Application | Comment | Organism |
---|---|---|
synthesis | the enzyme can be useful in the production of retinoic acid and related high-end products | Bacillus cereus |
Cloned (Comment) | Organism |
---|---|
recombinant tagged enzyme expression in Escherichia coli strain ER2566 | Bacillus cereus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant BcALDH alone and in complex with NAD+ and NADP+ cofactors, sitting drop vapour diffusion method, mixing of 10 mg/ml protein in 25 mM Tris, pH 7.5, 15 mM NaCl, and 3 mM 2-mercaptoethanol, with crystallization solution containing 0.15 M DL-malic acid, pH 7.0, and 18% PEG 3350 w/v, at 14°C, 1 week, X-ray diffraction structure determination and analysis at 2.0-2.6 A resolution, molecular replacement using human ALDH2 structure (PDB ID 4FR8) as the search model, modeling. The apo-BcALDH structure contains four protomers in the asymmetric unit | Bacillus cereus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus cereus | A0A150BLG9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant tagged enzyme from Escherichia coli strain ER2566 by metal chelating affinity chromatography, desalting gel filtration, and dialysis | Bacillus cereus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetaldehyde + NAD(P)+ + H2O | - |
Bacillus cereus | acetate + NAD(P)H + H+ | - |
? | |
all-trans-retinal + NAD+ | - |
Bacillus cereus | all-trans-retinol + NADH + H+ | - |
? | |
all-trans-retinal + NADP+ | - |
Bacillus cereus | all-trans-retinol + NADPH + H+ | - |
? | |
all-trans-retinol + NAD+ + H2O | - |
Bacillus cereus | all-trans-retinoic acid + NADH + H+ | - |
? | |
all-trans-retinol + NADP+ + H2O | - |
Bacillus cereus | all-trans-retinoic acid + NADPH + H+ | - |
? | |
benzaldehyde + NAD(P)+ + H2O | - |
Bacillus cereus | benzoate + NAD(P)H + H+ | - |
? | |
additional information | in the first acylation step, the activated catalytic Cys, Cys300 in BcALDH, attacks the substrate aldehyde group to form a thio-hemiacetal intermediate. The oxidized NAD(P)+ cofactor takes up a hydride ion from the thio-hemiacetal intermediate, which transforms it to a thioester intermediate. This intermediate is then deacylated by a water molecule, in which the additional catalytic residue Glu, Glu266 in BcALDH, activates the hydrolytic water molecule by abstracting a proton. Finally, the reduced NAD(P)H is released. Plausible mechanism for the minor reducing activity of BcALDH with all-trans-retinal, overview. The beta-ionone ring of the superimposed retinoic acid (REA) is bound at the mouth of the channel and exposed to the solvent. The substrate binding pocket-bound REA shows a bent conformation in the carbon positions from 9 to 11. Molecular docking of retinoic acid into the enzyme structure. BcALDH has a strict substrate preference for oxidation of all-trans-retinal, with no activity detected against 9-cis-retinal and 13-cis-retinal. BcALDH also has 14 and 30fold lower Km values for all-trans-retinal compared to the small aldehydes acetaldehyde and benzaldehyde, respectively | Bacillus cereus | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
BcALDH | - |
Bacillus cereus |
More | see also EC 1.2.1.36 | Bacillus cereus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the nicotinamide moieties of NAD+ and NADP+ directly interact with the conserved catalytic residues Cys300 and Glu266, and binding causes concerted conformational changes. Flexible cofactor-binding pocket of BcALDH. Cofactor-enzyme binding structure analysis, overview | Bacillus cereus | |
NAD+ | - |
Bacillus cereus | |
NADP+ | - |
Bacillus cereus |
General Information | Comment | Organism |
---|---|---|
additional information | enzyme structure comparisons, overview | Bacillus cereus |
physiological function | enzyme BcALDH is an atypical aldehyde dehydrogenase having bidirectional oxidizing and reducing activities | Bacillus cereus |