Purification and some properties of nitrate reductase (EC 1.7.99.4) from Escherichia coli K12.

The Biochemical journal ·Vol. 153 ·No. 3 ·1976-03-01 ·Pages 533-41

Clegg RA

Abstract

1. Nitrate reductase was purified 134-fold from Escherichia coli K12. The purification procedure involves the release by Triton X-100 of the enzyme from the cell envelope. i. The purified enzyme exists in aqueous solution either as a monomer (mol. wt. about 220 000) or as an associated form (probably a tetramer; mol.wt. about 880 000). 3. The purified enzyme has three subunits with apparent mol.wts. of 150 000, 67000 and 65000. An additional subunit of apparent mol.wt. 20000 is present in a haem-containing fraction that is also produced by the preparative procedure described. 4. None of the enzyme subunits is present in the cell envelope of cells grown in the absence of nitrate. 5. Reversible changes in the activity of nitrate reductase in vitro with FMNH2 as reductant can be induced under circumstances which are without effect on the reduced Benzyl Viologen-NO3-activity.

MeSH Terms

Centrifugation, Density Gradient Chromatography, DEAE-Cellulose Electrophoresis, Polyacrylamide Gel Escherichia coli/enzymology Freezing Heme Microscopy, Electron Molecular Weight Nitrate Reductases/analysis,isolation & purification Nitrates/pharmacology Polyethylene Glycols

Chemicals

Nitrates Polyethylene Glycols Heme Nitrate Reductases

Authors & Affiliations

1 authors, click to expand affiliations / ORCID

Clegg R A

References (23)

23 references, click to expand

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Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj1530533

Published

1976-03-01

Pages

533-41

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1172619

Subset

External Links

PubMed source DOI: 10.1042/bj1530533 PMC Full Text View journal details

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Purification and some properties of nitrate reductase (EC 1.7.99.4) from Escherichia coli K12.

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