Location of the β-galactosidase of the yeast Kluyveromyces marxianus var. marxianus ATCC 10022

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dc.contributor.authorBacci, M. [UNESP]
dc.contributor.authorSiqueira, C. G. [UNESP]
dc.contributor.authorAntoniazi, S. A.
dc.contributor.authorUeta, J.
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.date.accessioned2014-05-27T11:18:08Z
dc.date.available2014-05-27T11:18:08Z
dc.date.issued1996-12-01
dc.description.abstractDuring the growth of Kluyveromyces marxianus var. marxianus ATCC 10022 on lactose, peaks of glucose, but not β-galactosidase activity, were detected in culture medium. Harvested and washed whole cells produced glucose and galactose from lactose, or ortho-nitro-phenol from the chromogenic substrate ortho-nitro-phenyl-β-D-galactopyranoside (ONPG), indicating that β-galactosidase is physically associated with cells. ONPG hydrolysis by whole cells presented a monophasic kinetics (Km 36.6 mM) in lactose exponential growth phase cells, but a biphasic kinetics (Km 0.2 and 36.6 mM) in stationary growth phase cells. Permeabilization with digitonin or disruption of cells from both growth phases led to monosite ONPG hydrolysis (Km 2.2 to 2.5 mM), indicating that β-galactosidase is not located in the periplasm. In addition, the energy inhibitors fluoride or arsenate, as well as the uncouplercarbonyl cyanide m-chlorophenylhydrazone (CCCP) prevented ONPG hydrolysis by whole cells. These findings indicate that energy coupled transmembrane transport is the rate-limiting step for intracellular ONPG cleavage. The taxonomic and physiologic implications of the exclusive intracellular location of β-galactosidase of K. marxianus var. marxianus ATCC 10022 are discussed. © 1996 Kluwer Academic Publishers.en
dc.description.affiliationDepartment of Biochemistry and Microbiology Biosciences Institute of Rio Claro UNESP, PO Box 199, 13506-900 Rio Claro, SP
dc.description.affiliationDepartment of Pharmaceutical Sciences Faculty of Pharmaceutical Sciences of Ribeirão Preto USP, Av. do Café, s/n, 14100-906, Ribeirão Preto, SP
dc.description.affiliationUnespDepartment of Biochemistry and Microbiology Biosciences Institute of Rio Claro UNESP, PO Box 199, 13506-900 Rio Claro, SP
dc.format.extent357-361
dc.identifierhttp://dx.doi.org/10.1007/BF00399624
dc.identifier.citationAntonie van Leeuwenhoek, International Journal of General and Molecular Microbiology, v. 69, n. 1, p. 357-361, 1996.
dc.identifier.doi10.1007/BF00399624
dc.identifier.issn0003-6072
dc.identifier.scopus2-s2.0-0040349559
dc.identifier.urihttp://hdl.handle.net/11449/64888
dc.language.isoeng
dc.relation.ispartofAntonie Van Leeuwenhoek International Journal of General and Molecular Microbiology
dc.relation.ispartofjcr1.588
dc.relation.ispartofsjr0,834
dc.rights.accessRightsAcesso restrito
dc.sourceScopus
dc.subjectβ-galactosidase
dc.subjectKluyveromyces marxianus
dc.subjectLactose
dc.subjectONPG
dc.subjectTransport
dc.subjectYeast
dc.titleLocation of the β-galactosidase of the yeast Kluyveromyces marxianus var. marxianus ATCC 10022en
dc.typeArtigo
dcterms.licensehttp://www.springer.com/open+access/authors+rights
unesp.author.orcid0000-0002-5619-1411[1]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Rio Claropt

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