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Activation and Cell-Surface Localization of Certain ß-Glucosidases of the Ruminal Flora

Departments of Biochemistry and Nutrition and of Biology, Virginia Polytechnic Institute, Blacksburg

ABSTRACT

Kinetic and inhibition studies of the cellulases in crude extracts of ruminal ingesta indicated the participation of sulfhydryl groups and polyvalent metal ions in enzymic hydrolysis of cellulose. As a result, the assay substrate has been modified by inclusion of ferric chloride. A minimum estimate was obtained of the proportion of the cellulolytic activity in ruminal ingesta which is intimately associated with bacterial cell surfaces. A series of fractionations employing washing of the bacterial cells by centrifugation, rupture in a French pressure cell, and lysis using desoxycholate yielded an estimate of 5%. Reasons for suspecting that surface-bound cellulases may occur in considerably larger proportions were discussed.

¹ With the technical assistance of Blanche Ching-yi Wu, Laboratoire National d'Hygiene Publique, Port au Prince, Haiti.