Transglutaminase from Streptomyces mobaraensis

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 339 amino acids
DSDDRVTPPAEPLDRMPDPYRPSYGRAETVVNNYIRKWQQVYSHRDGRKQQMTEEQREWLSYGCVGVTWVNSGQYPTNRLAFASFDEDRFKNELKNGRPRSGETRAEFEGRVAKESFDEEKGFQRAREVASVMNRALENAHDESAYLDNLKKELANGNDALRNEDARSPFYSALRNTPSFKERNGGNHDPSRMKAVIYSKHFWSGQDRSSSADKRKYGDPDAFRPAPGTGLVDMSRDRNIPRSPTSPGEGFVNFDYGWFGAQTEADADKTVWTHGNHYHAPNGSLGAMHVYESKFRNWSEGYSDFDRGAYVITFIPKSWNTAPDKVKQGWPLEHHHHHH
Gabe Javitt et al. (2017) β€” Constitutive expression of active microbial transglutaminase in Escherichia coli and comparative characterization to a known variant
BMC Biotechnology  Β· doi:10.1186/s12896-017-0339-4 β†—  Β· Activity - Classical Activity + Stability - Incubation
104 measurements
Database ID
Sequence Annotation
Explicit - Provided GenBank Accession Number (first residue absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (104 measurements)

104 measurements β€” page 4 of 6
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 20% 100 88 69 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 30% 100 67 72 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 40% 100 38 64 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 50% 100 39 40 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 60% 100 54 57 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Isopropanol 70% 100 65 72 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 10% 100 99 94 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 20% 100 94 94 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 30% 100 116 99 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 40% 100 79 82 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 50% 100 54 72 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 60% 100 20 21 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Methanol 70% 100 23 13 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 11% 100 58 64 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 21% 100 77 62 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 31% 100 69 57 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 41% 100 56 40 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 51% 100 24 35 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 61% 100 21 42 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
S2P Activity - Classical Activity measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 70% 100 4 0 % 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA 6 37Β°C 53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine Glycine , Ξ³-glutamyl hydroxamate β€” β€” Classical aqueous control (in %), assay in the presence of organic solvent
1 … 3 4 5 6

Mutations in this dataset (2)

WT S2P

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 : Activity + 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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