In vivo enzymology: A deuterium NMR study of formaldehyde dismutase in Pseudomonas putida F61a and Staphylococcus aureus

Ralph P. Mason, Jeremy K M Sanders

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

High-resolution deuterium NMR spectroscopy has been used to follow the detoxifying metabolism of [D2] formaldehyde in vivo in several bacterial species. Production of [D2]methanol in Escherichia coli confirms that the oxidation and reduction pathways of metabolism are independent in this organism. Efficient production of equimolar quantities of [D]formate and [D3]methanol in Pseudomonas putida F61a and Staphylococcus aureus implicates a formaldehyde dismutase, or "cannizzarase", activity. These observations imply that the unusual formaldehyde resistance in P. putida F61a is a direct result of efficient dismutation acting as a route for detoxification. Cross-dismutation experiments yield an enzymic kinetic isotope effect of ca. 4 for H vs D transfer and a similar spectrum of substrate specificity to the isolated enzyme. [D]benzyl alcohol produced by cross-dismutation of [D2] formaldehyde and benzaldehyde in P. putida is demonstrated to have the R configuration by a novel deuterium NMR assay. Additionally, S. aureus produces methyl formate as a product of formaldehyde detoxification, apparently by oxidizing the methanol hemiacetal of formaldehyde.

Original languageEnglish (US)
Pages (from-to)2160-2168
Number of pages9
JournalBiochemistry
Volume28
Issue number5
StatePublished - 1989

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Pseudomonas putida
Deuterium
Formaldehyde
Staphylococcus aureus
Nuclear magnetic resonance
Methanol
Detoxification
formic acid
Metabolism
Benzyl Alcohol
Substrate Specificity
Isotopes
Escherichia coli
Nuclear magnetic resonance spectroscopy
Oxidation-Reduction
Assays
Magnetic Resonance Spectroscopy
formaldehyde dismutase
Oxidation
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

In vivo enzymology : A deuterium NMR study of formaldehyde dismutase in Pseudomonas putida F61a and Staphylococcus aureus. / Mason, Ralph P.; Sanders, Jeremy K M.

In: Biochemistry, Vol. 28, No. 5, 1989, p. 2160-2168.

Research output: Contribution to journalArticle

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