Lactobacillus plantarum WCFS1 β-fructosidase : evidence for an open funnel-like channel through the catalytic domain with importance for the substrate selectivity
Articolo
Data di Pubblicazione:
2016
Citazione:
Lactobacillus plantarum WCFS1 β-fructosidase : evidence for an open funnel-like channel through the catalytic domain with importance for the substrate selectivity / E.O. Mendoza Llerenas, D.J. Pérez, Z. Gómez Sandoval, P. Escalante Minakata, V. Ibarra Junquera, R.S. Razo Hernández, V. Capozzi, P. Russo, G. Spano, D. Fiocco, J.A. Osuna Castro, A. Moreno. - In: APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY. - ISSN 0273-2289. - 180:6(2016 Nov), pp. 1056-1075. [10.1007/s12010-016-2152-2]
Abstract:
β-Fructosidase, a glycoside hydrolase of a biotechnologically important strain, was studied for its biochemical, physicochemical, and three-dimensional structure characteristics. This enzyme was heterologously expressed in Escherichia coli as a C-terminal His-tagged protein (SacB). β-Fructosidase catalyzes the cleavage of glycoside bonds toward certain carbohydrates with β-fructofuranosyl linkages; however, SacB exhibited selectivity toward sucrose and an optimum activity at pH 6.0–6.5 and 37 °C. In such optimum enzymatic activity conditions, the SacB was commonly observed as a monodisperse protein by dynamic light scattering (DLS). As β-fructosidase belongs to glycoside hydrolase family 32 (GH32), a β-sandwich and a five-bladed β-propeller domain are typical predicted folds in its structure. Docking and molecular dynamic simulations revealed for the first time a funnel-like channel perfectly exposed in the β-propeller domain of the Lactobacillus plantarum β-fructosidase (this allows the interaction between its entire catalytic triad and substrates that are larger than sucrose). In contrast, SacB showed a closed central tunnel collaterally induced by its His-tag.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
computer simulations; invertase; lactobacillus plantarum; selectivity; β-fructosidase; β-propeller domain; biotechnology; bioengineering; biochemistry; applied microbiology and biotechnology; molecular biology
Elenco autori:
E.O. Mendoza Llerenas, D.J. Pérez, Z. Gómez Sandoval, P. Escalante Minakata, V. Ibarra Junquera, R.S. Razo Hernández, V. Capozzi, P. Russo, G. Spano, D. Fiocco, J.A. Osuna Castro, A. Moreno
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