Phosphotriesterase from Pseudomonas diminuta

Enzyme Description

Species

Extremophile

EC Number

3.1.8.1 ↗

Sequence

Length: 365 amino acids

MQTRRVVLKSAAAAGTLLGGLAGCASVAGSIGTGDRINTVRGPITISEAGFTLTHEHICGSSAGFLRAWPEFFGSRKALAEKAVRGLRRARAAGVRTIVDVSTFDIGRDVSLLAEVSRAADVHIVAATGLWFDPPLSMRLRSVEELTQFFLREIQYGIEDTGIRAGIIKVATTGKATPFQELVLKAAARASLATGVPVTTHTAASQRDGEQQAAIFESEGLSPSRVCIGHSDDTDDLSYLTALAARGYLIGLDHIPHSAIGLEDNASASALLGIRSWQTRALLIKALIDQGYMKQILVSNDWLFGFSSYVTNIMDVMDRVNPDGMAFIPLRVIPFLREKGVPQETLAGITVTNPARFLSPTLRAS

Keith E. LeJeune et al. (1997) — Dramatically stabilized phosphotriesterase—polymers for nerve agent degradation

Biotechnology and Bioengineering · doi:10.1002/(SICI)1097-0290(19970420)54:2%3C105::AID-BIT2%3E3.0.CO;2-P ↗ · Activity - Classical Stability - Incubation Stability - Half-life

22 measurements ▼

Database ID

UniProt: P0A434 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Purified, bacterium Pseudomonas diminuta

Experimental Data (22 measurements)

22 measurements — page 2 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	45.0	51.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate , p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	56.0	63.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate , p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")

Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100.0	18.0	%	0.12 M Hepes buffer, 50 µlitre CoCl2	7.4	25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100.0	4.0	%	0.12 M Hepes buffer, 50 µlitre CoCl2	7.4	25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.0	%	0.12 M Hepes buffer, 50 µlitre CoCl2	7.4	25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100.0	0.0	%	0.12 M Hepes buffer, 50 µlitre CoCl2	7.4	25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	42.0	%	0.12 M Hepes buffer, 50 µlitre CoCl2	7.4	25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in %)
Stability - Half-life	Enzyme half-life (in days), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm)	Dimethyl Sulfoxide (DMSO)	20%	1.5	1.0	hours	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 50°C, Assay: 25°C	? mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in hours)
Stability - Half-life	Enzyme half-life (in days), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm)	Dimethyl Sulfoxide (DMSO)	20%	1.8	7.1	hours	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	? mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Classical aqueous control (in hours)
Stability - Incubation	Activity after incubation (3 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	33.0	50.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	56.0	89.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	45.0	69.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (3 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	33.0	64.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	56.0	85.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	45.0	61.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (3 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	33.0	59.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	56.0	96.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	45.0	71.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (3 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	33.0	62.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	60%	56.0	39.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	60%	45.0	34.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (3 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	60%	33.0	29.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (2 days at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	45.0	51.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")
Stability - Incubation	Activity after incubation (1 day at 25°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 412 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	56.0	63.0	%	Incubation: 0.12 M Hepes buffer, 50 µM CoCl2, Assay: 0.12 M Hepes buffer, 50 µM CoCl2	Incubation: 7.4, Assay: 7.4	Incubation: 25°C, Assay: 25°C	0.5 mM Paraoxon	diethyl phosphate, p-Nitrophenol	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous phase ("assayed with 0.5 mM substrate in buffer with less than 1% DMSO")

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Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability — Half-life

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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Phosphotriesterase from Pseudomonas diminuta

Enzyme Description

Sequence

Experimental Data (22 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Visualization : Stability — Half-life

Structure

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