Phospholipase A1 from Serratia sp. MK1

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 320 amino acids
MSMSLSFTSAIAPAAIQPPMVRTQPEPLSSSQPVEASATKAPVATLSQNSLNAQSLLNTLVSEISAAAPAAANQGVTRGQQPQKGDYTLALLAKDVYSTGSQGVEGFNRLSADALLGAGIDPASLQDAASGFQAGIYTDNQQYVLAFAGTNDMRDWLSNVRQATGYDDVQYNQAVSLAKSAKAAFGDALVIAGHSLGGGLAATAALATGTVAVTFNAAGVSDYTLNRMGIDPAAAKQDAQAGGIRRYSEQYDMLTGTQESTSLIPDAIGHKITLANNDTLSGIDDWRPSKHLDRSLTAHGIDKVISSMAEQKPWEAMANA
Jae Kwang Song et al. (2001) β€” Enhancement of stability and activity of phospholipase A1 in organic solvents by directed evolution
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology  Β· doi:10.1016/s0167-4838(01)00204-7 β†—  Β· Activity - Classical Stability - Incubation
68 measurements
Database ID
UniProt: Q9X9D8 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli XL1-Blue

Experimental Data (68 measurements)

68 measurements β€” page 1 of 4
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
WT Activity - Classical Activity measured by TLC-FID in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 β€” 56 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
WT Activity - Classical Activity measured by TLC-FID in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 β€” 27 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
WT Activity - Classical Activity measured by TLC-FID in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 β€” 13 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
WT Activity - Classical Activity measured by TLC-FID in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100 β€” 5 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
WT Activity - Classical Activity measured by TLC-FID in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50% 100 β€” 3 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100 β€” 19 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase n-Octane 100% 100 β€” 18 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase n-Hexane 100% 100 β€” 13 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Toluene 100% 100 β€” 19 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Butyl Acetate 100% 100 β€” 21 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Acetonitrile 100% 100 β€” 6 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethylformamide (DMF) 100% 100 β€” 2 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 100% 100 β€” 2 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 30% 100 β€” 5 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 100 β€” 3 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 50% 100 β€” 2 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" assay Non-incubated control (in %), assay in aqueous phase
WT Stability - Incubation Activity after incubation (10 hours at 37Β°C), measured by measuring product concentration by TLC-FID in aqueous phase Dimethyl Sulfoxide (DMSO) 10% 100 β€” 50 % Incubation: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS, Assay: 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS Incubation: 9, Assay: 9 Incubation: 37Β°C, Assay: 40Β°C 2 mL of 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” 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 3 57 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” 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 3 23 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” 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 5 38 % 50 mM Tris-HCl, 10 mM CaCl2, 450 Β΅M CHAPS 9 40Β°C 20 Β΅M 1,2-dimyristoyl-sn-glycero-3-phosphocholine 2-myristoyl-lysophosphatidylcholine , Myristic acid β€” Yes, "thorough shaking" Classical aqueous control (in %)
β€Ή Prev
1 2 3 4

Mutations in this dataset (4)

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

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” 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 β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” 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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