Cellulase isolation methods

This study describes an absolute method for the evaluation of endo-/ -glucanase (Cx) or endocellulase activities in the cellulase complex without the need of a time-consuming isolation of the endocellulase fractions. In the method proposed by Almin et al. (3), an assumption was needed in the theoretical derivation of endocellulase activities, namely, that Mv/Mn is constant in the initial stages of the enzymic reactions. We found, however, a linear relationship between Mv and STn. They used CMC as substrate, and since the mode of action of endocellu-lases on CMC can be regarded as similar to that on HEC, it is possible that their assumption is not fulfilled. 

Selective Screening Methods for the Isolation of High Yielding Cellulase Mutants of Trichoderma reesei... 

T ine structural studies on woody cell walls attacked by ectoenzymes of fungi in situ are numerous (cf. 1,2). In contrast, investigations on the selective degradation of cell walls by enzymes isolated from fungi are few. Jutte and Wardrop (3) attempted the use of crude commercial cellu-lase preparations to determine the degradation pattern of Valonia cellulose and beechwood fibers. Similar use of commercial preparations of enzymes was made by Reis and Roland (4) to evaluate the nature of diverse cell walls and to show the distribution of polysaccharides. An endo-/ -l,4-xylanase with specific xylanolytic activities was isolated from a commercial cellulase preparation using chromatographic methods and... 

In earlier methods, sodium carbonate or sodium hydroxide solutions and a low shear rate or water and a high shear rate were used.6 The alkali isolation procedure is shown in Figure 15.1a. A modification involving use of enzymes (protease and/or cellulase) (Figure 15.1b) gave a starch yield of 86%.6 With another method employing a protease, 92-98% of the starch was recovered.11... 

For the last few years, we have investigated the cellulase components of Pseudomonas fluorescens var. cellulosa in special regard to the physiological relation of their synthesis and localization to the cultural conditions. This pseudomonad is an aerobic mesophilic cellulose-decomposing bacterium isolated earlier by Ueda et al. (39) from field soil. Some of the enzymatic properties of cellulases obtained from it have previously been reported (28). In the present review, the results of our recent studies (42) are described and discussed together with the related works of other authors. [In these studies, activities of cellulase and aryl / -glucosidase were assayed by the same methods as described in a recent paper (31), and that of amylase by the blue value method modified by Fuwa (10)]. 

Cell walls can be rapidly isolated from plant tissues by a modification of the method of Van Soest and Wine which involves extraction with neutral detergent. Most investigators have treated isolated cell walls with alkali to release phenolic acids as their sodium salts. The acids can also be released as their carbohydrate esters by degrading the walls with a commercial cellulase preparation from fungal sources (Basidiomycete, Oxyporous spp.) having... 

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