Lipase (gene LipAMS8) from Pseudomonas sp. AMS8

Enzyme Description

Species

Pseudomonas sp. AMS8 ↗

Extremophile

Yes psychrophilic organism

EC Number

3.1.1.3 ↗

Sequence

Length: 476 amino acids

MGVFDYKNLGTEGSKALFADAMAITLYSYHNLDNGFAVGYQHNGFGLGLPATLVGALLGSTDSQGVIPGIPWNPDSEKAALEAVNKAGWTPISASTLGYGGKVDARGTFFGEKAGYTTAQVEVLGKYDGDGKLLEIGIGFRGTSGPRETLITDSIGDLVSDLLAALGPKDYAKNYAGEAFGTLLKDVAAYAGSHGLTGKDVVVSGHSLGGLAVNSMADLSGNKWSGFYKDSNYVAYASPTQSSGDKVLNIGYENDPVFRALDGSSFNFSSLGVHDKPHESTTDNIVSFNDHYASTLWNVLPFSIVNVPTWLSHLPTGYGDGLTRVLDSKFYDLTSRDSTIIVANLSDPARANTWVQDLNRNAEPHKGNTFIIGSDGNDLIQGGKGVDFIEGGKGNDTIRDNSGHNTFLFGGQFGQDRVIGYQSTDKLVFKDVEGSADWRDHAKVVGGDTVLSFGADSVTLVGVGLAGVGGDGISIS

Menega Ganasen et al. (2016) — Cold-adapted organic solvent tolerant alkalophilic family I.3 lipase from an Antarctic Pseudomonas

International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2016.06.095 ↗ · Activity - Michaelis-Menten Stability - Incubation

43 measurements ▼

Database ID

UniProt: E1B2U7 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (43 measurements)

43 measurements — page 3 of 3

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 (30 minutes at 15°C), measured by absorbance spectrophotometry (Kwon and Rhee colorimetric assay, absorbance of copper acetate–pyridine and free fatty acids reaction product measurement, 715 nm) in aqueous phase	n-Tridecane	25%	100.0	135.5	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 7, Assay: 7	Incubation: 15°C, Assay: 30°C	35% Olive oil	free fatty acids , glycerol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (30 minutes at 15°C), measured by absorbance spectrophotometry (Kwon and Rhee colorimetric assay, absorbance of copper acetate–pyridine and free fatty acids reaction product measurement, 715 nm) in aqueous phase	n-Tetradecane	25%	100.0	117.3	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 7, Assay: 7	Incubation: 15°C, Assay: 30°C	35% Olive oil	free fatty acids , glycerol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (30 minutes at 15°C), measured by absorbance spectrophotometry (Kwon and Rhee colorimetric assay, absorbance of copper acetate–pyridine and free fatty acids reaction product measurement, 715 nm) in aqueous phase	n-Hexadecane	25%	100.0	113.6	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 7, Assay: 7	Incubation: 15°C, Assay: 30°C	35% Olive oil	free fatty acids , glycerol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Methanol

135.8

115.5

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Isopropanol

5.1

37.2

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Toluene

5.1

2.9

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

n-Hexane

5.1

4.7

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

216.5

286.4

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Methanol

216.5

189.5

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Ethanol

216.5

650.6

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Isopropanol

216.5

573.2

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Toluene

216.5

194.6

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

Km (in nm) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

n-Hexane

216.5

278.2

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nm)

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

135.8

172.9

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Ethanol

5.1

47.4

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Ethanol

135.8

738.9

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Isopropanol

135.8

541.7

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Toluene

135.8

84.1

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

kcat (in 1/s) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

n-Hexane

135.8

100.2

1/s

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in 1/s)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

0.6

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Methanol

0.6

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Ethanol

0.6

1.1

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Isopropanol

0.6

0.9

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Toluene

0.6

0.4

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

kcat/kM (nM^-1 * s^-1) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

n-Hexane

0.6

0.3

nM^-1 * s^-1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in nM^-1 * s^-1)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Methanol

5.1

5.2

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Activity - Michaelis-Menten

Vmax (V) determined by p-nitrophenol absorbance measurement (405 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

5.1

9.9

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

25°C

p-Nitrophenyl palmitate

p-Nitrophenol, Palmitic Acid

—

Classical aqueous control (in V)

Stability - Incubation

Activity after incubation (30 minutes at 15°C), measured by absorbance spectrophotometry (Kwon and Rhee colorimetric assay, absorbance of copper acetate–pyridine and free fatty acids reaction product measurement, 715 nm) in aqueous phase

Ethanol

25%

100.0

88.3

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

25%

100.0

122.3

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Methanol

25%

100.0

101.9

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Pyridine

25%

100.0

0.0

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

1-Butanol

25%

100.0

0.2

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Isoamyl Alcohol

25%

100.0

0.9

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Propyl Acetate

25%

100.0

7.0

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

1-Propanol

25%

100.0

4.7

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Benzene

25%

100.0

66.1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Toluene

25%

100.0

78.1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Xylene

25%

100.0

91.9

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Hexane

25%

100.0

101.4

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

1-Decanol

25%

100.0

24.4

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

Isooctane

25%

100.0

118.4

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Octane

25%

100.0

108.8

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Decane

25%

100.0

126.1

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Tridecane

25%

100.0

135.5

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Tetradecane

25%

100.0

117.3

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Stability - Incubation

n-Hexadecane

25%

100.0

113.6

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 7, Assay: 7

Incubation: 15°C, Assay: 30°C

35% Olive oil

free fatty acids, glycerol

—

Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

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Visualization : Activity — Michaelis-Menten

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

Visualization : Stability — Incubation

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

Structure

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Lipase (gene LipAMS8) from Pseudomonas sp. AMS8

Enzyme Description

Sequence

Experimental Data (43 measurements)

Visualization : Activity — Michaelis-Menten

Visualization : Stability — Incubation

Structure

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