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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

Pricing and availability is not currently available.

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.[1] The active site of the enzyme contains Fe3+.[2]

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[3] and is used to study the metabolism of protocatechuate in Rhizobiaceae[4].
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.[5] 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.[6]

Biochem/physiol Actions

Protocatechuate 3,4-Dioxygenase catalyzes the degradation of 3,4-dihydroxybenzoate (protocatechuate) into β-carboxy-cis,cis-muconate.[2]

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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G Trautwein et al.
Journal of bacteriology, 183(3), 873-881 (2001-02-24)
Protocatechuate degradation is accomplished in a multistep inducible catabolic pathway in Acinetobacter sp. strain ADP1. The induction is brought about by the transcriptional regulator PcaU in concert with the inducer protocatechuate. PcaU, a member of the new IclR family of
M Contzen et al.
Molecular microbiology, 41(1), 199-205 (2001-07-17)
The genes for a protocatechuate 3,4-dioxygenase (P34O-II) with the ability to oxidize 4-sulphocatechol were cloned from the 4-aminobenzenesulphonate(sulphanilate)-degrading bacterium Hydrogenophaga intermedia strain S1 (DSMZ 5680). Sequence comparisons of the deduced amino acid sequences of both subunits of the P34O-II from
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)
Nicole Michelotti et al.
Methods in enzymology, 475, 121-148 (2010-07-16)
Recent improvements in methods of single-particle fluorescence tracking have permitted detailed studies of molecular motion on the nanometer scale. In a quest to introduce these tools to the burgeoning field of DNA nanotechnology, we have exploited fluorescence imaging with one-nanometer
Resonance Raman studies on protocatechuate 3, 4-dioxygenase inhibitor complexes
Que J L and Epstein R M
Biochemistry, 20(9), 2545-2549 (1981)

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