Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Bifidobacterium catenulatum subsp. kashiwanohense |
gene sarD, genetic structure | Bilophila wadsworthia |
gene tauF, genetic structure, sequence comparisons, recombinant expression as His6-MBP-tagged enzyme in Escherichia coli strain BL21(DE3) from HMT-GGG-BkTauF plasmid | Bifidobacterium catenulatum subsp. kashiwanohense |
Crystallization (Comment) | Organism |
---|---|
purifed recombinant BkTauF in the apo form and in a binary complex with NAD+, hanging drop vapor diffusion method, mixing of 10 mg/ml protein in 20 mM Tris, pH 7.5, 200 mM KCl, 1 mM DTT, with reservoir solution containing 0.2 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, and 25% w/v PEG 3350, 22°C, method optimization, X-ray diffraction structure determination and analysis at 1.9 and 3.0 A resolution, respectively, molecular replacement using structure with PDB ID 1VHD as a search model, modeling | Bifidobacterium catenulatum subsp. kashiwanohense |
structures in the apo form and in a binary complex with NAD+ at 1.9 and 3.0 A resolution, respectively | Bifidobacterium catenulatum subsp. kashiwanohense |
Protein Variants | Comment | Organism |
---|---|---|
F252A | site-directed mutagenesis, the mutant shows 78% reduced activity for isethionate oxidation compared to wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense |
F252A | mutant retains approximately 28% of the wild-type activity | Bifidobacterium catenulatum subsp. kashiwanohense |
F265A | almost complete loss of activity | Bifidobacterium catenulatum subsp. kashiwanohense |
F265A | site-directed mutagenesis, the mutant shows highly reduced activity for isethionate oxidation compared to wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense |
T257A | almost complete loss of activity | Bifidobacterium catenulatum subsp. kashiwanohense |
T257A | site-directed mutagenesis, the mutant shows highly reduced activity for isethionate oxidation compared to wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | complete inhibition; complete loss of activity. Activity is recovered up to 85% by addition of Zn2+ and 15% by addition of Fe2+ | Bifidobacterium catenulatum subsp. kashiwanohense |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.06 | - |
NADH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.06 | - |
NADH | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.08 | - |
NAD+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.08 | - |
NAD+ | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.5 | - |
2-sulfoacetaldehyde | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.5 | - |
2-sulfoacetaldehyde | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.73 | - |
NADPH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.77 | - |
NADP+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
4.44 | - |
isethionate | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
4.44 | - |
isethionate | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | activates, can partially substitute for Zn2+ | Bifidobacterium catenulatum subsp. kashiwanohense | |
additional information | catalytic activity of BkTauF is abolished by chelation with EDTA, and recovered up to 85% by addition of Zn2+ and 15% by addition of Fe2+, suggesting that the physiological metal cofactor for BkTauF is Zn2+ | Bifidobacterium catenulatum subsp. kashiwanohense | |
Zn2+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | |
Zn2+ | required | Bifidobacterium catenulatum subsp. kashiwanohense |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
83000 | - |
gel filtration | Bifidobacterium catenulatum subsp. kashiwanohense |
83000 | - |
recombinant enzyme, gel filtration | Bifidobacterium catenulatum subsp. kashiwanohense |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-sulfoacetaldehyde + NADH + H+ | Bilophila wadsworthia | - |
isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | Bifidobacterium catenulatum subsp. kashiwanohense | - |
isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | Bilophila wadsworthia 3_1_6 | - |
isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | - |
isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADPH + H+ | Bifidobacterium catenulatum subsp. kashiwanohense | - |
isethionate + NADP+ | - |
r | |
2-sulfoacetaldehyde + NADPH + H+ | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | - |
isethionate + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bifidobacterium catenulatum subsp. kashiwanohense | - |
- |
- |
Bifidobacterium catenulatum subsp. kashiwanohense | A0A0A7I0A5 | - |
- |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | - |
- |
- |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | A0A0A7I0A5 | - |
- |
Bilophila wadsworthia | E5Y946 | - |
- |
Bilophila wadsworthia 3_1_6 | E5Y946 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-MBP-tagged enzyme from Escherichia coli strain BL21(DE3) by streptomycin sulfate, dialysis, amylose affinity chromatography, followed by MBP-tag cleavage through TEV protease, dialysis, and nickel affinity chromatography, anion exchange chromatography, and gel filtration | Bifidobacterium catenulatum subsp. kashiwanohense |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-sulfoacetaldehyde + NADH + H+ | - |
Bilophila wadsworthia | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | the reduction reaction direction is preferred by the enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | - |
Bilophila wadsworthia 3_1_6 | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADH + H+ | the reduction reaction direction is preferred by the enzyme | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | isethionate + NAD+ | - |
r | |
2-sulfoacetaldehyde + NADPH + H+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | isethionate + NADP+ | - |
r | |
2-sulfoacetaldehyde + NADPH + H+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | isethionate + NADP+ | - |
r | |
3-hydroxypropane-1-sulfonate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | ? + NADH + H+ | - |
? | |
3-hydroxypropane-1-sulfonate + NAD+ | low activity | Bifidobacterium catenulatum subsp. kashiwanohense | ? + NADH + H+ | - |
? | |
3-hydroxypropane-1-sulfonate + NAD+ | low activity | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | ? + NADH + H+ | - |
? | |
3-hydroxypropane-1-sulfonate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | ? + NADH + H+ | - |
? | |
3-hydroxypropionate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | ? + NADH + H+ | - |
? | |
3-hydroxypropionate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | ? + NADH + H+ | - |
? | |
3-hydroxypropionic acid + NAD+ | moderate to low activity | Bifidobacterium catenulatum subsp. kashiwanohense | ? + NADH + H+ | - |
? | |
isethionate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | 2-sulfoacetaldehyde + NADH + H+ | - |
r | |
isethionate + NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | 2-sulfoacetaldehyde + NADH + H+ | - |
r | |
isethionate + NADP+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | 2-sulfoacetaldehyde + NADPH + H+ | - |
r | |
additional information | substrate specificity, no activity with ethanol, ethylene glycol, and ethanolamine | Bifidobacterium catenulatum subsp. kashiwanohense | ? | - |
- |
|
additional information | no substrates: ethanol, ethylene glycol, or ethanolamine | Bifidobacterium catenulatum subsp. kashiwanohense | ? | - |
- |
|
additional information | substrate specificity, no activity with ethanol, ethylene glycol, and ethanolamine | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | ? | - |
- |
|
additional information | no substrates: ethanol, ethylene glycol, or ethanolamine | Bifidobacterium catenulatum subsp. kashiwanohense PV20-2 | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 41000, SDS-PAGE | Bifidobacterium catenulatum subsp. kashiwanohense |
homodimer | 2 * 41000, recombinant enzyme, SDS-PAGE | Bifidobacterium catenulatum subsp. kashiwanohense |
Synonyms | Comment | Organism |
---|---|---|
AH68_00250 | - |
Bifidobacterium catenulatum subsp. kashiwanohense |
BkTauF | - |
Bifidobacterium catenulatum subsp. kashiwanohense |
BwSarD | - |
Bilophila wadsworthia |
sarD | - |
Bifidobacterium catenulatum subsp. kashiwanohense |
sarD | - |
Bilophila wadsworthia |
tauF | - |
Bifidobacterium catenulatum subsp. kashiwanohense |
tauF | - |
Bilophila wadsworthia |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bifidobacterium catenulatum subsp. kashiwanohense |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.8 | - |
NADP+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.97 | - |
isethionate | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.97 | - |
isethionate | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
1.3 | - |
NAD+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
1.3 | - |
NAD+ | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
18.67 | - |
NADH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
18.67 | - |
NADH | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
18.99 | - |
NADPH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
22.83 | - |
2-sulfoacetaldehyde | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
22.83 | - |
2-sulfoacetaldehyde | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
sulfoacetaldehyde reduction | Bifidobacterium catenulatum subsp. kashiwanohense |
10 | - |
assay at | Bifidobacterium catenulatum subsp. kashiwanohense |
10 | - |
isethionate oxidation | Bifidobacterium catenulatum subsp. kashiwanohense |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | NADH/NAD+ is preferred before NADPH/NADP+ by the enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
additional information | enzyme accepts NADP+/NADPH, reaction of EC 1.1.1.313 | Bifidobacterium catenulatum subsp. kashiwanohense | |
NAD+ | - |
Bilophila wadsworthia | |
NAD+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | |
NADH | - |
Bilophila wadsworthia | |
NADH | - |
Bifidobacterium catenulatum subsp. kashiwanohense | |
NADH | preferred cofactor | Bifidobacterium catenulatum subsp. kashiwanohense | |
NADP+ | - |
Bifidobacterium catenulatum subsp. kashiwanohense | |
NADPH | - |
Bifidobacterium catenulatum subsp. kashiwanohense |
General Information | Comment | Organism |
---|---|---|
evolution | sulfoacetaldehyde reductase from the human gut fermenting bacterium Bifidobacterium kashiwanohense (BkTauF) belongs to the M-ADH family, but is distantly related to BwSarD (28% sequence identity), a sulfoacetaldehyde reductase from human gut sulfite-reducing bacterium Bilophila wadsworthia belonging to the metal-dependent alcohol dehydrogenase superfamily (M-ADH). Conservation of active site residues (D192, Q196, F252, T257, H261, F265 and H275) in close homologues of BkTauF | Bifidobacterium catenulatum subsp. kashiwanohense |
evolution | sulfoacetaldehyde reductase SarD, from human gut sulfite-reducing bacterium Bilophila wadsworthia belongs to the metal-dependent alcohol dehydrogenase superfamily (M-ADH). Sulfoacetaldehyde reductase from the human gut fermenting bacterium Bifidobacterium kashiwanohense (BkTauF) belongs to the M-ADH family, but is distantly related to BwSarD (28% sequence identity) | Bilophila wadsworthia |
metabolism | the enzyme is involved in a pathway for taurine dissimilation, in which isethionate is generated as an intermediate, and further degraded to acetate and H2S instead of being secreted | Bilophila wadsworthia |
additional information | the BktauF structure and sequence shows conservation in secondary structure, and metal-coordinating residues except that Gln196 is atypical in the M-ADH family, with His being more common at that position. The putative isethionate-binding site adjacent to the catalytic Zn2+ is very open, which precluded molecular docking, active site structure analysis, overview. The position of isethionate is constrained by the requirements of the M-ADH catalytic mechanism, which requires coordination of the hydroxyl O-atom to Zn2+, and hydride transfer from C1 of isethionate to C4 of the NAD+. Phe252, Thr257 and Phe265 surrounding the active-site cavity are identified as potential substrate-interacting residues | Bifidobacterium catenulatum subsp. kashiwanohense |
physiological function | hydroxyethylsulfonate (isethionate (Ise)) is generated by the sulfoacetaldehyde reductases from human gut bacteria. Isethionate is thought to be derived from aminoethylsulfonate (taurine), as a byproduct of taurine nitrogen assimilation by certain anaerobic bacteria inhabiting the taurine-rich mammalian gut. Sulfoacetaldehyde reductases in Bifidobacteria have a possible role in isethionate production as a byproduct of taurine nitrogen assimilation | Bilophila wadsworthia |
physiological function | hydroxyethylsulfonate (isethionate (Ise)) is generated by the sulfoacetaldehyde reductases from human gut bacteria. Isethionate is thought to be derived from aminoethylsulfonate (taurine), as a byproduct of taurine nitrogen assimilation by certain anaerobic bacteria inhabiting the taurine-rich mammalian gut. Sulfoacetaldehyde reductases in Bifidobacteria have a possible role in isethionate production as a byproduct of taurine nitrogen assimilation | Bifidobacterium catenulatum subsp. kashiwanohense |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.22 | - |
isethionate | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
0.22 | - |
isethionate | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
1.04 | - |
NADP+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
15.8 | - |
NAD+ | pH 10, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
16.25 | - |
NAD+ | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
26.01 | - |
NADPH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
45.66 | - |
2-sulfoacetaldehyde | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense | |
46.1 | - |
2-sulfoacetaldehyde | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
301 | - |
NADH | pH 7.5, temperature not specified in the publication | Bifidobacterium catenulatum subsp. kashiwanohense | |
311.17 | - |
NADH | pH 10.0, 30°C, recombinant wild-type enzyme | Bifidobacterium catenulatum subsp. kashiwanohense |