Activation of acetate in acetate-grown Methanosarcina thermophila: purificationm and characterization of acetate kinase

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dc.contributor.authorAceti, David Johnen
dc.contributor.committeechairFerry, James G.en
dc.contributor.committeememberWilkins, Tracy D.en
dc.contributor.committeememberDean, Dennis R.en
dc.contributor.committeememberWhite, Robert H.en
dc.contributor.departmentAnaerobic Microbiologyen
dc.date.accessioned2014-03-14T21:28:49Zen
dc.date.adate2013-02-07en
dc.date.available2014-03-14T21:28:49Zen
dc.date.issued1980-04-05en
dc.date.rdate2013-02-07en
dc.date.sdate2013-02-07en
dc.description.abstractExtracts of acetate-grown <u>Methanosarcina thermoghila</u> were assayed for the presence of enzymes which might catalyze a proposed activation of acetate as the initial step in the pathway of methanogenesis from acetate by that organism. Acetate kinase and phosphate acetyltransferase activities of 4.9 and 49 μmoles of product/min/mg protein, respectively, were detected. Acetate kinase was purified 102- fold to a specific activity of 656 μmoles ADP formed/min/mg protein and was essentially homogeneous by denaturing gel electrophoresis. The native enzyme (Mr 94,000) was an α₂ homodimer with a subunit Mr of 53,000. Activity was optimal between pH 7.0 and 7.4 and was stable to heating at 70°C for 15 min. The apparent Km for acetate was 22 mM (Vmax = 668 μmoles ADP/min/mg protein) and 2.8 mM for ATP (Vmax = 777 pmoles ADP/min/ protein). The enzyme phosphorylated propionate at 602 of the rate with acetate but was unable to use formate. TTP, ITP, UTP, GTP, and CTP replaced ATP as the phosphoryl donor to acetate. One of several divalent cations was required for activity; the maximum rate was obtained with Mn2+.en
dc.description.degreeMaster of Scienceen
dc.format.extentviii, 79 leavesen
dc.format.mediumBTDen
dc.format.mimetypeapplication/pdfen
dc.identifier.otheretd-02072013-040125en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-02072013-040125/en
dc.identifier.urihttp://hdl.handle.net/10919/40983en
dc.language.isoenen
dc.publisherVirginia Techen
dc.relation.haspartLD5655.V855_1989.A254.pdfen
dc.relation.isformatofOCLC# 20060396en
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subject.lccLD5655.V855 1989.A254en
dc.subject.lcshMethylotrophic bacteriaen
dc.subject.lcshMethylotrophic microorganismsen
dc.titleActivation of acetate in acetate-grown Methanosarcina thermophila: purificationm and characterization of acetate kinaseen
dc.typeThesisen
dc.type.dcmitypeTexten
thesis.degree.disciplineAnaerobic Microbiologyen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

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