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Key Documents

P4591

Sigma-Aldrich

Pyruvate Oxidase from microorganisms

lyophilized powder, ≥1.5 U/mg

Synonym(s):

Pyruvate:oxygen oxidoreductase (phosphorylating)

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

form

lyophilized powder

specific activity

≥1.5 U/mg

mol wt

~260 kDa

storage temp.

−20°C

Application

Pyruvate Oxidase (PoxB) converts pyruvate directly to acetate and CO2. It is used to study pyruvate metabolism. It is used to study aerobic metabolism of bacterium, such as Lactobacillus plantarumand Streptococcus pneumoniae. Pyruvate Oxidase is used for enzymatic determination of pyruvate, GOT, and GPT in clinical analysis.
Pyruvate Oxidase from microorganisms has been used in the Amplex Red-based fluorescence assay for pyruvate. It has also been used in developing near real-time continuous detection system for pyruvate.

Biochem/physiol Actions

Pyruvate Oxidase consists of four subunits with identical molecular weights. PoxB reacts with certain aldehydes and phosphate can be replaced by arsenate. Oxygen as well as several artificial compounds can function as electron acceptors. Pyruvate Oxidase is activated by phospholipids as well as monomeric and micellar amphiphiles.
Pyruvate Oxidase oxidises pyruvate to form acetate, hydrogen peroxide and carbondioxide. Pyruvate oxidase requires flavin adenine dinucleotide (FAD), thiamine pyrophosphate (TPP) and magnesium as cofactors for its catalytic activity. Pyruvate oxidase is activated by the cofactor thiamine.

Physical properties

Isoelectric point : 4.3
Michaelis constant : 3.4 X 10-4M (Pyruvate)
Inhibitors : Fe++,Zn++,Cu++,Ag+,Hg++
Optimum pH : 5.7
Optimum temperature : 65oC
pH Stability : pH 5.7 - 6.5 (25oC, 20hr)
Thermal stability : below 45oC (pH 6.0, 15min)

Unit Definition

One unit will produce 1.0 μmole of H2O2 per min during the conversion of pyruvate and phosphate to acetylphosphate and CO2 at pH 5.7 at 37 °C.

Physical form

Lyophilized powder containing FAD and sugar as stabilizer

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Multiplexed and fully automated detection of metabolic biomarkers using microdialysis probe
Das C, et al.
Sensors and Actuators B, Chemical, 238, 633-640 (2017)
A sensitive fluorimetric assay for pyruvate
Zhu A, et al.
Analytical Biochemistry, 396(1), 146-151 (2010)
Lan-yan Zheng et al.
International journal of oral science, 3(2), 82-89 (2011-04-13)
The objective of this study was to characterize the oxygen dependent regulation of pyruvate oxidase (SpxB) gene expression and protein production in Streptococcus sanguinis (S. sanguinis). SpxB is responsible for the generation of growth-inhibiting amounts of hydrogen peroxide (H2O2) able
Amit Priyadarshi et al.
Biochemical and biophysical research communications, 380(4), 797-801 (2009-04-03)
MenD (2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexadiene-1-carboxylate) synthase belongs to the superfamily of thiamin diphosphate-dependent decarboxylases, which converts isochorismate and 2-oxoglutarate to SHCHC, pyruvate, and carbon dioxide. Here, we report the first crystal structure of apo-MenD from Escherichia coli determined in tetragonal crystal form. The
Rachel Benisty et al.
Biochimica et biophysica acta, 1801(9), 1098-1104 (2010-07-06)
FabF elongation condensing enzyme is a critical factor in determining the spectrum of products produced by the FASII pathway. Its active site contains a critical cysteine-thiol residue, which is a plausible target for oxidation by H2O2. Streptococcus pneumoniae produces exceptionally

Protocols

Assay Procedure for Pyruvate Oxidase Bacterial

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