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Pyruvate Oxidation

Pyruvate Oxidation

After glycolysis, the charged pyruvate molecules enter the mitochondria via active transport and undergo three enzymatic reactions. These reactions ensure that pyruvate can enter the next metabolic pathway so that energy stored in the pyruvate molecules can be harnessed by the cells.
First, the enzyme pyruvate dehydrogenase removes the carboxyl group from pyruvate and releases it as carbon dioxide. The stripped molecule is then oxidized and releases electrons, which are then picked up by NAD+...

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Updated: Jun 20, 2026

Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

Published on: October 15, 2018

A sensitive fluorimetric assay for pyruvate.

Aiping Zhu¹, Roberto Romero, Howard R Petty

¹Department of Ophthalmology and Visual Sciences, University of Michigan Medical School, Ann Arbor, MI 48105, USA.

Analytical Biochemistry

|September 15, 2009

Summary

A new fluorimetric assay accurately measures pyruvate concentrations using pyruvate oxidase and Amplex Red. This sensitive method detects pyruvate at 5 nM, useful for intracellular and extracellular measurements.

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Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

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Area of Science:

Biochemistry
Analytical Chemistry
Biotechnology

Background:

Pyruvate is a key metabolite in cellular respiration and metabolic pathways.
Accurate measurement of pyruvate is crucial for understanding metabolic states.
Existing pyruvate assays may lack sensitivity or specificity.

Purpose of the Study:

To develop a sensitive and specific fluorimetric assay for pyruvate determination.
To optimize assay conditions for accurate pyruvate quantification.
To demonstrate the assay's utility in biological samples.

Main Methods:

Utilized pyruvate oxidase for pyruvate oxidation.
Employed Amplex Red to detect hydrogen peroxide produced during oxidation.
Optimized reaction buffer pH, enzyme and dye concentrations, and incubation time.
Validated the assay for measuring intracellular and extracellular pyruvate.

Main Results:

Developed a fluorimetric assay based on pyruvate oxidase and Amplex Red.
Resorufin fluorescence is stoichiometrically generated from pyruvate.
Optimized assay parameters ensure reliable performance.
Achieved a limit of detection of 5 nM for pyruvate.
Demonstrated accurate pyruvate measurement in biological samples.

Conclusions:

The developed fluorimetric assay is sensitive and specific for pyruvate.
This assay provides a reliable method for quantifying pyruvate in various biological contexts.
The low limit of detection allows for precise measurements of pyruvate levels.