Information on EC 3.1.1.3 - triacylglycerol lipase and Organism(s) Pseudomonas aeruginosa and UniProt Accession P26876

-

665475, 682482

alkaline lipase

-

amano AP

-

amano B

-

amano CE

-

amano CES

-

amano P

-

amno N-AP

-

BAL

-

Bile-salt-stimulated lipase

-

BSSL

-

butyrinase

-

cacordase

-

CALB

-

capalase L

-

Carboxyl ester lipase

-

cholesterol esterase

-

CS-2 lipase

-

Cytotoxic T lymphocyte lipase

-

EDL

-

enantiospecific lipase

-

endothelial cell-derived lipase

-

endothelial-derived lipase

-

extracellular lipase

-

664156, 665893, 666250, 682488

GA 56 (enzyme)

-

Gastric lipase

-

GEH

-

glycerol ester hydrolase

glycerol-ester hydrolase

-

heparin releasable hepatic lipase

-

hepatic lipase

-

hepatic monoacylglycerol acyltransferase

-

Lingual lipase

-

Lip9

-

LipA

-

lipase

lipase I

-

lipase II

-

lipase, triacylglycerol

-

lipazin

-

LipH

-

liver lipase

-

LST-03 lipase

-

meito MY 30

-

meito Sangyo OF lipase

-

organic solvent-stable lipase

-

Pancreatic lipase

-

Pancreatic lysophospholipase

-

PGE

-

PL-RP2

-

post-heparin plasma protamine-resistant lipase

-

PPL

-

Pregastric esterase

-

Pregastric lipase

-

PseA

-

salt-resistant post-heparin lipase

-

steapsin

-

Sterol esterase

-

takedo 1969-4-9

-

teenesterase

-

tiacetinase

-

tibutyrin esterase

-

triacylglycerol acylhydrolase

-

triacylglycerol ester hydrolase

-

Triacylglycerol lipase

-

tributyrase

-

tributyrinase

-

triglyceridase

-

triglyceride hydrolase

-

triglyceride lipase

-

triolein hydrolase

-

tween hydrolase

-

tween-hydrolyzing esterase

-

Tweenase

-

additional information

the enzyme belongs to the family 1.1 of bacterial lipases

triacylglycerol + H2O = diacylglycerol + a carboxylate

the catalytic triad is formed by Ser82, Asp229, and His251, substrates bind first to Ser82, position of the oxyanion hole and the three pockets accomodating the sn-1, sn-2, and sn-3 fatty acid chains, reaction mechanism, tetrahedral intermediate during acylation step

triacylglycerol + H2O = diacylglycerol + a carboxylate

hydrolysis of carboxylic ester

-

BRENDA

lipid metabolism

KEGG

Glycerolipid metabolism

-

-, -

triacylglycerol acylhydrolase

The enzyme is found in diverse organisms including animals, plants, fungi, and bacteria. It hydrolyses triglycerides into diglycerides and subsequently into monoglycerides and free fatty acids. The enzyme is highly soluble in water and acts at the surface of oil droplets. Access to the active site is controlled by the opening of a lid, which, when closed, hides the hydrophobic surface that surrounds the active site. The lid opens when the enzyme contacts an oil-water interface (interfacial activation). The pancreatic enzyme requires a protein cofactor, namely colipase, to counteract the inhibitory effects of bile salts.

9001-62-1

-

triacylglycerol + H2O

diacylglycerol + a carboxylate

-

-

?

1,2-dilauryl-rac-glycero-3-glutaric acid resorufinester + H2O

?

-

246540

-

?

2,3-dimercaptopropan-1-ol tributyrate + H2O

?

2-methyldecanoic acid 4-nitrophenyl ester + H2O

(S)-2-methyldecanoate + 4-nitrophenol + (R)-2-methyldecanoic acid 4-nitrophenyl ester

-

model substrate, enantioselectivity in the asymmetric hydrolysis with preference for the S-enantiomer

-

?

3 triolein + 3 H2O

1,3-diolein + 1,2-diolein + 2,3-diolein + 3 oleate

-

shows random positional specificity for triolein hydrolysis

-

?

4-nitrophenyl 2-methyldecanoate + H2O

?

-

kinetic resolution of enantioselectivity, overview

-

?

4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate + H2O

?

-

reaction via R-tetrahedral intermediate and S-tetrahedral intermediate, overview, the wild-type lipase and the mutant L162F show preference for the (+)-enantiomer, the mutant shows high enantioselectivity, kinetic resolution of enantioselectivity, overview

-

?

4-nitrophenyl acetate + H2O

4-nitrophenol + acetate

-

-

?

4-nitrophenyl butyrate + H2O

4-nitrophenol + butyrate

-

-

?

4-nitrophenyl caprate + H2O

4-nitrophenol + caprate

-

-

?

4-nitrophenyl laurate + H2O

4-nitrophenol + laurate

4-nitrophenyl myristate + H2O

4-nitrophenol + myristate

4-nitrophenyl palmitate + H2O

4-nitrophenol + palmitate

4-nitrophenyl palmitate + H2O

palmitate + 4-nitrophenol

-

the immobilized enzyme PAL PCMC works as better catalyst for hydrolysis of pNPP in n-heptane medium

-

?

4-nitrophenyl stearate + H2O

4-nitrophenol + stearate

-

-

?

acylglycerol + H2O

glycerol + a carboxylate

-

-

?

butyl butyrate + H2O

butane + butyrate

-

-

?

castor oil + H2O

?

-

-

?

diacylglycerol + H2O

acylglycerol + a carboxylate

-

-

?

ethyl butyrate + H2O

ethane + butyrate

-

-

?

methyl oleate + H2O

methanol + oleate

-

acylglycerol lipase activity, EC 3.1.1.23

-

?

oleoyl 2-naphthyl ester + H2O

oleic acid + 2-naphthol

-

-

?

oleoyl 2-naphthylamide + H2O

oleic acid + 2-naphthylamine

-

-

?

soybean oil + H2O

?

-

concentrated soybean oil, high activity in a water-restricted environment containing a water content of 0.5-1% w/w

-

?

trans-3-(4-methoxyphenyl) glycidic acid methyl ester + H2O

?

-

i.e. (+-)-methyl trans-3(4-methoxyphenyl) glycidate, the substrate is a key intermediate in the synthesis of cardiovascular drug, diltiazem

-

?

triacetin + H2O

diacetin + acetate

-

-

?

triacylglycerol + H2O

diacylglycerol + a carboxylate

tributyrin + H2O

dibutyrin + butyrate

tricaprin + H2O

dicaprin + caprate

-

high activity

-

?

tricaproin + H2O

dicaproin + caproate

-

high activity

-

?

tricaprylin + H2O

dicaprylin + caprylate

-

best substrate

-

?

trimyristin + H2O

dimyristin + myristate

-

665891

-

?

triolein + H2O

diolein + oleate

trioleoylglycerol + H2O

oleic acid + ?

-

-

?

tripropionin + H2O

dipropionin + propionate

-

-

?

additional information

?

-

8 entries

triacylglycerol + H2O

diacylglycerol + a carboxylate

-

680948, 690498

-

?

additional information

?

-

Ca2+

stabilizes the loop containing the catalytic His251 residue, binding site structure

Ca2+

CaCl2

-

1.24fold stimulation at 1 mM

Mg2+

-

activates extracellular isozyme 2 slightly

Na+

-

activates extracellular isozyme1 slightly

taurocholic acid

-

1.6fold stimulation at 200 mM

Zn2+

-

activates extracellular isozyme 1 slightly, inhibits extracellular isozyme 2

additional information

-

the extracellular isozymes are not affected by SDS

Rc-(Rp,Sp)-1,2-dioctylcarbamoyl-glycero-3-O-(4-nitrophenyl) octylphosphonate

binding site structure, binding arrests the enzyme in the open conformation

2-mercaptoethanol

-

reduces the enzyme activity to 48% after 1 h

3,4-dichloroisocoumarin

-

weak inhibition

acetonitrile

-

Ca2+

Cd2+

-

strain KKA-5, slight inhibition

cetyltrimethylammonium bromide

-

45% residual activity after 5 h at 25% (v/v)

CHAPS

-

86% residual activity after 5 h at 25% (v/v)

Cu2+

diethyl 4-nitrophenyl phosphate

-

rapid and complete inhibition

diisopropyl fluorophosphate

-

30% inhibition at 5 mM, 60% inhibition at 25 mM

diisopropylfluorophosphate

-

82% residual activity after 30 min at 8 mM

DTT

-

EDTA

eserine

-

27% inhibition at 5 mM

Fe2+

-

strain KKA-5, strong inhibition

Fe3+

-

strain KKA-5, strong inhibition

Hg2+

hydrogen peroxide

-

62% residual activity after 5 h at 25% (v/v)

Mg2+

-

32% inhibition at 5 mM

Mn2+

p-chloromercuric benzoate

-

20% residual activity after 30 min at 8 mM

PMSF

protamine

-

50% inhibition at 2 mg/ml

SDS

sodium deoxycholate

-

90% residual activity after 5 h at 25% (v/v)

sodium hypochlorite

-

85% residual activity after 5 h at 25% (v/v)

sodium taurocholate

-

67% inhibition at 5 mM

Triton X-100

-

Tween 20

-

74% residual activity after 5 h at 25% (v/v)

Zn2+

4 entries

additional information

8 entries

-

castor oil

-

at 2% as carbon source, enhances the enzyme production in vivo, pH 6.9

chloroform

-

at 25% v/v, 30°C, 130% activity after 24 h, 140% after 48 h

Dimethyl formamide

-

concentrated solution increases the enzyme activity 2fold, maximal activity at a water content of 0.5-1% w/w

dimethyl sulfoxide

-

concentrated solution increases the enzyme activity 5fold, maximal activity at a water content of 0.5-1% w/w

dimethylformamide

-

at 25% v/v, 30°C, 120% activity after 24 h, 110% after 48 h

DMSO

-

activates both extracellular isozymes 1.73fold

DTT

-

20% activation at 5 mM

EDTA

-

30% activation at 5 mM

Fe3+

-

132% relative activity after 30 min at 1 mM

glutathione

-

112% relative activity after 30 min at 1 mM

iodoacetic acid

-

10% activation at 5 mM

sodium deoxycholate

-

300% activation at 5 mM

Triton X-100

Tween 80

additional information

-

50 - 679.8

4-nitrophenyl palmitate

4 entries

3000

triolein

-

additional information

-

-

0.00006

-

purified enzyme, 95% acetone, 30°C

0.00012

-

purified enzyme, 95% ethyl-methyl-ketone, 30°C

0.00013

-

purified enzyme, 95% n-hexane, 30°C

0.0052

-

purified enzyme, 95% ethanol, 30°C

0.231

-

purified enzyme, 95% methanol, 30°C

143.3

-

after 8.6fold purification, in Tris-HCl, buffer (0.1 M, pH 8.0)

16.6

-

crude filtrate, in Tris-HCl, buffer (0.1 M, pH 8.0)

165.5

-

purified extracellular isozyme 1

19280

-

purified enzyme

22.22

-

purified enzyme

384

-

purified extracellular isozyme 2

473

-

purified enzyme, buffer

additional information

10

-

extracellular isozyme 1

11

-

6

-

assay at

6 - 7

-

7

7.5

-

assay at

690408

8

4 entries

8.5

-

strain KKA-5

8.9

-

extracellular enzyme

9

-

extracellular isozyme 2

4 - 11.5

-

activity range

4 - 8

-

5 - 11

-

5.5 - 9

-

6 - 10

-

20

-

assay at

30

35

-

40

45

50

-

extracellular isozyme 2

55

-

strain MB5001

70

-

25 - 50

-

25 - 90

-

30 - 50

almost no activity above 50°C

30 - 90

-

4.9

-

5.5

-

isoelectric focusing

5.7

-

isoelectric focusing

7.3 - 7.4

-

strain F-111

-

665475, 682482

SwissProt

culture medium

-

the enzyme is excreted during the late logarythmic growth phase

additional information

1ex9, download, 3D-view

SCOPe (1ex9)

CATH (1ex9)

29000

x * 29000, crystal structure

682482

15500

-

1 * 15500, extracellular isozyme 1, SDS-PAGE, 1 * 54970, extracellular isozyme 2, SDS-PAGE

280000

-

peak II, gel filtration

29000

30000

-

x * 29000, enzyme of strain MB5001, SDS-PAGE, x * 32000, enzyme from strain F-111, SDS-PAGE, x * 30000, enzyme of strain KKA-5, SDS-PAGE

300000

-

gel filtration

32000

-

x * 29000, enzyme of strain MB5001, SDS-PAGE, x * 32000, enzyme from strain F-111, SDS-PAGE, x * 30000, enzyme of strain KKA-5, SDS-PAGE

35700

-

x * 35700, estimated from SDS-PAGE

40000

-

x * 40000, about, SDS-PAGE

54000

-

x * 54000, SDS-PAGE

54970

-

1 * 15500, extracellular isozyme 1, SDS-PAGE, 1 * 54970, extracellular isozyme 2, SDS-PAGE

59400

-

x * 59400, SDS-PAGE

60000

-

SDS-PAGE

840000

-

peak I, gel filtration

additional information

-

the enzyme tends to form aggregates

?

x * 29000, crystal structure

?

monomer

oligomer

additional information

6 entries

proteolytic modification

-

protease V8 cleaves between surface residues Asp38 and Glu46

purified recombinant enzyme in open conformation through complexion by inhibitor Rc-(Rp,Sp)-1,2-dioctylcarbamoyl-glycero-3-O-(4-nitrophenyl) octylphosphonate, sitting drop vapour diffusion method, room temperature, 5 mg/ml protein in10 mM Tris-HCl, pH 8.0, 1% beta-octylpyranoside, equilibration against precipitation solution containing 25% 2-methyl-2,4-pentanediol, 20 mM CaCl2, 100 mM citrate, pH 5.6, several months, X-ray structure determination and analysis at 2.54 A resolution, structure scheme and modelling

A217S

the mutant movement of the lids is very similar to that in the wild-type lipase

682482

A213D

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

A213D/F265L

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

D20N/S53P/S155M/L162G/T180I/T234S

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

F207S

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

F207S/A213D

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

F207S/A213D/F265L

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

F207S/F265L

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

F265L

-

random mutagenesis, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

L159W/L162T

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162A

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162D

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162F

-

site-directed mutagenesis, a highly enantioselective mutant, use in combinatorial active site saturation test for kinetic resolution of an axially chiral allene, 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate, the high enantioselectivity of the mutant is rationalized by PIPI stacking

L162G

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162G/L159V

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162G/L159Y

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162I

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162T

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L162V

-

site-directed mutagenesis, the mutant shows higher enantioselectivity as the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L17F

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

L231e/V232C

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

M16A

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

P96H/F207S

-

random mutagenesis, double mutation plus an additional silent mutation, the mutation occurs at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site, and leads to increased amidase activity of the enzyme

S53P/L162G

-

optimizing enantioselectivity of LipA by comprehensive directed evolution approach

T180I/T234S

-

identification of a mutant LipH 1H8 with increased (S)-enantioselectivity for model substrate 2-methyldecanoic acid 4-nitrophenyl ester compared to the wild-type enzyme, overview. The mutant shows low secretion efficiency, identification of two amino acid substitutions located on the protein surface, which significantly impair lipase secretion, the amino acid substitutions T180I and T234S of lipase variant 1H8 are located in close vicinity to these cysteines, suggesting that these substitutions may impair its ability to form disulfide bonds

V232I

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

V232I/M16L/A34T/P86L/D113G/S237T/T150A/S147N/V94A/T87S/L208H

-

site-directed mutagenesis, the mutant shows reduced enantioselectivity compared to the wild-type enzyme with substrates 4-nitrophenyl 4-cyclohexyl-2-methylbuta-2,3-dienoate and 4-nitrophenyl 2-methyldecanoate

additional information

10

-

30°C, 3 h, purified enzyme, loss of 50% activity

682607

4 - 11.5

-

stable

680948

5 - 8

stable

666368

6

-

30°C, 48 h, purified enzyme, loss of 33% activity

682607

6 - 8.5

-

694844

7 - 9

-

30°C, 48 h, purified enzyme, completely stable

682607

100

-

inactivation

20 - 55

-

purified enzyme, stable for over 30 days in presence of water-miscible organic solvents such as alcohols, glycols, pyridine, acetonitrile, dimethyl formamide, or dimethyl sulfoxide

25 - 50

-

37

stable up to

45

inactivation above

50

-

50 min, 50% remaining activity

60

70

80

-

complete inactivation within 10 min

additional information

lyophilization of the crude enzyme leads to only slight loss of activity

-

purified recombinant enzyme, 30°C, pH 8.0, half-life is 12.3 days

stable only in presence of detergents

-

the purified enzyme is very stable to freezing and thawing

-

the synthesis of butyl acetate in heptane decreases from 98.2% to 87.4% after 5 cycles in reuse of the immobilized lipase

-

1,4-butanediol

half-life is 7.4 days

1,5-pentanediol

half-life is 0.19 days

Acetone

-

at 25% v/v, 30°C, 95% remaining activity after 24 h, 33% after 48 h

Brij 35

-

121% relative activity after 5 h at 25% (v/v)

Butanol

-

at 25% v/v, 30°C, 10% remaining activity after 48 h

chloroform

-

at 25% v/v, 30°C, 130% activity after 24 h, 140% after 48 h

cyclohexane

half-life is 0.48 days

dimethyl formamide

-

the purified enzyme is stable for over 30 days at 20-55°C in presence of water-miscible organic solvents such as alcohols, glycols, pyridine, acetonitrile, dimethyl formamide, or dimethyl sulfoxide

dimethyl sulfoxide

dimethylformamide

-

at 25% v/v, 30°C, 120% activity after 24 h, 110% after 48 h, 100% remaining activity after 30 min at 50°C

Ethanol

ethyl methyl ketone

-

75% remaining activity after 10 min at 50°C

Ethylene glycol

half-life is 84.6 days

Glycerol

half-life is 37 days

heptane

-

the recombinant lipase shows stability in the presence of heptane

hexane

-

at 25% v/v, 30°C, 100% remaining activity after 24 h, 64% after 48 h, 100% reaming activity after 15 min at 60°C, 50% reamining activity after 30 min at 60°C

isopropanol

-

at 25% v/v, 30°C, 65% remaining activity after 24 h, 16% after 48 h

Methanol

N,N-dimethylformamide

half-life is 7.9 days

n-decane

half-life is 0.40 days

n-heptane

half-life is 7.9 days

n-hexane

half-life is 7.7 days

n-octane

half-life is 9.2 days

Pyridine

-

the purified enzyme is stable for over 30 days at 20-55°C in presence of water-miscible organic solvents such as alcohols, glycols, pyridine, acetonitrile, dimethyl formamide, or dimethylsulfoxide

tert-Butanol

half-life is 0.17 days

toluene

half-life is 0.37 days

Tween 80

-

124% relative activity after 5 h at 25% (v/v)

additional information

5 entries

-20°C, crude enzyme, stable for at least one month with repeated freezing and thawing

-

22°C, room temperature, the crude enzyme is stable for a few hours

-

4°C, overnight, loss of 60% activity

-

purified recombinant enzyme, 30°C, pH 8.0, half-life is 12.3 days

-

-

80833, 246540

extracellular enzyme 35fold from culture medium by gel filtration and disc electrophoresis

-

extracellular enzyme by chromatographical isoelectric focusing

-

from medium by gel filtration

-

from medium of a Tween 80-limited culture, by anion exchange chromatography and gel filtration

-

from strain MB5001 in a 3-step procedure, from strain F-111 to homogeneity, 518fold from strain KKA-5 to homogeneity

-

method development for a modified blue native polyacrylamide gel electrophoresis protocol that can overcome aggregation of lipases seen in native PAGE and solve the functional enzyme, overview

-

690408

native enzyme 12.5fold from culture broth by ammonium sulfate fractionation, hydrophobic interaction chromatography, dialysis, and gel filtration to homogeneity

-

native extracellular enzyme 21fold from the culture supernatant by acetone precipitation, anion exchange chromatography and gel filtration

-

native extracellular enzyme from culture supernatant by acetone precipitation and hydrophobic interaction chromatography

-

nickel affinity column chromatography

-

recombinant enzyme from Escherichia coli strain JM109 by acetone fractionation, ion exchange and hydrophobic interaction chromatography to homogeneity

recombinant enzyme, after solubilization from inclusion bodies, by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, and gel filtration to homogeneity

recombinant wild-type LipA and LipH mutant 1H8 from Escherichia coli strain BL21(DE3) solubilized inclusion bodies by

-

two native extracellular isozymes from strain Ps-x by ammonium sulfate fractionation, and anion and cation exchange chromatography, extracellular isozyme 1 is purified 6.36fold, and extracellular isozyme 2 is purified 14.76fold

-

ultrafiltration and Sephadex G-100 gel filtration

-

co-expression of CS-2 lipase and its cognate foldase in Escherichia coli BL21(DE3) cells

-

gene lip, chromosomal mapping, DNA sequence determination and analysis, expression in Escherichia coli strains SK 1108 and JM109, the latter deficient in extracellular lipase

-

665891

gene lip3, shotgun cloning, construction of a genomic library, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain JM109

gene lip9, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain BL21(DE3) inclusion bodies, subcloning in Escherichia coli strain JM109

overexpression of wild-type enzymes, and mutant LipA and LipH mutant 1H8 in Escherichia coli strain BL21(DE3) in inclusion bodies, subcloning in Escherichia coli strain DH5alpha, expression of mutant enzymes in Pseudomonase aeruginosa strains PABS1 and PABST7.1

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strain IGB83, cloning and functional expression in Xanthomonas campestris, secretion of the recombinant enzyme to the medium, expression of the enzyme also in Escherichia coli as His-tagged protein, secretion to the medium

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subcloning in Escherichia coli strains DH5alpha and JM109, expression of wild-type and mutant enzymes in Pseudomonas aeruginosa strain PAO1162

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recombinant enzyme from Escherichia coli inclusion bodies by 20 mM Tris-HCl buffer, pH 8.0, and 5.3 M urea, followed by a 100fold dilution

recombinant wild-type LipA and LipH mutant 1H8 from Escherichia coli strain BL21(DE3) inclusion bodies, denaturation by treatment for 1 h at 37°C with 100 mM Tris-HCl, pH 8.0, 8 M urea, followed by in vitro refolding to enzymatic activity performed for 3 h by adding equimolar ratios of LipH and sudden 10fold dilution in a buffer containing 50 mM Tris-HCl, pH 8.0, 3.5 mM CaCl2, 0.7 mM laurylmaltoside and 45% v/v glycerol

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biotechnology

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the alkaline lipase is stable and active in organic solvents and a water-restricted environment and has a great potential as a biotechnological tool e.g. in organosynthetic reactions in water-restricted medium or in control and prevention of metalworking fluid putrification in the metal industry

synthesis

Stuer, W.; Jaeger, K.E.; Winkler, U.K.

Purification of extracellular lipase from Pseudomonas aeruginosa

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Pseudomonas aeruginosa

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Substrate specificities of bacterial polyhydroxyalkanoate depolymerases and lipases: bacterial lipases hydolyze poly(omega-hydroxyalkanoates)

Appl. Environ. Microbiol.

61

3113-3118

1995

Bacillus subtilis, Burkholderia cepacia, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens

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Sharma, R.; Chisti, Y.; Banerjee, U.C.

Production, purification, characterization, and applications of lipases

Biotechnol. Adv.

19

627-662

2001

Acinetobacter calcoaceticus, Aspergillus niger, Aspergillus oryzae, Geobacillus stearothermophilus, Bacillus sp. (in: Bacteria), Burkholderia cepacia, Burkholderia sp., Moesziomyces antarcticus, Diutina rugosa, Rhizomucor miehei, Penicillium roqueforti, Hyphopichia burtonii, Proteus vulgaris, Pseudomonas sp., Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas oleovorans, Rhizopus arrhizus, Rhodotorula glutinis, Staphylococcus epidermidis, Penicillium wortmanii, Penicillium roqueforti IAM7268, Bacillus sp. (in: Bacteria) J33, Acinetobacter calcoaceticus BD 413, Geobacillus stearothermophilus L1, Pseudomonas sp. KM1-56

14550014

Shabtai, Y.; Daya-Mishne, N.

Production, purification, and properties of a lipase from a bacterium (Pseudomonas aeruginosa YS-7) capable of growing in water-restricted environments

Appl. Environ. Microbiol.

58

174-180

1992

Pseudomonas aeruginosa, Pseudomonas aeruginosa YS-7

1539972

Jaeger, K.E.; Adrian, F.J.; Meyer, H.E.; Hancock, R.E.W.; Winkler, U.K.

Extracellular lipase from Pseudomonas aeruginosa is an amphiphilic protein

Biochim. Biophys. Acta

1120

315-321

1992

Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC1R

1576157

Finkelstein, A.E.; Strawich, E.S.; Sonnino, S.

Characterization and partial purification of a lipase from Pseudomonas aeruginosa

Biochim. Biophys. Acta

206

380-391

1970

Pseudomonas aeruginosa, Pseudomonas aeruginosa ATCC 10145

4989950

Jaeger, K.E.; Ransac, S.; Koch, H.B.; Ferrato, F.; Dijkstra, B.W.

Topological characterization and modeling of the 3D structure of lipase from Pseudomonas aeruginosa

FEBS Lett.

332

143-149

1993

Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC1R

8405431

Nardini, M.; Lang, D.A.; Liebeton, K.; Jaeger, K.E.; Dijkstra, B.W.

Crystal structure of Pseudomonas aeruginosa lipase in the open conformation. The prototype for family 1.1 of bacterial lipases

J. Biol. Chem.

275

31219-31225

2000

Pseudomonas aeruginosa (P26876), Pseudomonas aeruginosa

10893416

665891

Wohlfarth, S.; Winkler, U.K.

Chromosomal mapping and cloning of the lipase gene of Pseudomonas aeruginosa

J. Gen. Microbiol.

134

433-440

1988

Pseudomonas aeruginosa, Pseudomonas aeruginosa PAO 2302

3139825

665892

Gilbert, E.J.; Drozd, J.W.; Jones, C.W.

Physiological regulation and optimization of lipase activity in Pseudomonas aeruginosa EF2

J. Gen. Microbiol.

137

2215-2221

1992

Pseudomonas aeruginosa, Pseudomonas aeruginosa EF2

1748874

Gilbert, E.J.; Cornish, A.; Jones, C.W.

Purification and properties of extracellular lipase from Pseudomonas aeruginosa EF2

J. Gen. Microbiol.

137

2223-2229

1991

Pseudomonas aeruginosa, Pseudomonas aeruginosa EF2

1748875

Jaeger, K.E.; Kharazmi, A.; Hoiby, N.

Extracellular lipase of Pseudomonas aeruginosa: biochemical characterization and effect on human neutrophil and monocyte function in vitro

Microb. Pathog.

10

173-182

1991

Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC 1R

1910141

Ogino, H.; Hiroshima, S.; Hirose, S.; Yasuda, M.; Ishimi, K.; Ishikawa, H.

Cloning, expression and characterization of a lipase gene (lip3) from Pseudomonas aeruginosa LST-03

Mol. Genet. Genomics

271

189-196

2004

Pseudomonas aeruginosa (Q9KJG6), Pseudomonas aeruginosa, Pseudomonas aeruginosa LST-03 (Q9KJG6), Pseudomonas aeruginosa LST-03

14740297

666381

Duong, F.; Soscia, C.; Lazdunski, A.; Murgier, M.

The Pseudomonas fluorescens lipase has a C-terminal secretion signal and is secreted by a three-component bacterial ABC-exporter system

Mol. Microbiol.

11

1117-1126

1994

Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas fluorescens B52

8022281

Carballeira, J.D.; Krumlinde, P.; Bocola, M.; Vogel, A.; Reetz, M.T.; Baeckvall, J.E.

Directed evolution and axial chirality: optimization of the enantioselectivity of Pseudomonas aeruginosa lipase towards the kinetic resolution of a racemic allene

Chem. Commun. (Camb. )

2007

1913-1915

2007

Pseudomonas aeruginosa

17695227

Ogino, H.; Katou, Y.; Akagi, R.; Mimitsuka, T.; Hiroshima, S.; Gemba, Y.; Doukyu, N.; Yasuda, M.; Ishimi, K.; Ishikawa, H.

Cloning and expression of gene, and activation of an organic solvent-stable lipase from Pseudomonas aeruginosa LST-03

Extremophiles

11

809-817

2007

Pseudomonas aeruginosa (A8QYB2), Pseudomonas aeruginosa LST-03 (A8QYB2), Pseudomonas aeruginosa LST-03

17657406

Karadzic, I.; Masui, A.; Zivkovic, L.I.; Fujiwara, N.

Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metalworking fluid

J. Biosci. Bioeng.

102

82-89

2006

Pseudomonas aeruginosa

17027868

682482

Cherukuvada, S.L.; Seshasayee, A.S.; Raghunathan, K.; Anishetty, S.; Pennathur, G.

Evidence of a double-lid movement in Pseudomonas aeruginosa lipase: insights from molecular dynamics simulations

PLoS Comput. Biol.

1

e28

2005

Pseudomonas aeruginosa (P26876), Pseudomonas aeruginosa

16110344

Saeed, H.M.; Zaghloul, T.I.; Khalil, A.I.; Abdelbaeth, M.T.

Purification and characterization of two extracellular lipases from Pseudomonas aeruginosa Ps-x

Pol. J. Microbiol.

54

233-240

2005

Pseudomonas aeruginosa, Pseudomonas aeruginosa Ps-x

16450840

Singh, S.; Banerjee, U.C.

Purification and characterization of trans-3-(4-methoxyphenyl) glycidic acid methyl ester hydrolyzing lipase from Pseudomonas aeruginosa

Process Biochem.

42

1063-1068

2007

Pseudomonas aeruginosa, Pseudomonas aeruginosa MTCC 5113

-

Fujii, R.; Nakagawa, Y.; Hiratake, J.; Sogabe, A.; Sakata, K.

Directed evolution of Pseudomonas aeruginosa lipase for improved amide-hydrolyzing activity

Protein Eng. Des. Sel.

18

93-101

2005

Pseudomonas aeruginosa

15788423

690408

Saminathan, M.; Muthukumaresan, K.T.; Rengarajan, S.; Muthukrishnan, N.; Gautam, P.

Blue native electrophoresis study on lipases

Anal. Biochem.

377

270-271

2008

Pseudomonas aeruginosa, Pseudomonas aeruginosa MTCC-2297

18381198

Gaur, R.; Gupta, G.N.; Vamsikrishnan, M.; Khare, S.K.

Protein-coated microcrystals of Pseudomonas aeruginosa PseA lipase

Appl. Biochem. Biotechnol.

151

160-166

2008

Pseudomonas aeruginosa

18690417

Hausmann, S.; Wilhelm, S.; Jaeger, K.E.; Rosenau, F.

Catalytic mechanism of C-di-GMP specific phosphodiesterase: a study of the EAL domain-containing RocR from Pseudomonas aeruginosa

FEMS Microbiol. Lett.

282

65-72

2008

Pseudomonas aeruginosa

18355276

Gaur, R.; Gupta, A.; Khare, S.K.

Purification and characterization of lipase from solvent tolerant Pseudomonas aeruginosa PseA

Proc. Biochem.

43

1040-1046

2008

Pseudomonas aeruginosa, Pseudomonas aeruginosa PseA

-