The filamentous fungus Stachybotrys sp has been shown to possess a rich β-glucosidase system composed of five β-glucosidases. One of them was already purified to homogeneity and characterized. In this work, a second β-glucosidase was purified and characterized. The filamentous fungal A19 strain was fed-batch cultivated on cellulose, and its extracellular cellulases (mainly β-glucosidases) were analyzed. The purified enzyme is a monomeric protein of 78 kDa molecular weight and exhibits optimal activity at pH 6.0 and at 50°C. The kinetic parameters, K m and V max, on para-nitro-phenyl-β-d-glucopyranosid (p-NPG) as a substrate were, respectively, 1.846 ± 0.11 mM and 211 ± 0.08 μmol min−1 ml−1. One interesting feature of this enzyme is its high stability in a wide range of pH from 4 to 10. Besides its aryl β-glucosidase activity towards salicin, methylumbellypheryl-β-d-glucoside (MU-Glc), and p-NPG, it showed a true β-glucosidase activity because it splits cellobiose into two glucose monomers. This enzyme has the capacity to synthesize short oligosaccharides from cellobiose as the substrate concentration reaches 30% with a recovery of 40%. We give evidences for the involvement of a transglucosylation to synthesize cellotetraose by a sequential addition of glucose to cellotriose.
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This work is dedicated to the memory of our colleagues Said Hamdi and Nejib Trigui. We deeply thank Najla Masmoudi and Hédi Aouissaoui for their expert help in chromatographic analysis. Hafedh Belghith is thanked for helpful discussion and advises. Mosbeh Dardouri is thanked for his technical help.
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Saibi, W., Amouri, B. & Gargouri, A. Purification and biochemical characterization of a transglucosilating β-glucosidase of Stachybotrys strain. Appl Microbiol Biotechnol 77, 293–300 (2007). https://doi.org/10.1007/s00253-007-1141-3