Cellulases structural features

Structural features met in some cellulases include an a,a barrel111 similar to that of glucoamylase (Fig. 2-29) and, in a cellobiohydrolase,101 a 5-nm-long tunnel into which the cellulose chains must enter. Ten well-defined subsites for glycosyl units are present in the tunnel.101 A feature associated with this tunnel is processive action, movement of the enzyme along the chain without dissociation,105 a phenomenon observed long ago for amylases (see Section 9) and often observed for enzymes acting on nucleic acids. 

The core-enzymes, prepared in our laboratory, and containing the active centers, were successfully crystallized (Dr. Jones, Uppsala, communicated) and tertiary structures will be described in the near future. Chemical modification studies on these enzymes are currently being undertaken in our laboratory identification of important catalytic residues and location of the active centers will lead to more functional information on these enzymes. Other cellulases such as some endoglucanases from Clostridium thermocel-lum (EG A, EG B, EG D) (10) and EngA and Exg from Cellulomonas fimi (19) also contain sequences of conserved, terminally located and sometimes reiterated, amino acids. Some of these sequences are preceded by proline-serine rich domains. Thus, a bistructural-bifunctional organization seems to be a rather common feature among cellulases, at least for EngA and Exg from C. fimi and the enzymes from Trichoderma reesei. 

The cellulase complex diffuses through the pore system to the microfibrils, attacks the cellulose chains and hydrolyses each chain to the end. The diflerences in the efficacy of cellulases on various fibres are dependent on number of factors such as the amounts of non-cellulosic wood pulp-derived matter, the degree ol polymerisation, the type and degree of crystallinity, and the type and number of chemical substitutions to the cellulose [27-30]. Key features for the cellulose substrate are crystallinity, accessible surface area and pore dimensions [31 ]. Variation of any of these factors, e.g., structural changes of cellulose substrate by pre-treatments, will influence the course of the entire degradation process [32, 33]. 

That such features of cellulose as the crystal lattice form are significant determinants of cellulase action has only recently been established (19), although a great deal remains to be learned about the enzymatic importance of cellulose fine structure. It is clearly established, however, that each of the three water-stable crystal forms of cellulose is distinct in the rate at which it is hydrolyzed and in its properties as an inducer of cellulase. For example the Trichoderma viride cellulase from culture extracts exhibits a lower activation energy in attacking the crystal lattice form used in culture growth, than in attacking the other lattice forms (Table I). 

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