β-Glucoside metabolism in Oenococcus oeni: cloning and characterization of the phospho-β-glucosidase CelD

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dc.contributor.authorCapaldo, A.
dc.contributor.authorWalker, M.
dc.contributor.authorFord, C.
dc.contributor.authorJiranek, V.
dc.date.issued2011
dc.description.abstractIn 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.
dc.description.statementofresponsibilityA. Capaldo, M. E. Walker, C. M. Ford and V. Jiranek
dc.identifier.citationJournal of Molecular Catalysis B: Enzymatic, 2011; 69(1-2):27-34
dc.identifier.doi10.1016/j.molcatb.2010.12.006
dc.identifier.issn1381-1177
dc.identifier.issn1873-3158
dc.identifier.orcidWalker, M. [0000-0002-6934-3787]
dc.identifier.orcidFord, C. [0000-0003-1617-2977]
dc.identifier.orcidJiranek, V. [0000-0002-9775-8963]
dc.identifier.urihttp://hdl.handle.net/2440/70420
dc.language.isoen
dc.publisherElsevier Science BV
dc.rights© 2010 Elsevier B.V. All rights reserved.
dc.source.urihttps://doi.org/10.1016/j.molcatb.2010.12.006
dc.subjectOenococcus oeni
dc.subjectLactic acid bacteria
dc.subjectWine
dc.subjectPEP-PTS
dc.subjectMalolactic fermentation
dc.titleβ-Glucoside metabolism in Oenococcus oeni: cloning and characterization of the phospho-β-glucosidase CelD
dc.title.alternativebeta-Glucoside metabolism in Oenococcus oeni: Cloning and characterization of the phospho-beta-glucosidase CelD
dc.typeJournal article
pubs.publication-statusPublished

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