Substrate conditions that influence the assays used for determining the ?-glucosidase activity of cellulolytic microorganisms

Culture filtrates from Trichoderma harzianum E58, T. reesei CL 847 and Penicillium sp. C 462 were assayed for beta-glucosidase activity using a range of substrates and sugar analysis methods. Although sugar analyses by the dinitrosalicylic acid (DNS) and Nelson-Somogyi methods gave a similar profile, when increasing concentrations of salicin were assayed, considerably higher values were obtained with the DNS assay. The salicin concentration used for the assay greatly influenced the final beta-glucosidase values with higher values obtained for T. harzianum E58 and T. reesei CL 847 at substrate concentrations of 1 mg/mL while optimum values for Penicillium sp. C 462 were obtained at substrate concentrations greater than 3 mg/mL. Low concentrations of salicin and p-nitro-phenyl-beta-D-glucopyranoside (PNPG) gave the same response as cellobiose. Cellobiose should be used at concentrations greater than 3.74 mg/mL to avoid substrate limitation of the beta-glucosidase assay.