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Physico-chemical properties, kinetic parameters, and glucose inhibition of several beta-glucosidases for industrial applications

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimization of biocatalysts and bioprocesses
Diandra de Andrades,1 Natália G. Graebin,1 Marco A. Z. Ayub,1 Roberto Fernandez- Lafuente,2 Rafael C. Rodrigues1
1Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil;
2Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049,10 Madrid, Spain

The β-glucosidase activities of nine commercial enzymatic preparations were compared by evaluating the effects of
pH, thermal stability, kinetic parameters, and glucose tolerance. The pH value of the reaction medium strongly
influenced the activities, stabilities, and glucose inhibition of the assayed β-glucosidases. Celluclast® 1.5 L showed the
highest activity on pNPG hydrolysis (70.55 U.mL-1 and 0.69, whereas Lallzyme Beta produced the highest
yields on cellobiose hydrolysis (93.4 %). Regarding the glucose tolerance and the pH, Pectinex Ultra SP-L presented
the maximum glucose tolerance at pH 3.0, keeping 54 % of its initial activity in the presence of 167 mmol.L-1 of
glucose, whereas at pH 7.0, Pectinex Smash XXL presented 44 % of activity at 222 mmol.L-1 of glucose. Pectinex
Smash XXL showed the highest KI (19.75 mol.L-1), whereas Celluclast® 1.5 L presented the highest KI/KM (19). Our
results suggest that these commercial preparations are promising sources of β-glucosidases, being Pectinex Smash
XXL indicated for applications at neutral pH because of its glucose tolerance and thermal stability at this pH value,
whereas Celluclast® 1.5 L, Lallzyme Beta, and Pectinex Ultra SP-L are more recommended for applications at acid

pH, such as in fruit juices and wine productions.
Keywords:β-glucosidase; glucose tolerance; kinetic parameters; enzyme activity.
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