Abstract
A nitrogenase system that remains in the supernatant fluid after centrifuging for 3 hr at 180,000 x g can be extracted from Azotobacter vinelandii by osmotic lysis of the bacteria. This nitrogenase preparation is oxygen-labile and appears to be similar, though not identical, to that obtained from Clostridium pasteurianum. The particulate characteristic and oxygen stability of previously described preparations are likely due to the method of cell disruption, e.g., in the French pressure cell. The data support a nitrogenase model system in the intact cell in which oxygen-labile enzymes are protected from oxygen by the extensive internal membranous system which Azotobacter synthesize only when they fix nitrogen.
MeSH Terms
Acetylene/metabolism
Azotobacter/cytology,enzymology,metabolism
Cell Membrane/metabolism
Centrifugation
Microscopy, Electron
NAD/metabolism
Nitrogen Fixation
Osmosis
Oxidation-Reduction
Oxidoreductases/metabolism
Oxygen
Chemicals
NAD
Oxidoreductases
Acetylene
Oxygen
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Oppenheim J
Fisher R J
Wilson P W
Marcus L
References (13)
13 references, click to expand
-
Nitrogen fixation: cell-free system with extracts of Azotobacter.
Biochem Biophys Res Commun. 1964 Oct 14;17(3):265-71
PMID: 5862980
-
Nitrogen fixation by the bacteroid fraction of breis of soybean root nodules.
Biochim Biophys Acta. 1967 Aug 29;141(3):507-15
PMID: 6069173
-
In situ studies on N2 fixation using the acetylene reduction technique.
Proc Natl Acad Sci U S A. 1967 Nov;58(5):2071-8
PMID: 5237501
-
Is the heterocyst the site of nitrogen fixation in blue-green algae?
Nature. 1968 Nov 23;220(5169):810-2
PMID: 4880705
-
Effect of oxygen on growth of Azotobacter chroococcum in batch and continuous cultures.
J Gen Microbiol. 1968 Dec;54(3):463-73
PMID: 5709283
-
Correlation of ultrastructure in Azotobacter vinelandii with nitrogen source for growth.
J Bacteriol. 1970 Jan;101(1):286-91
PMID: 5411753
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Submicroscopic particles in extracts of Azotobacter agilis.
J Bacteriol. 1958 Mar;75(3):243-52
PMID: 13513592
-
Electron microscope study of DNA-containing plasms. II. Vegetative and mature phage DNA as compared with normal bacterial nucleoids in different physiological states.
J Biophys Biochem Cytol. 1958 Nov 25;4(6):671-8
PMID: 13610928
-
Improvements in epoxy resin embedding methods.
J Biophys Biochem Cytol. 1961 Feb;9:409-14
PMID: 13764136
-
Location of enzymes in Azotobacteragilis.
J Bacteriol. 1962 Jan;83:158-68
PMID: 14492953
-
The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.
J Cell Biol. 1963 Apr;17:208-12
PMID: 13986422
-
NITROGEN FIXATION: HYDROSULFITE AS ELECTRON DONOR WITH CELL-FREE PREPARATIONS OF AZOTOBACTER VINELANDII AND RHODOSPIRILLUM RUBRUM.
Proc Natl Acad Sci U S A. 1965 Mar;53:532-9
PMID: 14338231