Cellulases capability

T. reesei, a saprophytic fungus, is capable of utilizing a variety of carbohydrates. Yet, only a few carbohydrates induce cellulase production. Inducers include cellulose, cellulose derivatives, lactose, and sophorose (31,32). Mandels and Reese (31,32) studied the inducibility of various sugars and found that sophorose is an excellent cellulase inducer in T. reesei while having little effect in other fungi or bacteria. On further examination they found that trace amounts of sophorose present in glucose caused the apparent ability of glucose to be a cellulase inducer in T. reesei. 

To hydrolyze crystalline cellulose efficiently by enzymatic means, the inaccessibility of crystalline structures must be overcome. T. reesei and some other true cellulolytic microorganisms produce a cellulase complex that is capable of efficiently hydrolyzing crystalline cellulose. One explanation of this capability was first proposed by Mandels and Reese (7). In this model, two factors, Ci and C worked together to disrupt and hydrolyze cellulose. Ci first disrupted the crystalline structure of the cellulose while Cx attacked the available sites formed by Ci. In other words, Ci and C exhibit synergism in hydrolyzing cellulose. Since then, the combined action of cellobiohydrolase ( Ci ) and endoglucanase ( C ) has been identified as the source of the apparent synergism (6,26,55). 

NG14. This mutant is capable of elaborating 15 FP units/mL with a productivity of 45 units/L/hr. The final concentration of soluble extracellular protein is 21.2 mg/mL. Samples of this enzyme preparation were sent to G. Pettersson at the University of Uppsala, Sweden, for quantification of each of the enzymes in the cellulase complex using purified antibodies to the individual enzymes. The quantitative antigen-antibody reaction showed that 600 mg/g of this enzyme preparation was one enzyme, cellobiohydrolase. This represents a yield of 13 g/L of cellobio-hydrolase, which is a 100-fold increase over the amount of cellobiohydrolase obtained with strain QM 9414 (130 mg/L) the best previously existing cellulase mutant (G. Pettersson, personal communication). 

Many fungi are capable of producing extracellular enzymes that can degrade cellulose. They are Trichoderma (T) reesei, T. viride, T. koningii, T. lignorum, Penicillium funiculosum, Fusarium solani, Sclerotium rolfsii, and so on. Bacterial species such as Cellulomonas along with Clostridium thermocellum can also produce cellulases (Marsden and Gray, 1986). 

Fungal cellulase enzyme systems capable of efficiently catalyzing the hydrolytic degradation of crystalline cellulose are typically composed of endo-acting cellulases (EGs), exo-acting cellulases (CBHs), and at least one cellobiase (1-6). The CBHs are typically the predominant enzymes, on a mole fraction basis, in such systems (7). Consequently, the CBHs have been the focus of many studies (8). The three-dimensional structure of prototypical CBHs is known (9-12) and their specificities are, in general, well characterized (13,14). However, mechanism-based kinetic analyses of CBH-catalyzed cellulose saccharification are rather limited (15,16). Studies of this latter type are particularly difficult owing to the inherent complexity of native cellulose substrates. 

Despite of their lack in hydrolytic activity, the CBDs of the cellulases CenA and Cex from C. fimi have been found to be capable of disrupting cotton fibers and releasing small particles from the substrate (Kilburn et al., 1993 Din et al., 1994a). This disruptive effect does not seem to be a general characteristic of CBM, since it was only found with CBD from cellulose from a Penicillium sp. (Gao et al., 2001). Anyhow, a synergism with the catalytic module and enhanced degradation capacity were reported (Din et al., 1994a). 

High concentrations of stabilizing salts such as NaCl and Na2S04 were reported to increase, and chaotropic salts to decrease, the affinity of bacterial and fungal cellulases to crystalline cellulose. Denaturing agents are capable of eluting cellulases from the cellulosic substrate (Otter et al., 1989). 

The original cellulases used in denim washing were the crude enzymes of Trichoderma and Humicola, referred to as acid and neutral cellulase, respectively, based on the optimum pH range of use of the enzymes, which was pH 4 to 5 for the acid cellulase and 6 to 7 for the neutral cellulase. The Trichoderma cellulase, comprising the more complete set of EG and CBH components capable of the full hydrolysis of cellulose, works more quickly and is capable of a greater degree of abrasion and fading of the blue dye color than the Humicola cellulase. The Trichoderma cellulase also achieves certain desired finishes (appearances) that the Humicola cellulase does not. 

The raw materials required to produce cellulase are cheap and abundant. Natural cellulosic materials such as plant stems and corn cobs can be used as the carbon source. The composition of the medium is simple and of low cost. Because of the high capability of cellulosic materials to buffer the pH value, it is not necessary to add additional expensive buffer solution. 

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