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Literature summary for 3.1.8.2 extracted from

  • Latifi, A.M.; Karami, A.; Khodi, S.
    Efficient surface display of diisopropylfluorophosphatase (DFPase) in E. coli for biodegradation of toxic organophosphorus compounds (DFP and Cp) (2015), Appl. Biochem. Biotechnol., 177, 624-636 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 mg/l Co2+ ion increases the specific activity of the recombinant enzyme Loligo vulgaris

Application

Application Comment Organism
environmental protection the engineered bacterium, prepared with an N-terminal domain of the ice nucleation protein (InaV-N) as an anchoring motif on cell surface of expressing bacteria, can be used in the bioremediation of pesticide-contaminated environments Loligo vulgaris

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged wild-type and engineered enzymes in Escherichia coli strain BL21 (IDE3) pLysS Loligo vulgaris

Protein Variants

Protein Variants Comment Organism
additional information intracellular production of organophosphorus pesticides (OPs)-degrading enzymes or the use of native bacteria and fungi leads to a low degradation rate of OPs due to a mass transfer issue which reduces the overall catalytic efficiency. To overcome this challenge, DFPase is expressed on the surface of Escherichia coli for the first time by employing the N-terminal domain of the ice nucleation protein (InaV-N) as an anchoring motif. The recombinant DFPase is successfully located on the outer membrane and shows a significant ability for the biodegradation of diisopropylfluorophosphate (DFP) with a specific activity of 500 U/mg of wet cell weight. The recombinant cells can also degrade chlorpyrifos. No enzyme activity is measured by the fluoride ion-selective electrode in the control sample, inner membrane fraction, or cytoplasm fraction of pET-28a-InaV-N-DFPase cells. High potential of the InaV-N anchoring domain to produce an engineered bacterium that can be used in the bioremediation of pesticide-contaminated environments Loligo vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Loligo vulgaris

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates Loligo vulgaris

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
diisopropyl fluorophosphate + H2O Loligo vulgaris
-
diisopropyl phosphate + fluoride
-
?
additional information Loligo vulgaris the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs) ?
-
?

Organism

Organism UniProt Comment Textmining
Loligo vulgaris Q7SIG4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and engineered enzymes from Escherichia coli strain BL21 (IDE3) pLysS Loligo vulgaris

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
Loligo vulgaris
-
ganglion
-
Loligo vulgaris
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
additional information
-
a maximum number of 53560000 molecules of diisopropyl fluorophosphate is hydrolyzed per minute per whole cell in the presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 ppm Co2+ ion. 34.69 U/ml with substrate paraoxon, pH 7.2, 30┬░C Loligo vulgaris
382.39
-
purified recombinant engineered His-tagged InaV-N-DFPase enzyme, substrate paraoxon, pH 7.2, 30┬░C Loligo vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
diethyl-paraoxon + H2O reaction of EC 3.1.8.1, paraoxonase Loligo vulgaris diethyl phosphate + 4-nitrophenol
-
?
diisopropyl fluorophosphate + H2O
-
Loligo vulgaris diisopropyl phosphate + fluoride
-
?
additional information the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs) Loligo vulgaris ?
-
?

Subunits

Subunits Comment Organism
? x * 56847, wild-type enzyme, sequence calculation, x * 55000, recombinant wild-type enzyme, SDS-PAGE, x * 77000, recombinant engineered His-tagged InaV-N-DFPase enzyme, SDS-PAGE Loligo vulgaris

Synonyms

Synonyms Comment Organism
DFPase
-
Loligo vulgaris
diisopropylfluorophosphatase
-
Loligo vulgaris
More cf. EC 3.1.8.1 Loligo vulgaris

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
additional information
-
very low activity of whole cells at 60┬░C Loligo vulgaris
30
-
recombinant enzyme Loligo vulgaris

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Loligo vulgaris