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A spectrophotometric trimethylamine monooxygenase assay
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Trimethylamine monooxygenase (
Tmm
, EC-1.14.13.148) belongs to
the family of flavin-containing monooxygenases (FMOs) that oxidize
trimethylamine into trimethylamine-N-oxide (TMAO). Conventional methods
for assaying
Tmm
are accurate over a narrow range of substrate/
product concentrations. Here we report a TMAO-specific enzymatic assay
for
Tmm
using polyallylamine hydrochloride (PAHCl)-capped MnO
2
nanoparticles (PAHCl@MnO
2
). We
achieved TMAO specificity using iodoacetonitrile to remove interfering
trimethylamine. The change in the concentration of TMAO is measured by
observing the difference in the absorbance of
3,3´,5,5´-tetramethylbenzidine (TMB) at 652 nm. The assay is tolerant to
several interfering metal ions and other compounds. This method is more
reliable and easier than currently known methods. The limit of detection
(LOD) and limit of quantitation (LOQ) are 1 µM and 10 µM, respectively,
for direct TMAO measurement.
Title: A spectrophotometric trimethylamine monooxygenase assay
Description:
Trimethylamine monooxygenase (
Tmm
, EC-1.
14.
13.
148) belongs to
the family of flavin-containing monooxygenases (FMOs) that oxidize
trimethylamine into trimethylamine-N-oxide (TMAO).
Conventional methods
for assaying
Tmm
are accurate over a narrow range of substrate/
product concentrations.
Here we report a TMAO-specific enzymatic assay
for
Tmm
using polyallylamine hydrochloride (PAHCl)-capped MnO
2
nanoparticles (PAHCl@MnO
2
).
We
achieved TMAO specificity using iodoacetonitrile to remove interfering
trimethylamine.
The change in the concentration of TMAO is measured by
observing the difference in the absorbance of
3,3´,5,5´-tetramethylbenzidine (TMB) at 652 nm.
The assay is tolerant to
several interfering metal ions and other compounds.
This method is more
reliable and easier than currently known methods.
The limit of detection
(LOD) and limit of quantitation (LOQ) are 1 µM and 10 µM, respectively,
for direct TMAO measurement.
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