Purification and properties of glutamate synthase from Bacillus licheniformis

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dc.contributor.authorSchreier, Harold J.
dc.contributor.authorBernlohr, R. W .
dc.date.accessioned2019-03-15T19:14:30Z
dc.date.available2019-03-15T19:14:30Z
dc.date.issued1984-11
dc.description.abstractGlutamate synthase [L-glutamate:NADP+ oxidoreductase (transaminating); EC 1.4.1.13] (GItS) was purified to homogeneity from BaciUus licheniformis A5. The native enzyme had a molecular weight of approximately 220,000 and was composed of two nonidentical subunits (molecular weights, -158,000 and -54,000). The enzyme was found to contain 8.1 ± 1 iron atoms and 8.1 + 1 acid-labile sulfur atoms per 220,000-dalton dimer. Two flavin moieties were found per 220,000-dalton dimer, with a ratio of flavin adenine dinucleotide to flavin mononucleotide of 1.2. The UV-visible spectrum of the enzyme exhibited maxima at 263,380 and 450 nm. The GitS from B. Iicheniformis had a requirement for NADPH, oa-ketoglutarate, and glutamine. Classical hyperbolic kinetics were seen for NADPH affinity, which resulted in an apparent Km value of 13 ,uM. Nonhyperbolic kinetics were obtained for a-ketoglutarate and glutamine affinities, and the reciprocal plots obtained for these substrates were biphasic. The apparent Km values obtained for glutamine were 8 and 100 ,LM, and the apparent Km values obtained for a-ketoglutarate were 6 and 50 ,uM. GltS activity was found to be relatively insensitive to inhibition by amino acids, keto acids, or various nucleotides. L-Methionine-DLsulfoximine, L-methionine sulfone, and DL-methionine sulfoxide were found to be potent inhibitors of GltS activity, yielding 10.5 values of 150, 11, and 250 ,uM, respectively. GltSs were purified from cells grown in the presence of ammonia and nitrate as sole nitrogen sources and were compared. Both yielded identical final specific activities and identical physical (UV-visible spectra, flavin, and iron-sulfur composition) and kinetic characteristics.en_US
dc.description.sponsorshipThis work was supported in part by grants PCM 7924644 and 8121902 from the National Science Foundation.en_US
dc.description.urihttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC214775/en_US
dc.format.extent9 pagesen_US
dc.genrejournal articlesen_US
dc.identifierdoi:10.13016/m27rtc-udax
dc.identifier.citationH J Schreier and R W Bernlohr, Purification and properties of glutamate synthase from Bacillus licheniformis, Journal ListJ Bacteriolv.160(2); 1984 Nov, PMC214775, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC214775/en_US
dc.identifier.urihttp://hdl.handle.net/11603/13076
dc.language.isoen_USen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Biological Sciences Department Collection
dc.relation.ispartofUMBC Department of Marine Biotechnology
dc.rightsThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
dc.subjectGlutamate synthaseen_US
dc.subjectB. Iicheniformisen_US
dc.subjectDL-methionine sulfoxideen_US
dc.subjectL-methionine sulfoneen_US
dc.subjectnucleotidesen_US
dc.titlePurification and properties of glutamate synthase from Bacillus licheniformisen_US
dc.typeTexten_US
dcterms.creatorhttps://orcid.org/0000-0003-1377-3929

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