scholarly journals A respiratory nitrate reductase active exclusively in resting spores of the obligate aerobeStreptomyces coelicolor A3(2)

2013 ◽  
Vol 89 (6) ◽  
pp. 1259-1273 ◽  
Author(s):  
Marco Fischer ◽  
Dörte Falke ◽  
R. Gary Sawers
Keyword(s):  
Nitrate Reductase ◽  
Resting Spores ◽  
Respiratory Nitrate Reductase

scholarly journals Activity of Spore-Specific Respiratory Nitrate Reductase 1 ofStreptomyces coelicolorA3(2) Requires a Functional Cytochromebcc-aa3Oxidase Supercomplex

2019 ◽  
Vol 201 (11) ◽  
Author(s):  
Dörte Falke ◽  
Bianca Biefel ◽  
Alexander Haase ◽  
Stefan Franke ◽  
Marco Fischer ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Nitrate Reductase ◽  
Nitrate Reduction ◽  
Streptomyces Coelicolor ◽  
Nitrate Respiration ◽  
Nitrate Transport ◽  
Developmental Cycle ◽  
Content Type ◽  
Resting Spores ◽  
Respiratory Nitrate Reductase

ABSTRACTSpores have strongly reduced metabolic activity and are produced during the complex developmental cycle of the actinobacteriumStreptomyces coelicolor. Resting spores can remain viable for decades, yet little is known about how they conserve energy. It is known, however, that they can reduce either oxygen or nitrate using endogenous electron sources.S. coelicoloruses either a cytochromebdoxidase or a cytochromebcc-aa3oxidase supercomplex to reduce oxygen, while nitrate is reduced by Nar-type nitrate reductases, which typically oxidize quinol directly. Here, we show that in resting spores the Nar1 nitrate reductase requires a functionalbcc-aa3supercomplex to reduce nitrate. Mutants lacking the completeqcr-ctagenetic locus encoding thebcc-aa3supercomplex showed no Nar1-dependent nitrate reduction. Recovery of Nar1 activity was achieved by genetic complementation but only when the completeqcr-ctalocus was reintroduced to the mutant strain. We could exclude that the dependence on the supercomplex for nitrate reduction was via regulation of nitrate transport. Moreover, the catalytic subunit, NarG1, of Nar1 was synthesized in theqcr-ctamutant, ruling out transcriptional control. Constitutive synthesis of Nar1 in mycelium revealed that the enzyme was poorly active in this compartment, suggesting that the Nar1 enzyme cannot act as a typical quinol oxidase. Notably, nitrate reduction by the Nar2 enzyme, which is active in growing mycelium, was not wholly dependent on thebcc-aa3supercomplex for activity. Together, our data suggest that Nar1 functions together with the proton-translocatingbcc-aa3supercomplex to increase the efficiency of energy conservation in resting spores.IMPORTANCEStreptomyces coelicolorforms spores that respire with either oxygen or nitrate, using only endogenous electron donors. This helps maintain a membrane potential and, thus, viability. Respiratory nitrate reductase (Nar) usually receives electrons directly from reduced quinone species; however, we show that nitrate respiration in spores requires a respiratory supercomplex comprising cytochromebccoxidoreductase andaa3oxidase. Our findings suggest that the Nar1 enzyme in theS. coelicolorspore functions together with the proton-translocatingbcc-aa3supercomplex to help maintain the membrane potential more efficiently. Dissecting the mechanisms underlying this survival strategy is important for our general understanding of bacterial persistence during infection processes and of how bacteria might deal with nutrient limitation in the natural environment.

RNA-seq analyses reveal insights into the function of respiratory nitrate reductase of the diazotroph Herbaspirillum seropedicae

2016 ◽  
Vol 18 (8) ◽  
pp. 2677-2688 ◽  
Author(s):  
Paloma Bonato ◽  
Marcelo B. Batista ◽  
Doumit Camilios-Neto ◽  
Vânia C. S. Pankievicz ◽  
Michelle Z. Tadra-Sfeir ◽  
...  
Keyword(s):  
Nitrate Reductase ◽  
Rna Seq ◽  
Herbaspirillum Seropedicae ◽  
Respiratory Nitrate Reductase

Mass spectrometry identification of membrane-bound respiratory nitrate reductase from Bradyrhizobium sp. (Lupinus).

2008 ◽  
Vol 55 (4) ◽  
pp. 753-760 ◽  
Author(s):  
Władysław Polcyn
Keyword(s):  
Mass Spectrometry ◽  
Nitrate Reductase ◽  
Biochemical Properties ◽  
Validation Criteria ◽  
Sds Page ◽  
Mass Spectrometry Identification ◽  
Bradyrhizobium Sp ◽  
Rhizobial Species ◽  
Membrane Bound ◽  
Respiratory Nitrate Reductase

Respiratory nitrate reductase (NR) from Bradyrhizobium sp. (Lupinus) USDA 3045 has biochemical properties of the membrane-bound NR type. However, in the completely sequenced rhizobium genomes only genes for the periplasmic type of dissimilatory NR were found. Therefore purification and identification of the enzyme by tandem mass spectrometry (MS/MS) was undertaken. MS/MS spectra representing 149 unique tryptic peptides derived from purified 137-kDa subunit matched the NCBInr-deposited NarG sequences. MS/MS sequencing of two other SDS/PAGE bands (65- and 59-kDa) identified them as derivatives of the NarH-type protein. Applying additional validation criteria, 73% of the sequence of the NarG subunit (902 aa) and 52% of NarH sequence (266 aa) was assembled (UniProt KB acc. no. P85097 and P85098). This is the first unambiguous identification of an active NarGH-like NR in rhizobia. Moreover, arguments are provided here for the existence of a functional enzyme of this type also among other rhizobial species, basing on immunoblot screening and the presence of membrane-associated NR-active electrophoretic forms.

scholarly journals Investigation by electron paramagnetic resonance spectroscopy of the molybdenum centre of respiratory nitrate reductase from Paracoccus denitrificans

1988 ◽  
Vol 252 (3) ◽  
pp. 925-926 ◽  
Author(s):  
N Turner ◽  
A L Ballard ◽  
R C Bray ◽  
S Ferguson
Keyword(s):  
Escherichia Coli ◽  
Nitrate Reductase ◽  
Paracoccus Denitrificans ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Resonance Spectroscopy ◽  
Species Comparison ◽  
Paramagnetic Resonance ◽  
Electron Paramagnetic ◽  
Respiratory Nitrate Reductase ◽  
Active Centres

The molybdenum centre of respiratory nitrate reductase from Paracoccus denitrificans has been investigated by e.p.r. spectroscopy of Mo(V). In common with the centres of the analogous enzymes from Escherichia coli and Pseudomonas aeruginosa, it undergoes a pH- and anion-dependent transition between two different e.p.r. signal-giving species. Comparison of the relevant e.p.r. parameters extracted with the help of computer simulations reveals ligation of the metal in the active centres of the three enzymes to be identical.

Sign in / Sign up

Export Citation Format

Share Document