Information on EC 1.2.1.16 - succinate-semialdehyde dehydrogenase [NAD(P)+]

-

Co2+

-

K+

-

slight activation

Mg2+

-

100% activity at 10 mM Mg2+. With Mg2+ there is a 4fold decrease in the Km value for succinate semialdehyde and a 6fold decrease in the Km value for NADP+

Mn2+

-

Ni2+

-

additional information

3-tolualdehyde

Drosophila melanogaster

-

only the aldehyde forms and not the gem-diol forms of the inhibitor 3-tolualdehyde bind to the enzyme

690736

5,5'-dithiobis(2-nitrobenzoate)

-

5,5-dipropylpentanoate

-

coenzyme A

1 mM, 15% inhibition

dipropylacetate

-

glyoxylate

Malonic semialdehyde

N-ethylmaleimide

-

NADPH

p-chloromercuribenzoate

-

p-Hydroxybenzaldehyde

-

288048

potassium arsenite

potassium phosphate buffer

-

succinate semialdehyde

4 entries

Succinic semialdehyde

Zn2+

-

additional information

-

2-mercaptoethanol

4-aminobutyrate

induction during growth with 4-aminobutyrate (GABA)

4-aminobutyric acid

-

induction

cysteine

-

288046

dithiothreitol

5 entries

gamma-aminobutyrate

-

induces GabD dehydrogenase

692836

glutathione

-

288046

sulfhydryl reagents

0.1 - 0.3

2-oxoglutarate semialdehyde

0.041

3-sulfopropanal

Km apparent value

0.0069 - 0.035

benzaldehyde

6 entries

0.0092 - 17

NAD+

26 entries

0.01 - 27

NADP+

21 entries

0.002 - 20.5

succinate semialdehyde

36 entries

0.0011 - 0.03

Succinic semialdehyde

additional information

-

7.1 - 27.3

2-oxoglutarate semialdehyde

6.4 - 57

benzaldehyde

1.94 - 176

NAD+

20 entries

1.3 - 46

NADP+

17 entries

0.3 - 164

succinate semialdehyde

28 entries

16.8 - 22.6

Succinic semialdehyde

134.1 - 215.3

2-oxoglutarate semialdehyde

3.8 - 880

NAD+

16 entries

0.077 - 30660

NADP+

14 entries

0.36 - 14700

succinate semialdehyde

27 entries

1789 - 3454

Succinic semialdehyde

0.0628 - 470.6

succinate semialdehyde

0.0017

-

wild type, value below, NH4+ used as N-source or uga2 mutant unable to grow on 4-aminobutyric acid, glutamate and 4-aminobutyric acid used as N-source

0.0025

-

wild type, glutamate used as N-source

0.0033

-

uga3 mutant unable to grow on 4-aminobutyric acid, glutamate and 4-aminobutyric acid used as N-source

0.0038

H16-strain, coenzyme NAD+

0.0097

SK4580-strain, coenzyme NAD+

0.0105

HF39-strain, coenzyme NAD+

0.0172

SHF3-strain, coenzyme NAD+

0.021

-

wild type, succinate used as C-source, gamma-aminobutyrate and NH3 used as N-source

0.023

0.0235

SK4580-strain, coenzyme NADP+

0.0253

H16-strain, coenzyme NADP+

0.026

0.028

-

bis(3-aminopropyl)-amine used as C-source and N-source

0.029

-

spermidine used as C-source and N-source

0.03

-

bis(3-aminopropyl)-amine and glucose, glucose and NH3 or ribose and NH3 used as C-source and N-source

0.0319

SK4575-strain, coenzyme NAD+

0.032

-

gamma-aminobutyric acid and glucose used as C-source and N-source

0.033

-

spermidine and glucose used as C-source and N-source

0.034

-

wild type, 4-hydroxyphenylacetate used as C-source and NH3 used as N-source

0.035

-

gamma-aminobutyric acid used as C-source and N-source

0.0355

SK4575-strain, coenzyme NADP+

0.036

-

CS101A strain, unable to utilize 4-aminobutyrate as N-source

0.038

-

putrescine used as C-source and N-source

0.0409

HF39-strain, coenzyme NADP+

0.045

-

gamma-hydroxybutyric acid and NH3 used as C-source and N-source

0.0457

SHF3-strain, coenzyme NADP+

0.06

-

4-hydroxyphenylacetic acid and glucose used as carbon source, NH3 used as N-source

0.07

-

succinate used as carbon source, NH3 used as N-source

0.073

-

nutrient broth used as carbon source, NADP-dependent form

0.075

0.078

-

glycerol used as carbon source, 4-aminobutyric acid used as N-source

0.08

-

4-hydroxyphenylacetic acid used as carbon source, NH3 used as N-source

0.085

-

4-aminobutyric acid used as carbon source, NH3 used as N-source

0.089

-

mutant, glucose used as C-source and NH3 used as N-source

0.091

-

4-hydroxyphenylacetic acid, glucose and cAMP used as carbon source, NH3 used as N-source

0.097

-

mutant, succinate used as C-source and NH3 used as N-source

0.105

-

wild type, succinate used as C-source and gamma-aminobutyrate used as N-source

0.125

-

uga1 mutant unable to grow on 4-aminobutyric acid, glutamate and 4-aminobutyric acid used as N-source

0.13

-

4-hydroxyphenylacetic acid and cAMP used as carbon source, NH3 used as N-source

0.319

-

mutant, gamma-aminobutyrate used as C-source and NH3 used as N-source

0.351

-

mutant, succinate used as C-source and gamma-aminobutyrate used as N-source

0.38

-

wild type, glutamate and 4-aminobutyric acid used as N-source

0.395

-

mutant, glucose used as C-source and gamma-aminobutyrate used as N-source

0.408

-

mutant, gamma-aminobutyrate used as C-source and as N-source

0.5 - 0.66

-

wild type, 4-aminobutyric acid used as N-source

0.731

-

CS101G2 strain, unable to utilize 4-aminobutyrate as N-source

0.797

-

S-5 strain, unable to utilize 4-aminobutyrate as N-source

1.012

-

S-9 strain, unable to utilize 4-aminobutyrate as N-source

1.023

-

L-18 strain, unable to utilize 4-aminobutyrate as N-source

1.038

-

D-8 strain, unable to utilize 4-aminobutyrate as N-source

1.08

-

M-20 strain, unable to utilize 4-aminobutyrate as N-source

1.264

-

CS101B strain, able to utilize 4-aminobutyrate as N-source

7.94

-

8.5

40.7

-

pH 8.0, 30°C

additional information

10 entries

-

specific activity of 0.3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 0.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is succinale semialdehyde and NAD+; specific activity of 0.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 0.6 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 0.7 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 10 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 2.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 5.0 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is succinale semialdehyde and NAD+; specific activity of 5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 7.0 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 7.3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is succinale semialdehyde and NAD+

specific activity of 0.3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 0.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is succinale semialdehyde and NAD+; specific activity of 0.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 0.6 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with ammonium as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 0.7 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 10 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 2.5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is sulfopropanal and NADP+; specific activity of 3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 5.0 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with 4-aminobutyrate as the sole nitrogen source, substrate is succinale semialdehyde and NAD+; specific activity of 5 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is sulfopropanal and NAD+; specific activity of 7.0 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is succinale semialdehyde and NADP+; specific activity of 7.3 mkat/kg protein is detected in crude cell extracts of Cupriavidus necator grown with homotaurine as the sole nitrogen source, substrate is succinale semialdehyde and NAD+

7

7.5 - 7.8

-

Salmonella enterica subsp. enterica serovar Typhimurium

7.5 - 9

-

8.4

-

8.5

8.7

-

9 - 9.5

-

9.7

-

NAD+

9.9

-

NADP+

11

in 50 mM glycine-NaOH buffer

5 - 10

-

KM value for NAD+ increases from 0.3 to 1 mM as the pH changes from pH 5 to 10. The pH profile data are obtained at a NAD+ concentration of 5 mM, essentially saturating across the entire pH range. Similarly, data for the NADP+ pH profile are obtained at 10 mM NADP+

5.5 - 10

-

the enzyme cannot be assayed below pH 5.5, due to precipitation and loss of activity

7 - 9.3

-

at pH 7.0 and 9.3: about 30% of activity maximum

7.5 - 9.5

-

8 - 10.5

specific activity increases steadily to a broad optimum

8.7 - 10.6

-

with NAD+, at pH 8.7: 45% of activity maximum, at pH 10.6: about 50% of activity maximum

8.7 - 10.9

-

with NADP+, at pH 8.7: about 30% of activity maximum, at pH 10.9: about 65% of activity maximum

additional information

-

The pH profiles are bell-shaped indicating that two ionizable groups are involved in the catalytic mechanism.

30

31

-

35 - 45

-

90

-

-

3ek1, download, 3D-view

SCOPe (3ek1)

CATH (3ek1)

Brucella abortus (strain 2308)

Q2YRI3

4v6h, download, 3D-view

SCOPe (4v6h)

CATH (4v6h)

Burkholderia pseudomallei (strain 1710b)

Q3JS51

3jz4, download, 3D-view

SCOPe (3jz4)

CATH (3jz4)

Escherichia coli (strain K12)

P25526

3ros, download, 3D-view

SCOPe (3ros)

CATH (3ros)

Lactobacillus acidophilus (strain ATCC 700396 / NCK56 / N2 / NCFM)

Q5FIM9

3rh9, download, 3D-view

SCOPe (3rh9)

CATH (3rh9)

Marinobacter hydrocarbonoclasticus (strain ATCC 700491 / DSM 11845 / VT8)

A1U6U7

45000

-

2 * 45000, in solution, SDS-PAGE,

47400

-

calculated from amino acid sequence

49000

51600

52000

53000

-

2 * 53000, SDS-PAGE

59000

-

4 * 59000, SDS-PAGE

Salmonella enterica subsp. enterica serovar Typhimurium

84000

-

gel filtration

90000

-

Dimer is determined by gel filtration whereas the molecular mass of the enzyme is calculated by comparison to the relative mobility of the standard proteins.; SDS-PAGE

100000

-

gel filtration

114000

-

gel filtration

150000

-

gel filtration

180000

gel filtration

200000

-

gel filtration

275000

-

gel filtration

300000

-

gel filtration

?

Sulfolobus solfataricus

-

x * 51600, SDS-PAGE

dimer

homodimer

tetramer

additional information

Salmonella enterica subsp. enterica serovar Typhimurium

apo-form and in complex with NAD+, hanging drop vapor diffusion method, using 2 M sodium formate and 0.1 M sodium acetate (pH 4.6), at 22°C

-

Salmonella enterica subsp. enterica serovar Typhimurium

structures of apoenzyme and in complex with NAD+, to 1.85 and 1.9 A resolution. Two domain protein with the active site located in the interdomain interface. The NAD+ molecule is bound in the long channel with its nicotinamide ring positioned close to the side chain of the catalytic Cys268

-

741378

apoenzyme and in complex with NAD+, hanging drop vapor diffusion method, using 0.1 M MES monohydrate pH 5.8-6.1 and 18-23% (w/v) polyethylene glycol 4000

-

723864

purified recombinant SpSSADH in apo-form and in a binary complex with NADP+, hanging drop vapor diffusion method, for the apoform crystals: mixing 0.001 ml of protein solution with 0.001 ml of reservoir solution containing 0.1 M MES monohydrate, pH 5.8-6.1, and 18-23% w/v PEG 4000, and equilibration over 0.5 ml reservoir solution, for the binary complex crystals: mixing of protein solution with NADP+ in a 1:10 molar ratio with reservoir solution containing 0.1 M sodium acetate trihydrate, pH 4.6, and 2 M ammonium sulfate, 22°C, X-ray diffraction structure determination and analysis at 1.6 A and 2.1 A resolutions, respectively, molecular replacement method using the structure of Escherichia coli SSADH, PDB ID 3JZ4, as search model

A0A0J9X1MB

743290

4.5

enzyme is almost inactive

698627

5 - 10

-

the Km value for NAD+ increases from 0.3 to 1 mM as the pH changes from pH 5 to 10

685387

6.5 - 7.1

-

highest stability

288041, 288046

0 - 2

-

substancial progressive loss of activity, sucrose fractions

288048

25

-

substancial progressive loss of activity, sucrose fractions

288048

50

60

-

up to, in presence of 30% glycerol

dithiothreitol is necessary both to maintain the activity of BADH and to prevent oligimerization of the enzyme

-

enzyme is labile (50% loss of activity overnight) but can be largely stabilized (in crude extract only) by the addition of 10 mM dithiothreitol 1.2.1.16

ethyleneglycol, 30%, stabilizes

-

glycerol, 30%, stabilizes

-

there is no significant loss of catalytic activity, after up to 40 min of incubation of isoform GabD1 with 1 mM H2O2 at pH 7.4

-

724115

2°C, pH 7.0, 10% loss of activity per week

-

288041, 288046

3°C or -15°C, overnight, 100% loss of activity, without mercaptoethanol

-

287170

5°C, complete inactivation after 2 days

-

Addition of 2 mM DTT to both stacking and resolving gels results in a decrease in the protein laddering effect, and both BADH and BADH-His are present as near-homogeneous species.

-

-

-

288041, 288046

ammonium sulfate fractionation and DEAE-cellulose

-

ammonium sulfate fractionation, phenyl-Sepharose column chromatography, and Q-Sepharose column chromatography

Pseudomonas stutzeri

-

678422

by ammonium sulfate precipitation and anion exchange chromatography and gel filtration

chromatography on Celite 535

-

288050

chromatography on Sephacryl S-300, DEAE-Sephacel and Sepharose 4B

-

column chromatography on DEAE-cellulose, Sephadex G-200 and DEAE-Sephadex

-

DEAE-cellulose chromatography

-

287170

His-tagged GabD dehydrogenase purified from cell extracts with HiTrap affinity columns

-

692836

Ni-NTA resin column chromatography

-

Salmonella enterica subsp. enterica serovar Typhimurium

Ni-NTA resin column chromatography and Superdex 75 gel filtration

-

Ni2+-agarose chromatography; Purification of his-tagged proteins by using a standard Ni2+-agarose chromatography protocol.

-

polyanionic resin Polyanion S1-HR5/5

-

recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography and gel filtration

Bacillus subtilis

-

743077

recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration

A0A0J9X1MB

743290

UNO Q-12 column chromatography

-

UNO Q-12 column chromatography and Superdex 200 gel filtration

Sulfolobus solfataricus

-

-

Drosophila melanogaster

-

690736

expressed in Escherichia coli BL21 (DE3) cells

-

723864

expressed in Escherichia coli BL21 (DE3)-RIL cells

Sulfolobus solfataricus

-

Salmonella enterica subsp. enterica serovar Typhimurium

expressed in Escherichia coli BL21 cells

-

expressed in Escherichia coli BL21(DE3) cells

Pseudomonas stutzeri

-

678422

expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells

-

expressed in Escherichia coli strain BL21(DE3)pLysS; Protein expression in Escherichia coli strain JM109. Single colonies are picked and the DNA is isolated and screened for the desired mutation either by restriction analysis (C103A and C249A) or by sequencing (C140A and C220A). The overall fidelity of the PCR amplification and the presence of the desired mutations are confirmed by sequencing. The plasmids containing the mutated genes are then transformed into Escherichia coli strain BL21(DE3)pLysS for protein expression.

-

expression in Escherichia coli

gabD gene cloned, expression in Escherichia coli

gene ssadh, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)

A0A0J9X1MB

743290

overexpressed as His-tagged GabD

-

692836

recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS

Bacillus subtilis

-

743077

expression is induced by exogenous succinic semialdehyde

-

induction by growth on 4-hydroxyphenylacetate

-

transcript is detected inducibly in both homotaurine and GABA-grown cells

C311A

Drosophila melanogaster

-

mutant is still soluble but unable to catalyze succinate semialdehyde oxidation. Mutation leads to an inactive product binding both succinate semialdehyde aldehyde and gem-diol

690736

C102A

-

mutant exhibits BADH activity between 50 and 75% that of the wild type enzyme

C103A

-

enzyme is inactive