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https://hdl.handle.net/2440/70420
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Type: | Journal article |
Title: | β-Glucoside metabolism in Oenococcus oeni: cloning and characterization of the phospho-β-glucosidase CelD |
Other Titles: | beta-Glucoside metabolism in Oenococcus oeni: Cloning and characterization of the phospho-beta-glucosidase CelD |
Author: | Capaldo, A. Walker, M. Ford, C. Jiranek, V. |
Citation: | Journal of Molecular Catalysis B: Enzymatic, 2011; 69(1-2):27-34 |
Publisher: | Elsevier Science BV |
Issue Date: | 2011 |
ISSN: | 1381-1177 1873-3158 |
Statement of Responsibility: | A. Capaldo, M. E. Walker, C. M. Ford and V. Jiranek |
Abstract: | In previous work, we reported characterization of a phospho-β- glucosidase gene bglD in a β-glucoside metabolizing operon in the oenologically important lactic acid bacterium Oenococcus oeni. Here we report a second phospho-β-glucosidase gene celD which has been cloned and expressed in Escherichia coli. This gene is found in a putative operon 6043 bp long encoding six genes designated celA to celF. Comparative sequence analyses of lactic acid bacteria suggest that the open reading frames of celA, B and F from the sequenced O. oeni PSU-1 encode phosphoenolpyruvate dependent phosphotransferase system (PEP-PTS) components IIB, IIA and IIC, respectively, which regulate the uptake and phosphorylation of β-glucosides across the cytoplasmic membrane. celE is speculated to have a regulatory function. celD was cloned and expressed in E. coli followed by purification of the gene product. The purified protein His-tagged CelD (485 residues, Mw = 55.8 kDa) has high homology to known phospho-β-glucosidases and has high activity towards the phosphorylated β-glucoside para-nitrophenol-β-d-glucopyranoside-6- phophate (pNPβG6P). CelD has an optimum pH between 4.0 and 5.0 and is most active at 40 °C. The gene celC was cloned, heterologously expressed and purified (481 residues, Mw = 55.7 kDa) but showed no significant activity towards pNPβG6P despite high sequence homology to celD and characterized phospho-β-glucosidases. Neither CelC nor CelD are active against non-phosphorylated β-glucosides. © 2010 Elsevier B.V. |
Keywords: | Oenococcus oeni Lactic acid bacteria Wine PEP-PTS Malolactic fermentation |
Rights: | © 2010 Elsevier B.V. All rights reserved. |
DOI: | 10.1016/j.molcatb.2010.12.006 |
Published version: | http://dx.doi.org/10.1016/j.molcatb.2010.12.006 |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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