NAD-dependent glutamate dehydrogenase from Pyrobaculum islandicum

Enzyme Description

Species

Pyrobaculum islandicum ↗

Extremophile

Yes "hyperthermophilic" organism

EC Number

1.4.1.2 ↗

Sequence

Length: 421 amino acids

MERTGFLEYVLNYVKKGVELGGFPEDFYKILSRPRRVLIVNIPVRLDGGGFEVFEGYRVQHCDVLGPYKGGVRFHPEVTLADDVALAILMTLKNSLAGLPYGGAKGAVRVDPKKLSQRELEELSRGYARAIAPLIGDVVDIPAPDVGTNAQIMAWMVDEYSKIKGYNVPGVFTSKPPELWGNPVREYATGFGVAVATREMAKKLWGGIEGKTVAIQGMGNVGRWTAYWLEKMGAKVIAVSDINGVAYRKEGLNVELIQKNKGLTGPALVELFTTKDNAEFVKNPDAIFKLDVDIFVPAAIENVIRGDNAGLVKARLVVEGANGPTTPEAERILYERGVVVVPDILANAGGVIMSYLEWVENLQWYIWDEEETRKRLENIMVNNVERVYKRWQREKGWTMRDAAIVTALERIYNAMKIRGWI

Chizu Kujo et al. (1998) — Enzymological Characteristics of the Hyperthermostable NAD-Dependent Glutamate Dehydrogenase from the Archaeon Pyrobaculum islandicum and Effects of Denaturants and Organic Solvents

Applied and Environmental Microbiology · doi:10.1128/aem.64.6.2152-2157.1998 ↗ · Activity - Classical Stability - Incubation

67 measurements ▼

Database ID

UniProt: Q9Y8I4 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Purified, archaeon Pyrobaculum islandicum

Experimental Data (67 measurements)

67 measurements — page 3 of 4

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	20%	100	104	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	30%	100	121	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	40%	100	107	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	50%	100	21	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100	100	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	100	111	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	100	111	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	100	109	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	50%	100	104	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	40%	100	35	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	30%	100	45	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	20%	100	96	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	10%	100	108	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	50%	100	22	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	40%	100	106	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	30%	100	106	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	20%	100	119	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+ , α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase

Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	40%	100	110	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	5%	100	217	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	10%	100	228	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	15%	100	162	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100	38	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	30%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	10%	100	140	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	20%	100	158	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100	145	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	20%	100	196	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	50%	100	49	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethylformamide (DMF)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100	76	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100	61	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100	46	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100	22	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100	19	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	25%	100	46	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	30%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	10%	100	185	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	30%	100	190	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	40%	100	189	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	50%	100	159	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	60%	100	49	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	10%	100	76	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	20%	100	60	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	40%	100	22	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	50%	100	19	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	5%	100	172	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	10%	100	221	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	15%	100	240	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	20%	100	194	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	30%	100	46	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Classical aqueous control (in %)
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	50%	100	30	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Tetrahydrofuran (THF)	10%	100	108	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Tetrahydrofuran (THF)	20%	100	83	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Tetrahydrofuran (THF)	30%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	10%	100	107	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	20%	100	104	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	30%	100	121	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	40%	100	107	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	50%	100	21	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100	100	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	20%	100	111	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	30%	100	111	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	40%	100	109	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	50%	100	104	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	40%	100	35	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	30%	100	45	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	20%	100	96	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	10%	100	108	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	50%	100	22	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	40%	100	106	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	30%	100	106	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	20%	100	119	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Methanol	10%	100	100	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	60%	100	0	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	50%	100	24	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	40%	100	101	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	30%	100	124	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	20%	100	122	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (10 minutes at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase	Ethanol	10%	100	107	%	0.2 micoM glycine-KOH buffer	9.7	50°C	10 µmol L-glutamic acid	NH4+, α-ketoglutarate	1.25 µM NAD	—	Non-incubated control (in %), assay in aqueous phase

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Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability — Incubation