerged fermentation. Biosciences which ‘two’ basic strategies have been utilized: Biotechnology Research Asia. engineering and biological ones. Bioremediation 2009;6(1):289-292. appear now as appealing technologies being 2. Sharada R, Venkateshwarlu G, based on their enzymatic set. NarsiReddy M, Venkateswar S and The use of enzymatic proteins may represent a Anand Rao M. Production of Cellulase good alternative for overcoming most by Solid State Fermentation. IJPRD. disadvantages related to the use of 2012;4(4):224-230. microorganisms. The most representative 3. Sharada R, Venkateshwarlu G, enzymatic classes in the remediation of polluted Venkateswar S and Anand Rao M. environments are oxidoreductases, hydrolases Production of Cellulase – A Review. i.e., Cellulases and amylases. IJCBS. 2013;3(4):1070-1090. 4. Anupama and Ravindra P. Value CONCLUSION added food: Single cell protein. It was estimated that, the world sale of Biotechnol Adv. 2000;18(6): 459-479. industrial enzymes have already reached a 5. Anuppama and Ravindra P. Studies on market value of 1.6 billion US of which cellulase production of single cell protein by and allied enzymes occupy a significant position. Aspergillusniger insolid state Microbes are an attractive topic of interest for fermentation of rice bran. Braz Arch the production of cellulases and hemicellulases BiolTech. 2001a;44:79-88. due to their immense potential for cellulase 6. Edward A Bayer, Raphael Lamed and production, enzyme complexity and extreme Micheal E Himmel. The Potential of habitat variability. Microbial cellulases are Cellulases and Cellulosomes for preferred for their vast industrial applicability Cellulosic Waste Management. Current and relatively lower cost of production. In fact Opinion in Biotechnology. the craze for these cellulaseenzymes is 2007;18:237-245. increasing day to day worldwide for their use in 7. Ajoy K Sarkar and Nolan Etters J. waste management, food processing, Enzymatic hydrolysis of cotton pharmaceuticals,pulp&paper and other fibers:Modeling using an empirical industries. More and more research works are equation. The Journal of cotton resulting into improved scientific knowledge science. 2004;8:254-260. along with the success of meeting the growing 8. ArturCavaco-Paulo. Mechanism of demands of the cellulase and related enzymes cellulase action in textile processes. for generation of environment friendly textiles, Carbohydrate polymers. 1988;37:273- detergents, bio-pulping and bio-alcohols. 277. Moreover, it is opening new avenues for 9. Braz J Demuner, Nei Pereira Junior and utilization of various agro-wastes and organic Adelaide MS Antunes. Technology pollutants as a source of renewable prospecting on enzymes for the pulp energyinstead of dumping them to cause and paper industry. J Technol Manag environmental degradation. In near future Innov. 2011;6(3). newer knowledge of excellent cellulolytic and 10. William R Kenealy and Thomas W hemi-cellulolytic systems and adoption of Jeffries. Enzyme processes for pulp different biotechnological strategies will and paper: A review of recent certainlybringsgreat prospect in the field of developments.Chapter 12. industrial greenchemistry. 11. Ramesh Chander Kuhad, Rishi Gupta and Ajay Singh. Microbial cellulases and their industrial applications. Enzyme research. 2011;10.

432 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

12. Mai C, Kües U and Militz H. Lantana camara (red sage): Biotechnology in the wood industry. pretreatment, saccharification and Applied Microbiology and fermentation. Bioresource Technology. Biotechnology. 2004;63:477–494. 2010;101(21):8348–8354. 13. Singh A, Kuhad RC and Ward OP. 24. Stork G and Puls J. Changes in Industrial application of microbial properties of different recycled pulps cellulases, in Lignocellulose by endoglucanase treatments. Biotechnology. Future Prospects. InProceedings of the International 2007;345–358. Conference on Biotechnology in the 14. Akhtar M. Biochemical pulping of Pulp and Paper Industry: Recent aspen wood chips with three strains Advances in Applied and Fundamental of Ceriporiopsissubvermispora. Research. E. Srebotnik and K. Mesner. Holzforschung. 1994;48:199–202. 1996;1:145–150. 15. Pere J, Puolakka A, Nousiainen and 25. Karnis A. The role of latent and Buchert J. Action of delatent mechanical pulp fines in sheet purified Trichodermareesei cellulases structure and pulp properties. on cotton fibers and yarn. Journal of PaperiJaPuu-Paper Timber. Biotechnology. 2001;89(2-3):247–255. 1995;77:491–497. 16. Bhat MK. Cellulases and related 26. Kantelinen A, Jokinen O and Sarkki ML. enzymes in biotechnology. Effects of enzymes on the stability of Biotechnology Advances. 2000;18 colloidal pitch. InProceedings of the ( 5):355–383. 8th International Symposium on Wood 17. Dienes D, Egyházi A and Réczey K. and Pulping Chemistry. 1995;1:605– Treatment of recycled fiber 612. with Trichoderma cellulases. ndustrial 27. Buchert J, Oksanen T, Pere J, Siika-aho Crops and Products. 2004;20(1):11– M, Suurnakki A and Viikari L. 21. Applications of 18. Mansfield SD, Wong KKY, De Jong E Trichodermareesei enzymes in the and Saddler JN. Modification of pulp and paper industry Douglas-fir mechanical and kraft pulps in Trichoderma & Gliocladium- by enzyme treatment. Tappi Journal. Enzymes, G. F. Harman and C. P. 1996;79(8):125–132. Kubicek. of Biological Control and 19. Pere J, Siika-aho M, Buchert J and Commercial Applications. 1998;2:343– Viikari L. Effects of purified T. 363. reesei cellulases on the fiber 28. Salonen SM. Method for manufacturing properties of kraft pulp. Tappi Journal. paper or cardboard and product 1995;78(6):71–78. containing cellulose. US patent 20. Suominen P and Reinikainen T. 4980023, 1993. Foundation for biotechnical and 29. Hsu JC and Lakhani NN. Method of industrial fermentation research. making absorbed tissue from recycled InProceedings of the 2nd Symposium waste paper. US patent 6413363, on TrichodermaReesei Cellulases and 2002. Other Hydrolases (TRICEL '93). 30. Sharyo M, Sakaguchi H and Ohishi M. 1993;8, Espoo, Finland. Method of making sanitary paper from 21. ChanderKuhad R, Mehta G, Gupta R chemical pulp using a single and Sharma KK. Fed batch component cellulase that does not enzymaticsaccharification of contain cellulose-building domain. US newspaper cellulosics improves the patent 6468391, 1978. sugar content in the hydrolysates and 31. Hebeish A and Ibrahim NA. The impact eventually the ethanol fermentation of fronteir sciences on textile industry. by Saccharomyces cerevisiae. Biomass Colourage. 2007;54:41–55. and Bioenergy. 2010;34(8):1189– 32. Karmakar M and Ray RR. Current 1194. trends in research and application of 22. Kibblewhite RP, Bawden AD and microbialcellulases. Research Journal Brindley CL. TMP fiber and fines of Microbiology. 2011;6(1):41–53. qualities of 13 radiata pine wood 33. Uhlig H. Industrial Enzymes and Their types. APPITA. 1995;48:367–377. Applications. John Wiley& Sons, New 23. Kuhad RC, Gupta R, Khasa YP and York, NY, USA, 1998. Singh A. Bioethanol production from

433 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

34. Galante YM, DeConti A and Applied Microbiology. 1995;39:295– Monteverdi R. Application 333. of Trichoderma enzymes in food and 44. Wyman CE, Dale BE, Elander RT, feed industriesin Trichoderma and Holtzapple M, Ladisch MR and Lee YY. Gliocladium Enzymes, G. F. Harman Coordinated development of leading and C. P. Kubicek, Eds of Biological biomass pretreatment technologies. Control and Commercial Applications, Bioresource Technology. Taylor & Francis, London, UK, 2005;96(18):1959–1966. 1998;2:311–326. 45. Kuhad RC and Singh A. Lignocellulose 35. Sreenath HK, Shah AB, Yang VW, biotechnology: current and future Gharia MM and Jeffries TW. Enzymatic prospects. Critical Reviews in polishing of jute/cotton blended Biotechnology. 1993;13(2):151–172. fabrics. Journal of Fermentation and 46. Mosier N, Wyman C and Dale B. Bioengineering. 1996;81(1):18–20. Features of promising technologies for 36. Ibrahim NA, El-Badry K, Eid BM and pretreatment of lignocellulosic Hassan TM. A new approach biomass. Bioresource Technology. forbiofinishing of cellulose-containing 2005;96(6):673–686. fabrics using acid cellulases. 47. Yang and Wyman CE. Effect of xylan Carbohydrate Polymers. and lignin removal by batch and flow 2001;83(1):116–121. through pretreatment on the 37. Baker JO, McCarley JR and Lovett R. enzymatic digestibility of corn stover Catalytically enhanced endocellulase cellulose. Biotechnology and Cel5A from Bioengineering. 2004; 86(1):88–95. Acidothermuscellulolyticus. Applied 48. Taniguchi M, Suzuki H, Watanabe D, Biochemistry and Biotechnology A. Sakai K, Hoshino K and Tanaka T. 2005;121(1–3):129–148. Evaluation of pretreatment 38. Cortez JM, Ellis J and Bishop DP. Using with Pleurotusostreatus for enzymatic cellulases to improve the dimensional hydrolysis of rice straw. Journal of stability of cellulosic fabrics. Textile Bioscience and Bioengineering. Research Journal. 2002;72(8):673– 2005;100(6):637–643. 680. 49. Lee D, Yu AHC and Saddler JN. 39. Sukumaran RK, Singhania RR and Evaluation of cellulase recycling Pandey A. Microbial cellulases strategies for the hydrolysis of production, applications and lignocellulosic substrates. challenges. Journal of Scientific and Biotechnology and Bioengineering. Industrial Research. 2005;64(11):832– 1995;45(4):328–336. 844. 50. Singh A, Kumar PKR and Schugerl K. 40. Kuhad RC, Gupta R and Khasa YP. Adsorption and reuse of cellulases Bioethanol production from during saccharification of cellulosic lignocellulosic biomass: an overview materials. Journal of Biotechnology. in Wealth from Waste. B. Lal, Ed., Teri 1991;18(3):205–212. Press, New Delhi, India, 2010. 51. Bernardez TD, Lyford K, Hogsett DA 41. Kuhad RC, Singh A and Eriksson KE. and Lynd LR. Adsorption Microorganisms and enzymes involved of Clostridium thermocellumcellulases in the degradation of plant fiber cell onto pretreated mixed hardwood, walls. Advances in Biochemical Avicel, and lignin. Biotechnology and Engineering Biotechnology. Bioengineering. 1993;2(7):899–907. 1997;57:45–125. 52. Yang B and Wyman CE. Effect of xylan 42. Gupta R, Khasa YP and Kuhad RC. and lignin removal by batch and Evaluation of pretreatment methods in flowthrough pretreatment on the improving the enzymatic enzymatic digestibility of corn stover saccharification of cellulosic materials. cellulose. Biotechnology and Carbohydrate Polymers. 2011;84: Bioengineering. 2004;86(1):88–95. 1103–1109. 53. Yang B and Wyman CE. BSA treatment 43. Ghosh P and Singh A. Physicochemical to enhance enzymatic hydrolysis of and biological treatments for cellulose in lignin containing enzymatic/microbial conversion of substrates. Biotechnology and lignocellulosic biomass. Advances in Bioengineering. 2006;94(4):611–617.

434 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

54. Kumar R and Wyman CE. Effect of pretreatment. Journal of Membrane additives on the digestibility of corn Science. 2004;228(2):179–186. stover solids following pretreatment 65. Rai P, Majumdar GC, Das Gupta S and by leading technologies. Biotechnology De S. Effect of various pretreatment and Bioengineering. methods on permeate flux and quality 2009;102(6):1544–1557. during ultrafiltration of mosambi juice. 55. Scott BR, St-Pierre P, Lavigne J, Masri Journal of Food Engineering. N, White TC and Tomashek JJ. Novel 2007;78(2):561–568. lignin-resistant cellulase enzymes. US 66. Grassin C and Fauquembergue P. Fruit patent: 20100221778, 2010. juices in Industrial Enzymology. T. 56. Tu M, Chandra RP and Saddler JN. Godfrey and S. West, Eds., 226–264, Evaluating the distribution of MacMillan Press, London, UK, 2nd cellulases and the recycling of free edition, 1996. cellulases during the hydrolysis of 67. Humpf HU and Schreier P. Bound lignocellulosic substrates. aroma compounds from the fruit and Biotechnology Progress. the leaves of blackberry 2007;23(2):398–406. (Rubuslaciniata, L.). Journal of 57. Dourado F, Bastos M, Mota M and Agricultural and Food Chemistry. Gama FM. Studies on the properties of 1991;39(10):1830–1832 Celluclast/Eudragit L-100 conjugate. 68. Marlatt C, Ho CT and Chien M. Studies Journal of Biotechnology. of aroma constituents bound as 2002;99(2):121–131. glycosides in tomato. Journal of 58. Bamforth CW. Current perspectives on Agricultural and Food Chemistry. the role of enzymes in brewing. Journal 1992;40(2):249–252. of Cereal Science. 2009;50(3):353– 69. Dhiman TR, Zaman MS, Gimenez RR, 357. Walters JL and Treacher R. 59. Canales AM, Garza R, Sierra JA and Performance of dairy cows fed forage Arnold R. The application of beta- treated with fibrolytic enzymes prior glucanase with additional side to feeding. Animal Feed Science and activities in brewing. MBAA Technical Technology. 2002;101(1–4):115–125. Quarterly. 1998;25:27–31. 70. Godfrey T and West S. Textiles 60. Oksanen J, Ahvenainen J and Home S. in Industrial Enzymology. Macmillan Microbial cellulase for improving Press, London, UK, 2nd edition, filterability of wort and beer. 1996;360-371. In Proceedings of the 20th European 71. Ali S, Hall J and Soole KL. Targeted Brewery Chemistry Congress. expression of microbial cellulases in 1985;419–425, Helsinki, Finland. transgenic animals inCarbohydrate 61. Minussi RC, Pastore GM and Duran N. Bioengineering, S. B. Petersen, B. Potential applications of laccase in the Svensson, and S. Pedersen, Eds., vol. 10 food industry. Trends in Food Science of Progress in Biotechnology. Elsevier, and Technology. 2002;13(6-7):205– Amsterdam, The Netherlands, 216. 1995;279–293. 62. De Carvalho LMJ, de Castro IM and da 72. Shrivastava B, Thakur S, Khasa YP, Silva CAB. A study of retention of Gupte A, Puniya AK and Kuhad RC. sugars in the process of clarification of White-rot fungal conversion of wheat pineapple juice (Ananascomosus, L. straw to energy rich cattle feed. Merril) by micro- and ultra-filtration. Biodegradation. 2011;22(4):823–831. Journal of Food Engineering. 73. Graham H and Balnave D. Dietary 2008;87(4):447–454. enzymes for increasing energy 63. Baker RA and Wicker L. Current and availability in Biotechnology in Animal potential applications of enzyme Feeds and Animal Feedings, R. J. infusion in the food industry. Trends in Wallace and A. Chesson, Eds., VHC, Food Science and Technology. Weinheim, Germany. 1995;296–309. 1996;7(9):279–284. 74. Lewis GE, Hunt CW, Sanchez WK, 64. Youn KS,. Hong JH, Bae DH, Kim SJ and Treacher R, Pritchard GT and Feng P. Kim SD. Effective clarifying process of Effect of direct-fed fibrolytic enzymes reconstituted apple juice using on the digestive characteristics of a membrane filtration with filter-aid forage-based diet fed to beef steers.

435 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

Journal of Animal Science. activity of a glucan 1,3-beta- 1996;74(12):3020–3028. glucosidase and an N-acetyl-beta- 75. Paulo AC and Gubitz GM. Textile glucosaminidasefromTrichodermaharz Processing with Enzymes, Woodhead, ianum,” Phytopathology. Cambridge, UK, 2003. 1994;84(4):398–405. 76. Cowan WD. Animal feed in Industrial 85. Harkki A, Uusitalo J, Bailey M, Penttila Enzymology, T. Godfrey and S. West, M and Knowles JKC. A novel fungal Eds., 360–371, Macmillan Press, expression system: secretion of active London, UK, 2nd edition, 1996. calf chymosin from the filamentous 77. Pazarlioglu NK, Sariişik M and fungus Trichodermareesei. Telefoncu A. Treating denim fabrics Biotechnology. 1989;7(6):596–603. with immobilized commercial 86. Penttila M. Heterologous protein cellulasesProcess Biochemistry. production in Trichodermain 2005;40(2):767–771. Trichoderma & Gliocladium-Enzymes, 78. Pascual JJ. Recent advances on early G. F. Harman and C. P. Kubicek, Eds. weaning and nutrition around Biological Control and Commercial weaning. In Proceedings of the 2nd Applications. Taylor & Francis, Meeting of COST 848 Working Groups. London, UK, 1998;2:365-382. 2001;4:31–36, Hungary. 87. Saloheimo M and Niku-Paavola ML. 79. Fortun-Lamothe L, Gidenne T, Debray Heterologous production of a L and Chalaye F. Intake regulation, ligninolytic enzyme: expression of performances and health status the Phlebiaradiata laccase gene according to feeding strategy around in Trichoderma reesei. Biotechnology. weaning in Proceedings of the Second 1991;9(10):987–990. Meeting of COST 848 Working Group 88. Saloheimo M, Barajas V, Niku-Paavola 4, 40–41, Hungary, 2001. ML and Knowles JKC. A lignin 80. Chet I, Benhamou N and Haran S. peroxidase-encoding cDNA from the Mycoparatism and lytic enzymes white-rot fungus Phlebiaradiata: in Trichoderma and Gliocladium— characterization and expression Enzymes, G. F. Harman and C. P. in Trichodermareesei. Gene. 1989; Kubicek, Eds. Biological Control and 85(2):343–351. Commercial Applications. Taylor & 89. Ortiz Escobar ME and Hue NV. Francis, London, UK. 1998;2:327–342. Temporal changes of selected chemical 81. Bailey BA and Lumsden RD. properties in three manure—amended Directefects soils of Hawaii. Bioresource of Trichoderma and Gliocladium on Technology. 2008;99(18):8649–8654. plant growth and resistance to 90. Tejada M, Gonzalez JL, García-Martínez pathogens in Trichoderma & AM and Parrado J. Application of a Gliocladium—Enzymes, G. F. Harman green manure and green manure and C. P. Kubicek, Eds. Biological composted with beet vinasse on soil Control and Commercial Applications. restoration. effects on soil properties. Taylor & Francis, London, UK, Bioresource Technology. 1998;2:327–342. 2008;99(11):4949–4957. 82. Harman GE and Björkman T. Potential 91. Bowen RM and Harper SHT. and existing uses Decomposition of wheat straw and of Trichoderma and Gliocladium for related compounds by fungi isolated plant disease control and plant growth from straw in arable soils. Soil Biology enhancementin Trichoderma and and Biochemistry. 1990;22(3):393– Gliocladium, C. P. Kubicek and G. E. 399. Harman, Eds. Taylor and Francis, 92. Tiwari VN, Pathak AN and Lehri LK. London, UK, 1998;2:229–265. Effect of plant waste incorporation by 83. Harman GE and Kubicek CP. different methods under uninoculated Trichoderma and Gliocladium: and inoculated conditions on wheat Enzymes. Biological Control and crops. Biological Wastes. 1987;21(4): Commercial Applications, Taylor & 267–273. Francis, London, UK, 1998;2. 93. Abdulla HM and El-Shatoury SA. 84. Lorito M, Hayes CK, Di Pietro A, Woo Actinomycetes in rice straw SL and Harman GE. Purification, decomposition Waste Management. characterization, and synergistic 2007;27(6):850–853.

436 IJPCBS 2014, 4(2), 424-437 Sharada et al. ISSN: 2249-9504

94. Fontaine S, Bardoux G, Benest D, new vegetable enzyme extract during Verdier B, Mariotti A and Abbadie L. processing. European Food Research Mechanisms of the priming effect in a and Technology. 2003;216(2):109– Savannah Soil amended with cellulose. 115. Soil Science Society of America Journal. 99. Chiacchierini E, Mele G, Restuccia D 2004;68(1):125–131. and Vinci G. Impact evaluation of 95. Han W and He M. The application of innovative and sustainable extraction exogenous cellulase to improve soil technologies on olive oil quality. fertility and plant growth due to Trends in Food Science and acceleration of straw decomposition. Technology. 2007;18(6):299–305. Bioresource Technology. 100. Çinar I. Effects of cellulase and 2010;101(10): 3724–3731. pectinase concentrations on the colour 96. De Faveri D, Aliakbarian B, Avogadro yield of enzyme extracted plant M, Perego P and Converti A. carotenoids. Process Biochemistry. Improvement of olive oil phenolics 2005;40(2):945–949. content by means of enzyme 101. Ory R and Angelo AJSt. Enzymes in formulations: effect of different Food and Beverage Processing, enzyme activities and levels. American Chemical Society, Biochemical Engineering Journal. Washington, DC, USA, 1977. 2000;41(2):149–156. 102. Fennema OW. Food Chemistry, Marcel 97. Fantozzi P, Petruccioli G and Dekker, New York, NY, USA, 1985. Montedoro G. Trattamenti con additive 103. Bassi R, Pineau B, Dainese P and enzimaticialle paste di Marquardt J. Carotenoid-binding olivasottoposteadestrazione per proteins of photosystem II. European pressioneunica: influenze delle Journal of Biochemistry. cultivars, dell'epoca di raccolta e 1993;212(2):297–303. dellaconservazione Grasse. 104. Saranraj P, Stella D and Reetha D. 1977;54:381–388. Microbial cellulases and its 98. Ranalli A, Pollastri L, Contento S, applications – A review. International J Lucera L and Del Re P. Enhancing the of Biochem and Biotech Sci. 2012;1:1- quality of virgin olive oil by use of a 12.

437