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PMID: 11844760 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Periplasmic nitrate reductase (NapABC enzyme) supports anaerobic respiration by Escherichia coli K-12.

Journal of bacteriology ·Vol. 184 ·No. 5 ·2002-03-00 ·Pages 1314-23

Stewart V, Lu Y, Darwin AJ

Abstract

Periplasmic nitrate reductase (NapABC enzyme) has been characterized from a variety of proteobacteria, especially Paracoccus pantotrophus. Whole-genome sequencing of Escherichia coli revealed the structural genes napFDAGHBC, which encode NapABC enzyme and associated electron transfer components. E. coli also expresses two membrane-bound proton-translocating nitrate reductases, encoded by the narGHJI and narZYWV operons. We measured reduced viologen-dependent nitrate reductase activity in a series of strains with combinations of nar and nap null alleles. The napF operon-encoded nitrate reductase activity was not sensitive to azide, as shown previously for the P. pantotrophus NapA enzyme. A strain carrying null alleles of narG and narZ grew exponentially on glycerol with nitrate as the respiratory oxidant (anaerobic respiration), whereas a strain also carrying a null allele of napA did not. By contrast, the presence of napA+ had no influence on the more rapid growth of narG+ strains. These results indicate that periplasmic nitrate reductase, like fumarate reductase, can function in anaerobic respiration but does not constitute a site for generating proton motive force. The time course of phi(napF-lacZ) expression during growth in batch culture displayed a complex pattern in response to the dynamic nitrate/nitrite ratio. Our results are consistent with the observation that phi(napF-lacZ) is expressed preferentially at relatively low nitrate concentrations in continuous cultures (H. Wang, C.-P. Tseng, and R. P. Gunsalus, J. Bacteriol. 181:5303-5308, 1999). This finding and other considerations support the hypothesis that NapABC enzyme may function in E. coli when low nitrate concentrations limit the bioenergetic efficiency of nitrate respiration via NarGHI enzyme.

MeSH Terms
Anaerobiosis Bacterial Proteins/genetics,metabolism Carbon/metabolism Culture Media Escherichia coli/enzymology,genetics,growth & development Escherichia coli Proteins Gene Expression Regulation, Bacterial Kinetics Nitrate Reductase Nitrate Reductases/genetics,metabolism Nitrates/metabolism Nitrites/metabolism Oxygen Consumption Periplasm/enzymology
Chemicals
Bacterial Proteins Culture Media Escherichia coli Proteins Nitrates Nitrites Carbon Nitrate Reductases Nitrate Reductase napA protein, E coli narZ protein, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Stewart Valley
Section of Microbiology, University of California, O)ne Shields Avenue, Davis, CA 95616-8665, USA. [email protected]
Lu Yiran
Darwin Andrew J
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.5.1314-1323.2002
Email
[email protected]
Published
2002-03-00
Pages
1314-23
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC134854
Subset
IM
Grants
NIGMS NIH HHS · R01 GM036877 · United States
NIGMS NIH HHS · GM 36877 · United States
External Links
PubMed source DOI: 10.1128/JB.184.5.1314-1323.2002 PMC Full Text View journal details
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