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Volume 156, Issue 10

The obligate aerobe A3(2) synthesizes three active respiratory nitrate reductases Free

Abstract

A3(2) synthesizes three membrane-associated respiratory nitrate reductases (Nars). During aerobic growth in liquid medium the bacterium was able to reduce 50 mM nitrate stoichiometrically to nitrite. Construction and analysis of a mutant in which all three operons were deleted showed that it failed to reduce nitrate. Deletion of the gene encoding MoaA, which catalyses the first step in molybdenum cofactor biosynthesis, also prevented nitrate reduction, consistent with the Nars being molybdoenzymes. In contrast to the triple mutant, the mutant was also unable to use nitrate as sole nitrogen source, which indicates that the assimilatory nitrate reductases in are also molybdenum-dependent. Analysis of growth on solid medium demonstrated that Nar activity is present in both spores and mycelium (hypha). Development of a survival assay with the nitrate analogue chlorate revealed that wild-type spores and mycelium were sensitive to chlorate after anaerobic incubation, independent of the presence of nitrate, while both the and triple mutants were chlorate-resistant. Complementation of the triple mutant with the individual operons delivered on cosmids revealed that each operon encoded an enzyme that was synthesized and active in nitrate or chlorate reduction. The data obtained from these studies allow a tentative assignment of Nar1 activity to spores, Nar2 to spores and mycelium, and Nar3 exclusively to mycelium.

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Keyword(s): Apra, apramycin , bis-MGD, bis-molybdopterin guanine dinucleotide , Cm, chloramphenicol , Kan, kanamycin , MV, methyl viologen , Spc, spectinomycin and Strep, streptomycin
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2010-10-01
2024-04-26
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The obligate aerobe Streptomyces coelicolor A3(2) synthesizes three active respiratory nitrate reductases
Microbiology 156, 3166 (2010); https://doi.org/10.1099/mic.0.042572-0
/content/journal/micro/10.1099/mic.0.042572-0
/content/journal/micro/10.1099/mic.0.042572-0
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