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Hydroxylamine metabolism in Pseudomonas PB16: involvement of a novel hydroxylamine oxidoreductase

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Abstract

Pseudomonas strain PB16, a Gram-negative heterotrophic nitrifying bacterium closely related to Pseudomonas azalaica on the basis of 16 S rDNA analysis, was able to use hydroxylamine as an additional energy source during growth in acetate limited chemostat cultures giving an increased biomass yield. In aerobically growing cells of Pseudomonas PB16 only 50% of supplemented hydroxylamine could be recovered as nitrite. In addition to nitrite, N2O could be detected in the chemostat off-gas, indicating combined heterotrophic nitrification and aerobic denitrification. The maximum specific hydroxylamine oxidizing activity observed was 450 nmol per min per mg dry weight, with a Ks of approximately 40 µm. Upon addition of hydroxylamine to the medium, Pseudomonas PB16 induced a soluble 132 KDa dimeric hydroxylamine oxidoreductase. The enzyme had a pH optimum of 9, and did not contain spectroscopic features typical for cytochromes, which is in contrast to hydroxylamine oxidoreductases fou nd in autotrophic bacteria.

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References

  • Arciero DM & Hooper AB (1993) Hydroxylamine oxidoreductase from Nitrosomonas europaea is a multimer of an octo-heme subunit. J.Biol. Chem. 268: 14645–14654

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254

    Google Scholar 

  • Castignetti D, Palutsis D & Turley J (1990) An examination of proton translocation and energy conservation during heterotrophic nitrification. FEMS Microbiol. Lett. 66: 175–182.

    Google Scholar 

  • de Bruijn P, van de Graaf AA, Jetten MSM, Robertson LA & Kuenen JG (1995) Growth of Nitrosomonas europeae on hydroxye-amine. FEMS Microbiol. Lett 125: 179–184.

    Google Scholar 

  • Ferguson S (1994) Denitrification and its control. A. van leeuwenhoek 66:89–110

    Google Scholar 

  • Frear, DS & Burrell RC (1955) Spectrophotometric method for determining hydroxylamine reductase activity in higher plants. Anal. Chem. 27: 1664–1665

    Google Scholar 

  • Griess-Romijn-van Eck (1966) Physiological and chemical tests for drinking water, NEN 1056, IV-2. Nederlands Normalisatie-Instituut Rijswijk

  • Hooper AB, Maxwell PC & Terry KR (1978) Hydroxylamine oxidoreductase from Nitrosomonas. Biochemistry17: 2984–2989

    Google Scholar 

  • ICSB (1982) Opinion 54: Rejection of the species Pseudomonas denitrificans. Int. J. Syst. Bacteriol. 32: 466.

    Google Scholar 

  • Kuenen JG & Robertson LA (1994) Combined nitrification and denitrification processes. FEMS Microbiol. Rev. 15: 109–117.

    Google Scholar 

  • Kurokawa M, Fukumori Y & Yamanaka T (1985) A hydroxylamine-Cytochrome C reductase occurs in the heterotrophic nitrifier Arthrobacter globiformis. Plant Cell Physiol. 26: 1439–1442.

    Google Scholar 

  • Robertson LA & Kuenen JG (1988) Heterotrophic nitrification in Thiosphaera pantotropa Oxygen uptake and enzyme studies. J. Gen. Microbiol. 134: 857–863.

    Google Scholar 

  • Robertson LA, Cornelisse R, de Vos P, Hadioetomo R & Kuenen JG (1989) Aerobic denitrification in various heterotrophic nitrifiers. A. van Leeuwenhoek 56, 289–299.

    Google Scholar 

  • Robertson LA & Kuenen JG (1990) Combined heterotrophic nitrification and aerobic denitrification in Thiosphaera pantotropha and other bacteria. A. van Leeuwenhoek 57, 139–152

    Google Scholar 

  • Stouthamer AH (1995) Introduction to the metabolism of inorganic nitrogen compounds. Beijerinck Centennial (Scheffers WA & van Dijken JP Eds) pp 120–121, Delft University Press, Delft, the Netherlands

    Google Scholar 

  • Tokuyama T, Takahaski R & Tomita Y (1988) Oxidation of hydroxylamine by Pseudomonas putida. Hakkokogaku 66, 401–404

    Google Scholar 

  • van Niel EWJ, Braber KJ, Robertson LA & Kuenen JG (1992) Heterotrophic nitrification and aerobic denitrification in Alcaligenes faecalis strain TUD. A van Leeuwenhoek 62: 231–237

    Google Scholar 

  • Wehrfritz JM, Reilly A, Spiro S & Richarson DJ (1993) Purification of hydroxylamine oxidase from Thiosphaera pantotropha. FEBS Lett. 335, 246–250

    Google Scholar 

  • Zahn JA, Duncan C & Dispirito AA (1994) Oxidation of hydroxylamine by cytochrome P-460 of the obligate methylotroph Methylococcus capsulatus Bath. J. Bacteriol. 176, 5879–5887

    Google Scholar 

Download references

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Authors and Affiliations

  1. Kluyver Laboratory for Biotechnology, Delft University of Technology, Julianalaan 67, NL, 2628 BC, Delft, The Netherlands

    Mike S.M. Jetten, Mike S.M. Jetten, Peter de Bruijn & J. Gijs Kuenen

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  1. Mike S.M. Jetten
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  2. Peter de Bruijn
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  3. J. Gijs Kuenen
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Jetten, M.S., de Bruijn, P. & Kuenen, J.G. Hydroxylamine metabolism in Pseudomonas PB16: involvement of a novel hydroxylamine oxidoreductase. Antonie Van Leeuwenhoek 71, 69–74 (1997). https://doi.org/10.1023/A:1000145617904

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  • DOI: https://doi.org/10.1023/A:1000145617904

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