CELLULASES

Cellulases

The EC number for this group of enzymes is EC 3.2.1.4. Class of enzymes that catalyze cellulolysis (i.e. the hydrolysis of cellulose). produced chiefly by fungi, bacteria, and protozoans few other types of organisms, such as some termites and the microbial intestinal symbionts of other termites

Hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, producing primary products glucose,cellobiose and cello oligosaccharides

Types and Action

Five types based on type of reaction catalysed Endocellulases : breaks internal bonds to disrupt the crystalline structure of cellulose and expose individual cellulose polysaccharide chains Exocellulases : cleaves 2 to 4 units from the ends of the exposed chains produced by endocellulase, resulting in the tetrasaccharides or disaccharides, such as cellobiose. There are two main types of exocellulases [or cellobiohydrolases (CBH)] CBHI works processively from the reducing end CBHII works processively from the nonreducing end of cellulose.

Cellobiase or beta-glucosidase: hydrolyses the exocellulase product into individual monosaccharides. Oxidative cellulases: depolymerize cellulose by radical reactions, as for instance cellobiose dehydrogenase (acceptor).

Cellulose phosphorylases depolymerize cellulose using phosphates instead of water.

Mechanism of cellulysis

Microorganisms producing cellulose

Celluloytic microbes are primarily carbohydrate degraders Cellulolytic microbes include bacteria Cellulomonas and Cytophaga , Cellovibrio Fungi: tricoderma reesei , humicola, penicillium, Aspergillus yield high level of cellulases.

microbes commercially exploited for cellulase preparations are T.reesei, H. insolens, A. niger , Thermomonospora fusca, Bacillus sp. Etc.

Cellulase Production

industrial demand for cellulases is being met by production methods using submerged fermentation (SmF) processes The cost of production in SmF systems is high and its uneconomical Alternative methods like SSF systems make use of aerobic microorganisms Production cost in the crude fermentation by SmF was about $ 20/kg by SSF it was only $ 0.2/kg if in situ fermentation was used

Carbon sources for commercial cellulase fermentation: -cellulosic biomass -straw -spent hulls of cereals and pulses - Rice or wheat bran - paper industry waste - various lignocellulosic residues Majority of processes are batch processes, attempts for fed batch and continuous process

Applications of Cellulase

Textile Industry Bio- Stonewashing of Denim garments - break down fabric (which is cellulose) - releases dye Digesting off small fiber ends - better finishing Providing localized variation in colour density. H. insolens cellulase is most commonly imployed

Laundry and Detergents Cellulase: EGIII and CBH I commonly used in detergents Cellulase preparation from Humicola : active under mild alkaline condition and elevated temperatures are added in washing powder and in detergents.

Food and Animal Feed - Extraction and clarification of food and vegetables juices - Production of fruit nectars and purees - Extraction of olive oil - Glucanase: improves malting of barley in beer and wine production - Used in carotenoid extraction for production of food coloring agent - Cellulase with hemicellulase and pectinase is used to improve nutritive quality of forages - Increases digestibility of animal feed

Pulp and Paper industry - used for Biomechanical pulping - de-inking of recycled fibers - improving drainage and runnability of paper mills - used for characterization of pulp fibers - preparation of easily biodegradable cardboards.

Production of biofuel (U tilization of lignocellulosic waste) S teps: 1) Pretreatment of lignocellulosic residue to remove lignin and hemicellulase fraction 2) Cellulase treatment at 50C to hydrolyse cellulosic residues to generate fermentable sugars 3) Use of fermentative micro-organism to produce alcohol from the hydrolysed cellulosic material

O ther applications -employed in formulations for removal of industrial slime - in research for generation of protoplast - for generation of antibacterial chitooligosaccharides (used in food preservation)

Challenges in cellulase research

Cellulase production is not cost effective less efficient in conversion of natural lignocellulosic material to essential metabolites Current commercial preparation of cellulose is economically not feasible

Major goals for future cellulase research Reduction in cost of cellulase production - optimizing growth conditions - lactose as an inducer of cellulase genes -use of mixed culture enhances production Genetic improvement of cellulase performance -Using cbh1 promoter of T.resei for over expression of cellulases

References

R. K. Sukumaran, Reeta Rani Singhania and Ashok Pandey (2005). Microbial cellulases Production, applications and challenges. Journals of Scientific & Industrial Research 64: 832-844 K. Reczey, Zs. Szengyel, R. Eklund & G. Zacchi (1996). Cellulase Production by T. Reesei. Bioresource Technology 57: 25-30 Bhat M K & Bhat S (1997). Cellulose degrading enzymes and their potential industrial applications. Bioethanol Adv 15: 583-620

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