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 2 of 4

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Extraction method	Data type	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Activity - Classical	Activity assayd by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	10 % v/v	100	185	%	Digital	Continuous	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 % v/v	100	190	%	Digital	Continuous	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 % v/v	100	189	%	Digital	Continuous	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 % v/v	100	159	%	Digital	Continuous	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 % v/v	100	49	%	Digital	Continuous	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 % v/v	100	76	%	Digital	Continuous	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 % v/v	100	60	%	Digital	Continuous	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 % v/v	100	22	%	Digital	Continuous	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 % v/v	100	19	%	Digital	Continuous	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 % v/v	100	0	%	Digital	Continuous	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 % v/v	100	172	%	Digital	Continuous	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 % v/v	100	221	%	Digital	Continuous	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 % v/v	100	240	%	Digital	Continuous	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 % v/v	100	194	%	Digital	Continuous	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 % v/v	100	46	%	Digital	Continuous	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 % v/v	100	30	%	Digital	Continuous	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 % v/v	100	108	%	Digital	Continuous	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 % v/v	100	83	%	Digital	Continuous	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 % v/v	100	0	%	Digital	Continuous	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 % v/v	100	107	%	Digital	Continuous	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 % v/v

100

110

Digital

Continuous

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 % v/v

100

217

Digital

Continuous

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 % v/v

100

228

Digital

Continuous

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 % v/v

100

162

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

140

Digital

Continuous

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 % v/v

100

158

Digital

Continuous

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 % v/v

100

145

Digital

Continuous

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 % v/v

100

196

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

185

Digital

Continuous

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 % v/v

100

190

Digital

Continuous

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 % v/v

100

189

Digital

Continuous

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 % v/v

100

159

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

172

Digital

Continuous

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 % v/v

100

221

Digital

Continuous

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 % v/v

100

240

Digital

Continuous

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 % v/v

100

194

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

108

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

107

Digital

Continuous

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 % v/v

100

104

Digital

Continuous

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 % v/v

100

121

Digital

Continuous

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 % v/v

100

107

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

111

Digital

Continuous

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 % v/v

100

111

Digital

Continuous

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 % v/v

100

109

Digital

Continuous

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 % v/v

100

104

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

108

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

106

Digital

Continuous

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 % v/v

100

106

Digital

Continuous

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 % v/v

100

119

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

Digital

Continuous

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 % v/v

100

101

Digital

Continuous

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 % v/v

100

124

Digital

Continuous

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 % v/v

100

122

Digital

Continuous

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 % v/v

100

107

Digital

Continuous

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

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

Structure

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NAD-dependent glutamate dehydrogenase from Pyrobaculum islandicum

Enzyme Description

Sequence

Experimental Data (67 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Structure

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