Characterization of the expression and activity of the periplasmic nitrate reductase of Paracoccus pantotrophus in chemostat cultures

M J K Ellington, G Sawers, H J Sears, S Spiro, D J Richardson, S J Ferguson

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The periplasmic nitrate reductase (Nap) from Paracoccus pantotrophus has a role in cellular redox balancing. Previously, transcription from the nap promoter in P. pantotrophus was shown to be responsive to the oxidation state of the carbon substrate. During batch culture, expression was higher during growth on reduced substrates such as butyrate compared to more oxidized substrates such as succinate. In the present study the effect of growth rate on nap expression in succinate-, acetate- and butyrate-limited chemostat cultures was investigated. In all three cases transcription from the nap promoter and Nap enzyme activity showed a strong correlation. At the fastest growth rates tested for the three substrates nap expression and Nap activity were highest when growth occurred on the most reduced substrate (butyrate > acetate > succinate). However, in all three cases a bell-shaped pattern of expression was observed as a function of growth rate, with the highest levels of nap expression and Nap activity being observed at intermediate growth rates. This effect was most pronounced on succinate, where an approximately fivefold variation was observed, and at intermediate dilution rates nap expression and Nap activity were comparable on all three carbon substrates. Analysis of mRNA prepared from the succinate-grown cultures revealed that different transcription initiation start sites for the nap operon were utilized as the growth rate changed. This study establishes a new regulatory feature of nap expression in P. pantotrophus that occurs at the level of transcription in response to growth rate in carbon-limited cultures.

Original languageEnglish
Pages (from-to)1533-1540
Number of pages8
Journal BMC Microbiology
Volume149
Issue number6
DOIs
Publication statusPublished - 1 Jun 2003

Fingerprint

Paracoccus pantotrophus
Nitrate Reductase
Succinic Acid
Growth
Butyrates
Carbon
Transcription Initiation Site
Acetates
Batch Cell Culture Techniques
Operon
Oxidation-Reduction

Keywords

  • Acetic Acid
  • Aerobiosis
  • Butyrates
  • Culture Media
  • Gene Expression
  • Genes, Bacterial
  • Nitrate Reductase
  • Nitrate Reductases
  • Nitrates
  • Nitrites
  • Paracoccus
  • Periplasm
  • Promoter Regions, Genetic
  • Succinic Acid
  • Transcription, Genetic
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Characterization of the expression and activity of the periplasmic nitrate reductase of Paracoccus pantotrophus in chemostat cultures. / Ellington, M J K; Sawers, G; Sears, H J; Spiro, S; Richardson, D J; Ferguson, S J.

In: BMC Microbiology , Vol. 149, No. 6, 01.06.2003, p. 1533-1540.

Research output: Contribution to journalArticle

Ellington, M J K ; Sawers, G ; Sears, H J ; Spiro, S ; Richardson, D J ; Ferguson, S J. / Characterization of the expression and activity of the periplasmic nitrate reductase of Paracoccus pantotrophus in chemostat cultures. In: BMC Microbiology . 2003 ; Vol. 149, No. 6. pp. 1533-1540.
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T1 - Characterization of the expression and activity of the periplasmic nitrate reductase of Paracoccus pantotrophus in chemostat cultures

AU - Ellington, M J K

AU - Sawers, G

AU - Sears, H J

AU - Spiro, S

AU - Richardson, D J

AU - Ferguson, S J

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N2 - The periplasmic nitrate reductase (Nap) from Paracoccus pantotrophus has a role in cellular redox balancing. Previously, transcription from the nap promoter in P. pantotrophus was shown to be responsive to the oxidation state of the carbon substrate. During batch culture, expression was higher during growth on reduced substrates such as butyrate compared to more oxidized substrates such as succinate. In the present study the effect of growth rate on nap expression in succinate-, acetate- and butyrate-limited chemostat cultures was investigated. In all three cases transcription from the nap promoter and Nap enzyme activity showed a strong correlation. At the fastest growth rates tested for the three substrates nap expression and Nap activity were highest when growth occurred on the most reduced substrate (butyrate > acetate > succinate). However, in all three cases a bell-shaped pattern of expression was observed as a function of growth rate, with the highest levels of nap expression and Nap activity being observed at intermediate growth rates. This effect was most pronounced on succinate, where an approximately fivefold variation was observed, and at intermediate dilution rates nap expression and Nap activity were comparable on all three carbon substrates. Analysis of mRNA prepared from the succinate-grown cultures revealed that different transcription initiation start sites for the nap operon were utilized as the growth rate changed. This study establishes a new regulatory feature of nap expression in P. pantotrophus that occurs at the level of transcription in response to growth rate in carbon-limited cultures.

AB - The periplasmic nitrate reductase (Nap) from Paracoccus pantotrophus has a role in cellular redox balancing. Previously, transcription from the nap promoter in P. pantotrophus was shown to be responsive to the oxidation state of the carbon substrate. During batch culture, expression was higher during growth on reduced substrates such as butyrate compared to more oxidized substrates such as succinate. In the present study the effect of growth rate on nap expression in succinate-, acetate- and butyrate-limited chemostat cultures was investigated. In all three cases transcription from the nap promoter and Nap enzyme activity showed a strong correlation. At the fastest growth rates tested for the three substrates nap expression and Nap activity were highest when growth occurred on the most reduced substrate (butyrate > acetate > succinate). However, in all three cases a bell-shaped pattern of expression was observed as a function of growth rate, with the highest levels of nap expression and Nap activity being observed at intermediate growth rates. This effect was most pronounced on succinate, where an approximately fivefold variation was observed, and at intermediate dilution rates nap expression and Nap activity were comparable on all three carbon substrates. Analysis of mRNA prepared from the succinate-grown cultures revealed that different transcription initiation start sites for the nap operon were utilized as the growth rate changed. This study establishes a new regulatory feature of nap expression in P. pantotrophus that occurs at the level of transcription in response to growth rate in carbon-limited cultures.

KW - Acetic Acid

KW - Aerobiosis

KW - Butyrates

KW - Culture Media

KW - Gene Expression

KW - Genes, Bacterial

KW - Nitrate Reductase

KW - Nitrate Reductases

KW - Nitrates

KW - Nitrites

KW - Paracoccus

KW - Periplasm

KW - Promoter Regions, Genetic

KW - Succinic Acid

KW - Transcription, Genetic

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1099/mic.0.26277-0

DO - 10.1099/mic.0.26277-0

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VL - 149

SP - 1533

EP - 1540

JO - BMC Microbiology

JF - BMC Microbiology

SN - 1471-2180

IS - 6

ER -