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P8279

Sigma-Aldrich

Protocatechuate 3,4-Dioxygenase from Pseudomonas sp.

lyophilized powder, ≥3 units/mg solid

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About This Item

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

biological source

bacterial (Pseudomonas spp.)

Quality Level

form

lyophilized powder

specific activity

≥3 units/mg solid

mol wt

~700 kDa

shipped in

dry ice

storage temp.

−20°C

General description

Protocatechuate 3,4-Dioxygenase belongs to the non-heme iron family of enzymes. The active site of the enzyme contains Fe3+.

Application

Protocatechuate 3,4-Dioxygenase(PCD), from Pseudomonas sp., is used for the enzymatic determination of choline esterase when coupled with phydroxybenzoate hydroxylase. It is used to improve organic fluorophore-stability in single-molecule experiments and is used to study the metabolism of protocatechuate in Rhizobiaceae.
The enzyme has been used to create an oxygen scavenging system along with protocatechuate (PCA) and Trolox. The enzyme employs a nonheme iron center that catalyzes the conversion of PCA and molecular oxygen into β-carboxy-cis,cis-muconic acid, while the antioxidant Trolox suppresses slow blinking and photobleaching of cyanine dyes. It has been used in the preparation of imaging buffer along with DMB-BSA (dynein motility buffer-BSA), ATP and protocatechuate in single molecule motility assay.

Biochem/physiol Actions

Protocatechuate 3,4-Dioxygenase catalyzes the degradation of 3,4-dihydroxybenzoate (protocatechuate) into β-carboxy-cis,cis-muconate.

Physical properties

Structure : Protein with nonheme iron
Inhibitors : Ag+, Hg++, PCMB
Optimum pH : 9.0
Optimum temperature : 60−65°C
pH Stability : pH 7.0−9.0 (25°C, 72hr)
Thermal stability : below 50°C (pH 6.0, 1hr)

Unit Definition

One unit will oxidize 1.0 μmole of protocatechuate to 3-carboxy-cis,cis-muconate per min at pH 7.5 at 37 °C.

Physical form

Supplied as lyophilized powder.

Analysis Note

Protein determined by biuret.

inhibitor

Product No.
Description
Pricing

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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Mindy I Davis et al.
Journal of the American Chemical Society, 124(4), 602-614 (2002-01-24)
The geometric and electronic structure of the high-spin ferric active site of protocatechuate 3,4-dioxygenase (3,4-PCD) has been examined by absorption (Abs), circular dichroism (CD), magnetic CD (MCD), and variable-temperature-variable-field (VTVH) MCD spectroscopies. Density functional (DFT) and INDO/S-CI molecular orbital calculations
A Buchan et al.
Applied and environmental microbiology, 67(12), 5801-5809 (2001-11-28)
Degradation of lignin-related aromatic compounds is an important ecological process in the highly productive salt marshes of the southeastern United States, yet little is known about the mediating organisms or their catabolic pathways. Here we report the diversity of a
Colin Echeverría Aitken et al.
Biophysical journal, 94(5), 1826-1835 (2007-10-09)
The application of single-molecule fluorescence techniques to complex biological systems places demands on the performance of single fluorophores. We present an enzymatic oxygen scavenging system for improved dye stability in single-molecule experiments. We compared the previously described protocatechuic acid/protocatechuate-3,4-dioxygenase system
Resonance Raman studies on protocatechuate 3, 4-dioxygenase inhibitor complexes
Que J L and Epstein R M
Biochemistry, 20(9), 2545-2549 (1981)
Brevibacterium fuscum protocatechuate 3, 4-dioxygenase. Purification, crystallization, and characterization.
Whittaker J W, et al.
The Journal of Biological Chemistry, 259(7), 4466-4475 (1984)

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