P8279
Protocatechuate 3,4-Dioxygenase from Pseudomonas sp.
lyophilized powder, ≥3 units/mg solid
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About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
Product Name
Protocatechuate 3,4-Dioxygenase from Pseudomonas sp., lyophilized powder, ≥3 units/mg solid
biological source
bacterial (Pseudomonas spp.)
form
lyophilized powder
specific activity
≥3 units/mg solid
mol wt
~700 kDa
shipped in
dry ice
storage temp.
−20°C
Quality Level
Analysis Note
Protein determined by biuret.
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.
General description
Protocatechuate 3,4-Dioxygenase belongs to the non-heme iron family of enzymes. The active site of the enzyme contains Fe3+.
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)
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)
Other Notes
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.
Storage Class
11 - Combustible Solids
wgk
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
G K Podila et al.
Applied and environmental microbiology, 59(8), 2717-2719 (1993-08-01)
A heterologous gene probe encoding the alpha and beta subunits of the Pseudomonas cepacia protocatechuate 3,4-dioxygenase (PCD) was used to detect its homolog in the genome of Bradyrhizobium japonicum USDA110. Three cosmid clones carrying a 2.2-kb BamHI insert showed high
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)
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
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