Systematic studies of the interactions between a model polyphenol compound and microbial beta-glucosidases

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2017
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da Silva, Viviam M.
Sato, Juliana A. P.
Araujo, Juscemacia N.
Squina, Fabio M.
Muniz, Joao R. C.
Riske, Karin A. [UNIFESP]
Garcia, Wanius
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Lignin is a major obstacle for cost-effective conversion of cellulose into fermentable sugars. Non-productive adsorption onto insoluble lignin fragments and interactions with soluble phenols are important inhibition mechanisms of cellulases, including beta-glucosidases. Here, we examined the inhibitory effect of tannic acid (TAN), a model polyphenolic compound, on beta-glucosidases from the bacterium Thermotoga petrophila (TpBGL1 and TpBGL3) and archaeon Pyrococcus furiosus (PfBGL1). The results revealed that the inhibition effects on beta-glucosidases were TAN concentration-dependent. TpBGL1 and TpBGL3 were more tolerant to the presence of TAN when compared with PfBGL1, while TpBGL1 was less inhibited when compared with TpBGL3. In an attempt to better understand the inhibitory effect, the interaction between TAN and beta-glucosidases were analyzed by isothermal titration calorimetry (ITC). Furthermore, the exposed hydrophobic surface areas in beta-glucosidases were analyzed using a fluorescent probe and compared with the results of inhibition and ITC. The binding constants determined by ITC for the interactions between TAN and beta-glucosidases presented the same order of magnitude. However, the number of binding sites and exposed hydrophobic surface areas varied for the beta-glucosidases studied. The binding between TAN and beta-glucosidases were driven by enthalpic effects and with an unfavorable negative change in entropy upon binding. Furthermore, the data suggest that there is a high correlation between exposed hydrophobic surface areas and the number of binding sites on the inhibition of microbial beta-glucosidases by TAN. These studies can be useful for biotechnological applications.
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Plos One. San Francisco, v. 12, n. 7, p. -, 2017.
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