Transglutaminase from Streptomyces mobaraensis

Enzyme Description

Species

Extremophile

EC Number

2.3.2.13 ↗

Sequence

Length: 339 amino acids

DSDDRVTPPAEPLDRMPDPYRPSYGRAETVVNNYIRKWQQVYSHRDGRKQQMTEEQREWLSYGCVGVTWVNSGQYPTNRLAFASFDEDRFKNELKNGRPRSGETRAEFEGRVAKESFDEEKGFQRAREVASVMNRALENAHDESAYLDNLKKELANGNDALRNEDARSPFYSALRNTPSFKERNGGNHDPSRMKAVIYSKHFWSGQDRSSSADKRKYGDPDAFRPAPGTGLVDMSRDRNIPRSPTSPGEGFVNFDYGWFGAQTEADADKTVWTHGNHYHAPNGSLGAMHVYESKFRNWSEGYSDFDRGAYVITFIPKSWNTAPDKVKQGWPLEHHHHHH

Gabe Javitt et al. (2017) — Constitutive expression of active microbial transglutaminase in Escherichia coli and comparative characterization to a known variant

BMC Biotechnology · doi:10.1186/s12896-017-0339-4 ↗ · Activity - Classical Activity + Stability - Incubation

104 measurements ▼

Database ID

UniParc: UPI0009C2BAFA ↗

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 — page 2 of 6

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	10%	100	—	58	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	20%	100	—	77	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	30%	100	—	69	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	40%	100	—	56	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	50%	100	—	24	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	60%	100	—	21	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	10%	100	—	108	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	20%	100	—	111	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	30%	100	—	116	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	40%	100	—	76	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Isopropanol	10%	100	—	65	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	59	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	42	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	23	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	111	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	128	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine , γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent

WT	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	Ethanol	10%	100	—	90	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	20%	100	—	59	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	30%	100	—	49	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	40%	100	—	34	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	50%	100	—	20	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	60%	100	—	24	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	Ethanol	70%	100	—	18	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	10%	100	—	88	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	10%	100	—	58	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	20%	100	—	77	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	30%	100	—	69	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	40%	100	—	56	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	50%	100	—	24	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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)	60%	100	—	21	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	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	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	10%	100	—	108	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	20%	100	—	111	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	30%	100	—	116	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	40%	100	—	76	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Isopropanol	10%	100	—	65	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	59	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	42	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	23	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	111	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	128	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	135	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	116	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	141	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	10%	100	—	122	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	20%	100	—	146	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	30%	100	—	155	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	40%	100	—	159	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	50%	100	—	143	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	60%	100	—	108	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	—	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated 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	Ethanol	10%	100	90	90	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	20%	100	59	61	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	30%	100	49	45	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	40%	100	34	33	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	50%	100	20	28	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	60%	100	24	16	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	Ethanol	70%	100	18	15	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	10%	100	88	78	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	20%	100	88	69	%	356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	6	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	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	37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Classical aqueous control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	10%	100	108	105	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	20%	100	111	99	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	30%	100	116	96	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Ethanol	40%	100	76	54	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), measured by absorbance spectrophotometry (colorimetric assay, FeCl3 reagant and hydroxamate reaction product absorbance measurement, 525 nm) in the presence of organic solvent	Isopropanol	10%	100	65	62	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	59	56	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	42	37	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	23	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	0	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	0	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	111	120	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	128	122	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	135	121	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	116	103	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	141	83	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	0	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	0	0	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	10%	100	122	117	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	20%	100	146	137	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	30%	100	155	143	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	40%	100	159	146	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	50%	100	143	124	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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)	60%	100	108	95	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent
S2P	Activity + Stability - Incubation	Activity after incubation (24 hours at 4°C), 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	0	4	%	Incubation: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA, Assay: 356 mM Tris-acetate buffer, 10 mM sodium phosphate buffer, 1.8 mM EDTA	Incubation: 6, Assay: 6	Incubation: 4°C, Assay: 37°C	53 mM benzyloxycarbonyl-glutaminyl-glycine, 352 mM Hydroxylamine	Glycine, γ-glutamyl hydroxamate	—	—	Water-incubated control (in %), assay in the presence of organic solvent

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Mutations in this dataset (2)

WT S2P

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 : Activity + 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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Transglutaminase from Streptomyces mobaraensis

Enzyme Description

Sequence

Experimental Data (104 measurements)

Visualization : Activity — Classical

Mutation Effect

Visualization : Activity + Stability — Incubation

Mutation Effect

Structure

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