Phospholipase A1 from Serratia sp. MK1

Enzyme Description

Species

Extremophile

EC Number

3.1.1.32 ↗

Sequence

Length: 320 amino acids

MSMSLSFTSAIAPAAIQPPMVRTQPEPLSSSQPVEASATKAPVATLSQNSLNAQSLLNTLVSEISAAAPAAANQGVTRGQQPQKGDYTLALLAKDVYSTGSQGVEGFNRLSADALLGAGIDPASLQDAASGFQAGIYTDNQQYVLAFAGTNDMRDWLSNVRQATGYDDVQYNQAVSLAKSAKAAFGDALVIAGHSLGGGLAATAALATGTVAVTFNAAGVSDYTLNRMGIDPAAAKQDAQAGGIRRYSEQYDMLTGTQESTSLIPDAIGHKITLANNDTLSGIDDWRPSKHLDRSLTAHGIDKVISSMAEQKPWEAMANA

Jae Kwang Song et al. (2001) — Enhancement of stability and activity of phospholipase A1 in organic solvents by directed evolution

Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology · doi:10.1016/s0167-4838(01)00204-7 ↗ · Activity - Classical Stability - Incubation

68 measurements ▼

Database ID

UniProt: Q9X9D8 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli XL1-Blue

Experimental Data (68 measurements)

68 measurements — page 2 of 4

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
V34A/L292Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100	13	48	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
V34A/L292Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100	27	54	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
V34A/L292Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100	56	74	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
D168E/A234S/A235V/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100	27	76	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
D168E/A234S/A235V/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100	5	60	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
D168E/A234S/A235V/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100	13	70	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
Y251F/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100	56	83	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
Y251F/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100	27	68	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
Y251F/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100	13	60	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
Y251F/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100	5	52	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
Y251F/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100	3	36	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
D168E/A234S/A235V/L263Q	Activity - Classical	Activity measured by TLC-FID in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100	56	94	%	50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	9	40°C	20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking"	Classical aqueous control (in %)
D168E/A234S/A235V/L263Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	100	3	39	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
Y251F/L263Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethylformamide (DMF)	100%	100	2	15	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
Y251F/L263Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	100%	100	2	27	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
V34A/L292Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	50%	100	2	25	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
V34A/L292Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	100	3	29	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
V34A/L292Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	100	5	39	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
V34A/L292Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	100	19	52	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase
V34A/L292Q	Stability - Incubation	Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100	50	78	%	Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS	Incubation: 9, Assay: 9	Incubation: 37°C, Assay: 40°C	2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine	2-myristoyl-lysophosphatidylcholine , Myristic acid	—	Yes, "thorough shaking" assay	Non-incubated control (in %), assay in aqueous phase

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10%

100

—

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20%

100

—

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30%

100

—

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

40%

100

—

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

—

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

20%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Octane

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Hexane

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Toluene

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Butyl Acetate

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Acetonitrile

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethylformamide (DMF)

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

100%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

30%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

40%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

100

—

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

V34A/L292Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

V34A/L292Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

40%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

V34A/L292Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

V34A/L292Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

V34A/L292Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

D168E/A234S/A235V/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

D168E/A234S/A235V/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

40%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

D168E/A234S/A235V/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Y251F/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Y251F/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Y251F/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Y251F/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

40%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

Y251F/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

D168E/A234S/A235V/L263Q

Activity - Classical

Activity measured by TLC-FID in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10%

100

50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

40°C

20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking"

Classical aqueous control (in %)

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

40%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethylformamide (DMF)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

40%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

30%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

20%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Acetonitrile

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

30%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

20%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

40%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

30%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

20%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Octane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethyl Sulfoxide (DMSO)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethylformamide (DMF)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Acetonitrile

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Butyl Acetate

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Toluene

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Hexane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

D168E/A234S/A235V/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Octane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Hexane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Dimethylformamide (DMF)

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Acetonitrile

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Butyl Acetate

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Toluene

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Hexane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

V34A/L292Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

n-Octane

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Toluene

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

Y251F/L263Q

Stability - Incubation

Activity after incubation (10 hours at 37°C), measured by measuring product concentration by TLC-FID in aqueous phase

Butyl Acetate

100%

100

Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 µM CHAPS

Incubation: 9, Assay: 9

Incubation: 37°C, Assay: 40°C

2 mL of 20 µM 1,2-dimyristoyl-sn-glycero-3-phosphocholine

2-myristoyl-lysophosphatidylcholine, Myristic acid

—

Yes, "thorough shaking" assay

Non-incubated control (in %), assay in aqueous phase

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Mutations in this dataset (4)

WT V34A/L292Q D168E/A234S/A235V/L263Q Y251F/L263Q

Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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Phospholipase A1 from Serratia sp. MK1

Enzyme Description

Sequence

Experimental Data (68 measurements)

Visualization : Activity — Classical

Mutation Effect

Visualization : Stability — Incubation

Mutation Effect

Structure

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