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Basic characterization and partial purification of β-glucosidase from cell-free extracts of Oenococcus oeni ST81.
Lett Appl Microbiol 2012; 55(3):247-55LA

Abstract

AIMS

This study was designed to characterize a β-glucosidase of Oenococcus oeni ST81, a strain isolated from a Spanish wine of the origin appellation Ribeira Sacra.

METHODS AND RESULTS

The β-glucosidase of O. oeni ST81 seems to have a periplasmic localization into the cells. This activity was strongly inhibited by gluconic acid, partially inhibited by glucose and not inhibited by fructose, lactate, malate, mannitol or sorbitol. Ethanol increased the activity of this enzyme up to 147%. Among the several metal ions assayed, only Fe²⁺ (10 mmol l⁻¹) and Cu²⁺ (5 mmol l⁻¹) exhibited a partial inhibitory effect (40%). This enzyme was partially purified using a combination of ammonium sulfate precipitation and chromatographic methods. The single peak because of β-glucosidase in all chromatographic columns indicates the presence of a single enzyme with an estimated molecular mass of 140 kDa. The calculated K(m) and V(max) values for 4-nitrophenyl-β-D-glucopyranoside were 0·38 mmol l⁻¹ and 5·21 nmol min⁻¹, respectively. The enzyme was stable at pH 5·0 with a value of t(1/2) = 50 days for the crude extract.

CONCLUSIONS

The β-glucosidase of O. oeni ST81 is substantially different from those characterized from other wine-related lactic acid bacteria (LAB), such as Lactobacillus plantarum and Lactobacillus brevis; however, it appears to be closely related to a β-glucosidase from O. oeni ATCC BAA-1163 cloned into Escherichia coli. The periplasmic localization of the enzyme together with its high tolerance to ethanol and fructose, the low inhibitory effect of some wine-related compounds on the enzymatic activity and long-term stability of the enzyme could be of interest for winemaking.

SIGNIFICANCE AND IMPACT OF THE STUDY

Information regarding a β-glucosidase from O. oeni ST81 is presented. Although the release of aroma compounds by LAB has been demonstrated, little information exists concerning the responsible enzymes. To our knowledge, this study contains the first characterization of a native β-glucosidase purified from crude extracts of O. oeni ST81.

Authors+Show Affiliations

Departamento de Química Analítica, Nutrición y Bromatología-Tecnología de Alimentos, Escuela Politécnica Superior, Universidad de Santiago de Compostela, Lugo, Spain. juan.mesas@usc.esNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

22748149

Citation

Mesas, J M., et al. "Basic Characterization and Partial Purification of Β-glucosidase From Cell-free Extracts of Oenococcus Oeni ST81." Letters in Applied Microbiology, vol. 55, no. 3, 2012, pp. 247-55.
Mesas JM, Rodríguez MC, Alegre MT. Basic characterization and partial purification of β-glucosidase from cell-free extracts of Oenococcus oeni ST81. Lett Appl Microbiol. 2012;55(3):247-55.
Mesas, J. M., Rodríguez, M. C., & Alegre, M. T. (2012). Basic characterization and partial purification of β-glucosidase from cell-free extracts of Oenococcus oeni ST81. Letters in Applied Microbiology, 55(3), pp. 247-55. doi:10.1111/j.1472-765X.2012.03285.x.
Mesas JM, Rodríguez MC, Alegre MT. Basic Characterization and Partial Purification of Β-glucosidase From Cell-free Extracts of Oenococcus Oeni ST81. Lett Appl Microbiol. 2012;55(3):247-55. PubMed PMID: 22748149.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Basic characterization and partial purification of β-glucosidase from cell-free extracts of Oenococcus oeni ST81. AU - Mesas,J M, AU - Rodríguez,M C, AU - Alegre,M T, Y1 - 2012/07/24/ PY - 2012/7/4/entrez PY - 2012/7/4/pubmed PY - 2013/6/1/medline SP - 247 EP - 55 JF - Letters in applied microbiology JO - Lett. Appl. Microbiol. VL - 55 IS - 3 N2 - AIMS: This study was designed to characterize a β-glucosidase of Oenococcus oeni ST81, a strain isolated from a Spanish wine of the origin appellation Ribeira Sacra. METHODS AND RESULTS: The β-glucosidase of O. oeni ST81 seems to have a periplasmic localization into the cells. This activity was strongly inhibited by gluconic acid, partially inhibited by glucose and not inhibited by fructose, lactate, malate, mannitol or sorbitol. Ethanol increased the activity of this enzyme up to 147%. Among the several metal ions assayed, only Fe²⁺ (10 mmol l⁻¹) and Cu²⁺ (5 mmol l⁻¹) exhibited a partial inhibitory effect (40%). This enzyme was partially purified using a combination of ammonium sulfate precipitation and chromatographic methods. The single peak because of β-glucosidase in all chromatographic columns indicates the presence of a single enzyme with an estimated molecular mass of 140 kDa. The calculated K(m) and V(max) values for 4-nitrophenyl-β-D-glucopyranoside were 0·38 mmol l⁻¹ and 5·21 nmol min⁻¹, respectively. The enzyme was stable at pH 5·0 with a value of t(1/2) = 50 days for the crude extract. CONCLUSIONS: The β-glucosidase of O. oeni ST81 is substantially different from those characterized from other wine-related lactic acid bacteria (LAB), such as Lactobacillus plantarum and Lactobacillus brevis; however, it appears to be closely related to a β-glucosidase from O. oeni ATCC BAA-1163 cloned into Escherichia coli. The periplasmic localization of the enzyme together with its high tolerance to ethanol and fructose, the low inhibitory effect of some wine-related compounds on the enzymatic activity and long-term stability of the enzyme could be of interest for winemaking. SIGNIFICANCE AND IMPACT OF THE STUDY: Information regarding a β-glucosidase from O. oeni ST81 is presented. Although the release of aroma compounds by LAB has been demonstrated, little information exists concerning the responsible enzymes. To our knowledge, this study contains the first characterization of a native β-glucosidase purified from crude extracts of O. oeni ST81. SN - 1472-765X UR - https://www.unboundmedicine.com/medline/citation/22748149/Basic_characterization_and_partial_purification_of_β_glucosidase_from_cell_free_extracts_of_Oenococcus_oeni_ST81_ L2 - https://doi.org/10.1111/j.1472-765X.2012.03285.x DB - PRIME DP - Unbound Medicine ER -