β-Glucoside metabolism in Oenococcus oeni: cloning and characterisation of the phospho-β-glucosidase bglD
β-Glucoside metabolism in Oenococcus oeni: cloning and characterisation of the phospho-β-glucosidase bglD
The structural gene for a phospho-β-glucosidase from the oenologically important lactic acid bacterium (LAB) Oenococcus oeni has been cloned and its protein product characterised. This gene is found in a putative β-glucosidase operon of 2178 base pairs encoding 4 genes designated bglA to bglD. The bglA, B and C genes were not cloned and characterised, however, are thought to be phosphoenolpyruvate dependent phospho transferase system (PEP-PTS) components IIC, IIA and IIB which regulate the uptake, phosphorylation and translocation of β-glucosides across the cytoplasmic membrane. The cloned bglD was sequenced and expressed in Escherichia coli followed by purification. The purified bglD protein has 480 residues, a molecular mass of 55.5. kDa and shows high homology to known phospho-β-glucosidases. bglD exhibited high activity towards the phosphorylated β-glucoside para-nitrophenol-β-d-glucopyranoside-6-phosphate with a pH optimum of 5.5 and maintained similar levels of activity between temperatures of 4 °C and 40 °C. The enzyme was not active against non-phosphorylated β-glucosides.
Oenococcus oeni, Phospho-β-glucosidase, Phosphoenolpyruvate-phospho transferase system
476-482
Capaldo, A.
0567984a-c180-4d63-b06d-2d32c9146635
Walker, M.E.
bd0ff6e8-f7a5-45f4-84db-c90ea1a3e9ec
Ford, C.M.
af6fc87a-ef21-403b-a699-6eeb859ce65f
Jiranek, V.
8e5a8dfd-f5b2-43e3-928b-11dff324abc7
15 March 2011
Capaldo, A.
0567984a-c180-4d63-b06d-2d32c9146635
Walker, M.E.
bd0ff6e8-f7a5-45f4-84db-c90ea1a3e9ec
Ford, C.M.
af6fc87a-ef21-403b-a699-6eeb859ce65f
Jiranek, V.
8e5a8dfd-f5b2-43e3-928b-11dff324abc7
Capaldo, A., Walker, M.E., Ford, C.M. and Jiranek, V.
(2011)
β-Glucoside metabolism in Oenococcus oeni: cloning and characterisation of the phospho-β-glucosidase bglD.
Food Chemistry, 125 (2), .
(doi:10.1016/j.foodchem.2010.09.036).
Abstract
The structural gene for a phospho-β-glucosidase from the oenologically important lactic acid bacterium (LAB) Oenococcus oeni has been cloned and its protein product characterised. This gene is found in a putative β-glucosidase operon of 2178 base pairs encoding 4 genes designated bglA to bglD. The bglA, B and C genes were not cloned and characterised, however, are thought to be phosphoenolpyruvate dependent phospho transferase system (PEP-PTS) components IIC, IIA and IIB which regulate the uptake, phosphorylation and translocation of β-glucosides across the cytoplasmic membrane. The cloned bglD was sequenced and expressed in Escherichia coli followed by purification. The purified bglD protein has 480 residues, a molecular mass of 55.5. kDa and shows high homology to known phospho-β-glucosidases. bglD exhibited high activity towards the phosphorylated β-glucoside para-nitrophenol-β-d-glucopyranoside-6-phosphate with a pH optimum of 5.5 and maintained similar levels of activity between temperatures of 4 °C and 40 °C. The enzyme was not active against non-phosphorylated β-glucosides.
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More information
Accepted/In Press date: 8 September 2010
e-pub ahead of print date: 17 September 2010
Published date: 15 March 2011
Additional Information:
Funding Information:
This project was supported by Australia’s grapegrowers and winemakers through their investment body the Grape and Wine Research and Development Corporation , with matching funds from the Australian Government (projects UA 01/04 and UA 05/01). Thanks to Dennis Taylor and Peter Valente from the University of Adelaide for synthesising phosphorylated β-glucoside pNPβG6P.
Keywords:
Oenococcus oeni, Phospho-β-glucosidase, Phosphoenolpyruvate-phospho transferase system
Identifiers
Local EPrints ID: 482587
URI: http://eprints.soton.ac.uk/id/eprint/482587
ISSN: 0308-8146
PURE UUID: 41657cf0-b4e5-4d66-9bc5-32c768e4cfe1
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Date deposited: 10 Oct 2023 17:00
Last modified: 18 Mar 2024 04:12
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Contributors
Author:
A. Capaldo
Author:
M.E. Walker
Author:
C.M. Ford
Author:
V. Jiranek
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