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Literature summary extracted from

  • Koschorreck, K.; Liu, D.; Kazenwadel, C.; Schmid, R.D.; Hauer, B.
    Heterologous expression, characterization and site-directed mutagenesis of cutinase CUTAB1 from Alternaria brassicicola (2010), Appl. Microbiol. Biotechnol., 87, 991-997.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.74 PCR product digested with EcoRI and NotI, and ligated in corresponding sites of the linearized pPICZalphaA vector. Transformation of Escherichia coli DH5alpha cells. Wild-type and mutants heterologously expressed in Pichia pastoris X-33 cells under the control of the methanol-inducible AOX1 promoter Alternaria brassicicola

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.74 A84F mutation in the small helical flap, significantly increases the activity towards longer chain substrates like 4-nitrophenyl palmitate Alternaria brassicicola
3.1.1.74 I183A mutation in the hydrophobic binding loop, drastically reduces the overall activity Alternaria brassicicola
3.1.1.74 L181A mutation in the hydrophobic binding loop, drastically reduces the overall activity Alternaria brassicicola
3.1.1.74 L80A mutation in the hydrophobic binding loop, drastically reduces the overall activity Alternaria brassicicola

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.74 20000
-
 1 * 24000, SDS-PAGE, glycosylated CUTAB1. 1 * 20000, SDS-PAGE, non-glycosylated CUTAB1 Alternaria brassicicola
3.1.1.74 24000
-
 1 * 24000, SDS-PAGE, glycosylated CUTAB1. 1 * 20000, SDS-PAGE, non-glycosylated CUTAB1 Alternaria brassicicola

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.74 Alternaria brassicicola P41744 DSM 62008
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.1.1.74 glycoprotein the glycosylated form of CUTAB1 has a MW of 24000 Da. The non-glycosylated form of CUTAB1 has a MW of 20000 Da Alternaria brassicicola

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.74 wild-type and mutants purified by ultrafiltration, wild-type further purified by hydrophobic interaction chromatography and ammonium sulfate precipitation, 1.6fold with a yield of 70% Alternaria brassicicola

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.74 2.77
-
 carrier bound CUTAB1 with tributyrin as substrate, in 40 ml of 50 mM potassium phosphate buffer, pH 5.5 containing 25% ethanol, at 8°C, for 40 H Alternaria brassicicola
3.1.1.74 10
-
 crude extract, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 17
-
 1.6fold purified enzyme, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 17
-
 wild-type, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 30
-
 wild-type, with tripalmitin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 41
-
 wild-type, with methyl acetate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 47
-
 wild-type, with tristearin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 66
-
 mutant A84F, with tripalmitin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 77
-
 mutant A84F, with tristearin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 125
-
 wild-type, with ethyl caprylate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 125
-
 wild-type, with methyl caproate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 129
-
 wild-type, with methyl caprylate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 145
-
 wild-type, with methyl myristate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 157
-
 wild-type, with methyl propionate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 161
-
 wild-type, with methyl decanoate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 188
-
 mutant A84F, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 197
-
 wild-type, with methyl laurate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 257
-
 wild-type, with methyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 281
-
 wild-type, with ethyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 454
-
 wild-type, with triolein as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 586
-
 mutant A84F, with triolein as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 948
-
 mutant A84F, with 4-nitrophenyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 1057
-
 wild-type, with 4-nitrophenyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 1286
-
 wild-type, with tricaprylin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 2019
-
 mutant A84F, with tricaprylin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 3302
-
 wild-type, with tributyrin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola
3.1.1.74 4394
-
 mutant A84F, with tributyrin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic Alternaria brassicicola

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.74 4-nitrophenyl butyrate + H2O
-
 Alternaria brassicicola 4-nitrophenol + butyrate
-
 ?
3.1.1.74 4-nitrophenyl palmitate + H2O
-
 Alternaria brassicicola 4-nitrophenol + palmitate
-
 ?
3.1.1.74 ethyl butyrate + H2O
-
 Alternaria brassicicola butyric acid + ethanol
-
 ?
3.1.1.74 ethyl caprylate + H2O
-
 Alternaria brassicicola caprylic acid + ethanol
-
 ?
3.1.1.74 methyl acetate + H2O
-
 Alternaria brassicicola acetic acid + methanol
-
 ?
3.1.1.74 methyl butyrate + H2O
-
 Alternaria brassicicola butyric acid + methanol
-
 ?
3.1.1.74 methyl caproate + H2O
-
 Alternaria brassicicola caproic acid + methanol
-
 ?
3.1.1.74 methyl caprylate + H2O
-
 Alternaria brassicicola caprylic acid + methanol
-
 ?
3.1.1.74 methyl decanoate + H2O
-
 Alternaria brassicicola decanoic acid + methanol
-
 ?
3.1.1.74 methyl laurate + H2O
-
 Alternaria brassicicola lauric acid + methanol
-
 ?
3.1.1.74 methyl myristate + H2O
-
 Alternaria brassicicola myristic acid + methanol
-
 ?
3.1.1.74 methyl propionate + H2O
-
 Alternaria brassicicola propionic acid + methanol
-
 ?
3.1.1.74 tributyrin + H2O
-
 Alternaria brassicicola ?
-
 ?
3.1.1.74 tricaprylin + H2O
-
 Alternaria brassicicola ?
-
 ?
3.1.1.74 triolein + H2O
-
 Alternaria brassicicola ?
-
 ?
3.1.1.74 tripalmitin + H2O
-
 Alternaria brassicicola ?
-
 ?
3.1.1.74 tristearin + H2O
-
 Alternaria brassicicola ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.74 monomer 1 * 24000, SDS-PAGE, glycosylated CUTAB1. 1 * 20000, SDS-PAGE, non-glycosylated CUTAB1 Alternaria brassicicola

Synonyms

EC Number Synonyms Comment Organism
3.1.1.74 CUTAB1
-
 Alternaria brassicicola
3.1.1.74 cutinase
-
 Alternaria brassicicola

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.74 40
-
-
 Alternaria brassicicola

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.74 40
-
 retains 84% of its initial activity at 40°C and pH 8.0 for 5 days Alternaria brassicicola

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.74 7 9 at both, room temperature and 40°C Alternaria brassicicola