The enzyme was completely inhibited by iron chloride, silver nitrate and SDS in both concentrations tested. In the assay conditions, the denaturing action of SDS probably affected the integrity of the enzyme tridimensional structure which is fundamental for its catalytic activity. Inhibition caused by SDS (1 and 10 mM) was also demonstrated by Li, Jiang, Fan, and Liu (2012) for cloned β-glucosidase using metagenomic DNA from mangrove Torin 1 manufacturer soil. D. hansenii UFV-1 β-glucosidase activity was greatly increased by β-mercaptoethanol,

glucose, urea and aluminium chlorid at both concentrations tested. Non-inhibition by EDTA implies that divalent cations are not essential to enzyme activity ( Chen, Li, & Zong, 2012b) and it is not a metalloenzyme. Many works reported that EDTA does not inhibit β-glucosidases as in the case of Pyrococcus furiosus β-glucosidase that was considered metal-independent ( Yeom et al., 2012). β-Mercaptoethanol was the agent which best promoted enzyme activation in both final concentrations tested. The activation by this Tenofovir datasheet reducing agent can be explained

by the fact that some reduced chemical ligations in the enzyme structure are favourable for the catalytic activity. Calcium and magnesium have a stimulatory effect on D. hansenii UFV-1 β-glucosidase. It has been reported that these two ions are enhancers of β-glucosidase activity ( Oyekola, Ngesi, & Whiteley, 2007). Glucose was found to be a competitive inhibitor of D. hansenii UFV-1 β-glucosidase and the Ki value was 11.36 mM. In general, β-glucosidases are inhibited

by glucose and this inhibition is competitive ( Yang et al., 2004). Soy molasses is a by-product generated in the production of soy protein concentrate, in which isoflavones and other phytochemicals are enriched all (Hosny & Rosazza, 1999). This by-product in the soy industry is used as an inexpensive animal feed, but the processing and use of soy molasses as a functional food has been suggested (Najafpour & Shan, 2003). The potential of D. hansenii UFV-1 intracellular β-glucosidase to hydrolyze isoflavones in soy molasses to their aglycon forms was demonstrated for the free β-glucosidase and the alginate immobilised cells containing this enzyme ( Table 4). Prior to hydrolysis, glucoside isoflavones were predominant in the soy molasses, representing approximately 80%, where aglycones made up about 10%. After 2 h of treatment with the free or immobilised β-glucosidase the isoflavone glucosides were almost completely hydrolyzed after which there remained about 3% of these compounds. There was no change in the amounts of isoflavone glucosides and aglycones after 4 or 8 h of enzymatic treatment of soy molasses compared to the assay after 2 h of hydrolysis (data not shown). This indicates that a short incubation period is preferred over a prolonged incubation with the free or immobilised enzyme.