Effect of carbon substrate and aeration on nitrate reduction and expression of the periplasmic and membrane-bound nitrate reductases in carbon-limited continuous cultures of Paracoccus denitrificans Pd1222

Heather J. Sears, Stephen Spiro, David J. Richardson

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39 Citations (Scopus)

Abstract

The expression and in situ activity of the membrane-bound and periplasmic nitrate reductases have been assayed in Paracoccus denitrificans Pd1222 grown under a range of aeration regimes in malate-limited or butyrate-limited chemostat cultures. In butyrate-limited cultures the expression of periplasmic nitrate reductase and the rate of in situ nitrate reduction were high at all oxygen concentrations measured between 0% and 100% air saturation. By contrast, in malate-limited cultures expression of the periplasmic nitrate reductase was low at 80-100% air saturation but increased to a maximum between 20% and 50% air saturation. Aerobic nitrate reduction was much higher in butyrate-limited than in malate-limited cultures, demonstrating a significant role for this process during butyrate metabolism. The rate of nitrate respiration increased in both the malate- and butyrate-limited cultures as aerobic metabolism switched completely to anaerobic metabolism. Expression of the membrane-bound nitrate reductase could be detected in butyrate-limited chemostat cultures maintained at an oxygen level of 100% air saturation. No membrane-bound nitrate reductase was detectable under similar conditions in malate-limited cultures but expression was detected at oxygen concentrations of 50% air saturation and below. Taken together, the results show that the nature of the carbon substrate and oxygen concentration can both influence expression of the periplasmic and membrane-bound nitrate reductases. The conditions under which expression of the periplasmic nitrate reductase and aerobic nitrate respiration are maximal can be rationalized in terms of a role for the periplasmic nitrate reductase in dissipating excess reductant generated during oxidative metabolism of reduced carbon substrates.

Original languageEnglish
Pages (from-to)3767-3774
Number of pages8
JournalMicrobiology
Volume143
Issue number12
DOIs
Publication statusPublished - 1 Dec 1997
Externally publishedYes

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Nitrate Reductases
Paracoccus denitrificans
Nitrate Reductase
Butyrates
Nitrates
Carbon
Air
Membranes
Oxygen
Anaerobiosis
Reducing Agents
Respiratory Rate
Respiration
malic acid

Keywords

  • Aerobic nitrate respiration
  • Denitrification
  • Nitrate reductase
  • Paracoccus denitrificans
  • Redox balancing

ASJC Scopus subject areas

  • Microbiology

Cite this

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title = "Effect of carbon substrate and aeration on nitrate reduction and expression of the periplasmic and membrane-bound nitrate reductases in carbon-limited continuous cultures of Paracoccus denitrificans Pd1222",
abstract = "The expression and in situ activity of the membrane-bound and periplasmic nitrate reductases have been assayed in Paracoccus denitrificans Pd1222 grown under a range of aeration regimes in malate-limited or butyrate-limited chemostat cultures. In butyrate-limited cultures the expression of periplasmic nitrate reductase and the rate of in situ nitrate reduction were high at all oxygen concentrations measured between 0{\%} and 100{\%} air saturation. By contrast, in malate-limited cultures expression of the periplasmic nitrate reductase was low at 80-100{\%} air saturation but increased to a maximum between 20{\%} and 50{\%} air saturation. Aerobic nitrate reduction was much higher in butyrate-limited than in malate-limited cultures, demonstrating a significant role for this process during butyrate metabolism. The rate of nitrate respiration increased in both the malate- and butyrate-limited cultures as aerobic metabolism switched completely to anaerobic metabolism. Expression of the membrane-bound nitrate reductase could be detected in butyrate-limited chemostat cultures maintained at an oxygen level of 100{\%} air saturation. No membrane-bound nitrate reductase was detectable under similar conditions in malate-limited cultures but expression was detected at oxygen concentrations of 50{\%} air saturation and below. Taken together, the results show that the nature of the carbon substrate and oxygen concentration can both influence expression of the periplasmic and membrane-bound nitrate reductases. The conditions under which expression of the periplasmic nitrate reductase and aerobic nitrate respiration are maximal can be rationalized in terms of a role for the periplasmic nitrate reductase in dissipating excess reductant generated during oxidative metabolism of reduced carbon substrates.",
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T1 - Effect of carbon substrate and aeration on nitrate reduction and expression of the periplasmic and membrane-bound nitrate reductases in carbon-limited continuous cultures of Paracoccus denitrificans Pd1222

AU - Sears, Heather J.

AU - Spiro, Stephen

AU - Richardson, David J.

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N2 - The expression and in situ activity of the membrane-bound and periplasmic nitrate reductases have been assayed in Paracoccus denitrificans Pd1222 grown under a range of aeration regimes in malate-limited or butyrate-limited chemostat cultures. In butyrate-limited cultures the expression of periplasmic nitrate reductase and the rate of in situ nitrate reduction were high at all oxygen concentrations measured between 0% and 100% air saturation. By contrast, in malate-limited cultures expression of the periplasmic nitrate reductase was low at 80-100% air saturation but increased to a maximum between 20% and 50% air saturation. Aerobic nitrate reduction was much higher in butyrate-limited than in malate-limited cultures, demonstrating a significant role for this process during butyrate metabolism. The rate of nitrate respiration increased in both the malate- and butyrate-limited cultures as aerobic metabolism switched completely to anaerobic metabolism. Expression of the membrane-bound nitrate reductase could be detected in butyrate-limited chemostat cultures maintained at an oxygen level of 100% air saturation. No membrane-bound nitrate reductase was detectable under similar conditions in malate-limited cultures but expression was detected at oxygen concentrations of 50% air saturation and below. Taken together, the results show that the nature of the carbon substrate and oxygen concentration can both influence expression of the periplasmic and membrane-bound nitrate reductases. The conditions under which expression of the periplasmic nitrate reductase and aerobic nitrate respiration are maximal can be rationalized in terms of a role for the periplasmic nitrate reductase in dissipating excess reductant generated during oxidative metabolism of reduced carbon substrates.

AB - The expression and in situ activity of the membrane-bound and periplasmic nitrate reductases have been assayed in Paracoccus denitrificans Pd1222 grown under a range of aeration regimes in malate-limited or butyrate-limited chemostat cultures. In butyrate-limited cultures the expression of periplasmic nitrate reductase and the rate of in situ nitrate reduction were high at all oxygen concentrations measured between 0% and 100% air saturation. By contrast, in malate-limited cultures expression of the periplasmic nitrate reductase was low at 80-100% air saturation but increased to a maximum between 20% and 50% air saturation. Aerobic nitrate reduction was much higher in butyrate-limited than in malate-limited cultures, demonstrating a significant role for this process during butyrate metabolism. The rate of nitrate respiration increased in both the malate- and butyrate-limited cultures as aerobic metabolism switched completely to anaerobic metabolism. Expression of the membrane-bound nitrate reductase could be detected in butyrate-limited chemostat cultures maintained at an oxygen level of 100% air saturation. No membrane-bound nitrate reductase was detectable under similar conditions in malate-limited cultures but expression was detected at oxygen concentrations of 50% air saturation and below. Taken together, the results show that the nature of the carbon substrate and oxygen concentration can both influence expression of the periplasmic and membrane-bound nitrate reductases. The conditions under which expression of the periplasmic nitrate reductase and aerobic nitrate respiration are maximal can be rationalized in terms of a role for the periplasmic nitrate reductase in dissipating excess reductant generated during oxidative metabolism of reduced carbon substrates.

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KW - Denitrification

KW - Nitrate reductase

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