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

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+

724115

Mn2+

Mycobacterium tuberculosis

-

724115

Ni2+

Mycobacterium tuberculosis

-

724115

additional information

2 entries

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)

Euglena gracilis

-

288042

5,5-dipropylpentanoate

-

coenzyme A

1 mM, 15% inhibition

dipropylacetate

-

glyoxylate

Malonic semialdehyde

N-ethylmaleimide

-

NADPH

p-chloromercuribenzoate

Euglena gracilis

-

288042

p-Hydroxybenzaldehyde

Rattus norvegicus

-

288048

potassium arsenite

3 entries

potassium phosphate buffer

Euglena gracilis

-

288042

succinate semialdehyde

3 entries

Succinic semialdehyde

2 entries

Zn2+

Euglena gracilis

-

288042

additional information

2 entries

-

2-mercaptoethanol

3 entries

4-aminobutyrate

Cupriavidus necator

Q0K2K1

induction during growth with 4-aminobutyrate (GABA)

698627

4-aminobutyric acid

Escherichia coli

-

induction

288039

cysteine

Pseudomonas fluorescens

-

288046

dithiothreitol

5 entries

gamma-aminobutyrate

Escherichia coli

-

induces GabD dehydrogenase

692836

glutathione

Pseudomonas fluorescens

-

288046

sulfhydryl reagents

2 entries

0.1 - 0.3

2-oxoglutarate semialdehyde

3 entries

0.041

3-sulfopropanal

Cupriavidus necator

Q0K2K1

Km apparent value

698627

0.0069 - 0.035

benzaldehyde

6 entries

0.0092 - 17

NAD+

24 entries

0.01 - 27

NADP+

19 entries

0.002 - 20.5

succinate semialdehyde

34 entries

0.0011 - 0.03

Succinic semialdehyde

3 entries

7.1 - 27.3

2-oxoglutarate semialdehyde

3 entries

6.4 - 57

benzaldehyde

2 entries

2.7 - 176

NAD+

18 entries

1.3 - 46

NADP+

15 entries

0.3 - 164

succinate semialdehyde

26 entries

16.8 - 22.6

Succinic semialdehyde

3 entries

134.1 - 215.3

2-oxoglutarate semialdehyde

3 entries

3.8 - 880

NAD+

14 entries

0.077 - 30660

NADP+

12 entries

0.36 - 14700

succinate semialdehyde

25 entries

1789 - 3454

Succinic semialdehyde

3 entries

0.0628

succinate semialdehyde

Mycobacterium tuberculosis

-

in the absence of Mg2+, at pH 7.5 and 37°C

724115

0.0017

Saccharomyces cerevisiae

-

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

288037

0.0025

Saccharomyces cerevisiae

-

wild type, glutamate used as N-source

288037

0.0033

Saccharomyces cerevisiae

-

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

288037

0.0038

Cupriavidus necator

Q9RBF6

H16-strain, coenzyme NAD+

288054

0.0097

Cupriavidus necator

Q9RBF6

SK4580-strain, coenzyme NAD+

288054

0.0105

Cupriavidus necator

Q9RBF6

HF39-strain, coenzyme NAD+

288054

0.0172

Cupriavidus necator

Q9RBF6

SHF3-strain, coenzyme NAD+

288054

0.021

Escherichia coli

-

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

0.023

0.0235

SK4580-strain, coenzyme NADP+

288054

0.0253

Cupriavidus necator

Q9RBF6

H16-strain, coenzyme NADP+

288054

0.026

2 entries

0.028

Pseudomonas sp.

-

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

288045

0.029

Pseudomonas sp.

-

spermidine used as C-source and N-source

288045

0.03

Pseudomonas sp.

-

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

288045

0.0319

Cupriavidus necator

Q9RBF6

SK4575-strain, coenzyme NAD+

288054

0.032

Pseudomonas sp.

-

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

288045

0.033

Pseudomonas sp.

-

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

288045

0.034

Escherichia coli

-

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

288039

0.035

Pseudomonas sp.

-

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

288045

0.0355

Cupriavidus necator

Q9RBF6

SK4575-strain, coenzyme NADP+

288054

0.036

Escherichia coli

-

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

288051

0.038

Pseudomonas sp.

-

putrescine used as C-source and N-source

288045

0.0409

Cupriavidus necator

Q9RBF6

HF39-strain, coenzyme NADP+

288054

0.045

Pseudomonas sp.

-

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

288045

0.0457

Cupriavidus necator

Q9RBF6

SHF3-strain, coenzyme NADP+

288054

0.06

Klebsiella pneumoniae

-

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

288038

0.07

Klebsiella pneumoniae

-

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

288038

0.073

Klebsiella pneumoniae

-

nutrient broth used as carbon source, NADP-dependent form

288038

0.075

2 entries

0.078

Klebsiella pneumoniae

-

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

288038

0.08

Klebsiella pneumoniae

-

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

288038

0.085

Klebsiella pneumoniae

-

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

288038

0.089

Escherichia coli

-

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

288039

0.091

Klebsiella pneumoniae

-

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

288038

0.097

Escherichia coli

-

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

288039

0.105

Escherichia coli

-

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

288039

0.125

Saccharomyces cerevisiae

-

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

288037

0.13

Klebsiella pneumoniae

-

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

288038

0.319

Escherichia coli

-

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

288039

0.351

Escherichia coli

-

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

288039

0.38

Saccharomyces cerevisiae

-

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

288037

0.395

Escherichia coli

-

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

288039

0.408

Escherichia coli

-

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

288039

0.5 - 0.66

Saccharomyces cerevisiae

-

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

288037

0.731

Escherichia coli

-

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

288051

0.797

Escherichia coli

-

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

288051

1.012

Escherichia coli

-

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

288051

1.023

Escherichia coli

-

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

288051

1.038

Escherichia coli

-

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

288051

1.08

Escherichia coli

-

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

288051

1.264

Escherichia coli

-

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

7.94

-

8.5

40.7

-

pH 8.0, 30°C

707583

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+

698627

Q0K2K1

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

Klebsiella pneumoniae

-

288038

7.5 - 9

Salmonella enterica subsp. enterica serovar Typhimurium

-

726463

8.4

Saccharomyces cerevisiae

-

8.5

8.7

-

9 - 9.5

-

9.7

Rhodobacter sphaeroides

-

NAD+

288044

9.9

Rhodobacter sphaeroides

-

NADP+

288044

11

Cupriavidus necator

Q9RBF6

in 50 mM glycine-NaOH buffer

288054

5 - 10

Pseudomonas putida

-

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+

685387

5.5 - 10

Mycobacterium tuberculosis

-

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

724115

7 - 9.3

Pseudomonas fluorescens

-

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

698627

8.7 - 10.6

Rhodobacter sphaeroides

-

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

288044

8.7 - 10.9

Rhodobacter sphaeroides

-

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

288044

additional information

Pseudomonas putida

-

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

685387

30

Pseudomonas putida

-

assay at

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

Pseudomonas putida

-

2 * 45000, in solution, SDS-PAGE,

685387

47400

Pseudomonas putida

-

calculated from amino acid sequence

685387

49000

2 entries

51600

Escherichia coli

-

2 * 51600, SDS-PAGE

722211

52000

2 entries

53000

Pseudomonas putida

-

2 * 53000, SDS-PAGE

707583

59000

Klebsiella pneumoniae

-

4 * 59000, SDS-PAGE

288038

84000

Salmonella enterica subsp. enterica serovar Typhimurium

-

gel filtration

726463

90000

Pseudomonas putida

-

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

685387

100000

Pseudomonas putida

-

gel filtration

707583

114000

Escherichia coli

-

gel filtration

722211

150000

Euglena gracilis

-

gel filtration

180000

gel filtration

200000

-

gel filtration

288039

275000

Klebsiella pneumoniae

-

gel filtration

288038

300000

Klebsiella pneumoniae

-

gel filtration

288038

?

Sulfolobus solfataricus

-

x * 51600, SDS-PAGE

dimer

homodimer

tetramer

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

-

726463

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

Salmonella enterica subsp. enterica serovar Typhimurium

-

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

Streptococcus pyogenes

-

723864

4.5

Cupriavidus necator

Q0K2K1

enzyme is almost inactive

698627

5 - 10

Pseudomonas putida

-

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

Pseudomonas fluorescens

-

highest stability

288041, 288046

0 - 2

Rattus norvegicus

-

substancial progressive loss of activity, sucrose fractions

288048

25

Rattus norvegicus

-

substancial progressive loss of activity, sucrose fractions

288048

50

2 entries

60

Euglena gracilis

-

up to, in presence of 30% glycerol

288042

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

Pseudomonas putida

-

685387

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

Cupriavidus necator

Q0K2K1

698627

ethyleneglycol, 30%, stabilizes

Euglena gracilis

-

288042

glycerol, 30%, stabilizes

Euglena gracilis

-

288042

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

Mycobacterium tuberculosis

-

724115

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

Pseudomonas fluorescens

-

288041, 288046

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

Pseudomonas sp.

-

287170

5°C, complete inactivation after 2 days

Euglena gracilis

-

288042

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.

Pseudomonas putida

-

685387

-

Pseudomonas fluorescens

-

288041, 288046

ammonium sulfate fractionation and DEAE-cellulose

Euglena gracilis

-

288042

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

Cupriavidus necator

Q0K2K1

698627

chromatography on Celite 535

Rattus norvegicus

-

288050

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

Klebsiella pneumoniae

-

288038

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

Rhodobacter sphaeroides

-

288044

DEAE-cellulose chromatography

Pseudomonas sp.

-

287170

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

Escherichia coli

-

692836

Ni-NTA resin column chromatography

Mycobacterium tuberculosis

-

724115

Ni-NTA resin column chromatography and Superdex 75 gel filtration

Salmonella enterica subsp. enterica serovar Typhimurium

-

726463

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

Pseudomonas putida

-

685387

polyanionic resin Polyanion S1-HR5/5

Saccharomyces cerevisiae

-

288037

UNO Q-12 column chromatography

Escherichia coli

-

722211

UNO Q-12 column chromatography and Superdex 200 gel filtration

Sulfolobus solfataricus

-

722211

-

Drosophila melanogaster

-

690736

expressed in Escherichia coli BL21 (DE3) cells

Streptococcus pyogenes

-

723864

expressed in Escherichia coli BL21 (DE3)-RIL cells

Sulfolobus solfataricus

-

722211

expressed in Escherichia coli BL21 cells

Salmonella enterica subsp. enterica serovar Typhimurium

-

726463

expressed in Escherichia coli BL21(DE3) cells

Pseudomonas stutzeri

-

678422

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

Escherichia coli

-

722211

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.

Pseudomonas putida

-

685387

expression in Escherichia coli

2 entries

gabD gene cloned, expression in Escherichia coli

Cupriavidus necator

Q9RBF6

288054

overexpressed as His-tagged GabD

Escherichia coli

-

692836

expression is induced by exogenous succinic semialdehyde

Klebsiella pneumoniae

-

288038

induction by growth on 4-hydroxyphenylacetate

Pseudomonas putida

-

707583

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

Cupriavidus necator

Q0K2K1

698627

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

Pseudomonas putida

-

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

685387

C103A

Pseudomonas putida

-

enzyme is inactive

685387

C140A