Any feedback?
Literature summary extracted from
- An asymmetric reductase that intercepts acyclic imino acids produced in situ by a partner oxidase (2019), J. Am. Chem. Soc., 141, 12258-12267 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.1.1.B66 | gene bsp5, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant overexpression of His6-tagged wild-type and mutant enzymes from pUC57-Bsp5 vector in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Bacillus sp. 5mfcol3.1 |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 1.1.1.B66 | purified enzyme Bsp5 in complex with D-arginine and coenzyme NADPH, hanging drop vapour diffusion, mixing of 800 nl of 10 mg/m protein in 20 mM HEPES, 50 mM NaCl, pH 7.5, 5 mM D-Arg, and 5 mM NADPH, with 800 nl of reservoir solution containing 20% w/v PEG 3350 and 0.2 M sodium tartrate dibasic, equilibration against 0.05 ml of reservoir solution, room temperature, 10 days, method optimization, X-ray diffraction structure determination and analysis at 1.6 A resolution, molecular replacement, modeling | Bacillus sp. 5mfcol3.1 |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.1.1.B66 | H130A | site-directed mutagenesis, the mutant shows highly reduced activity with substrate (4E)-5-carbamimidamido-2-iminopent-4-enoic acid, but unaltered activity with 5-carbamimidamido-2-iminopentanoic acid compared to wild-type | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | H130A/Y132F | site-directed mutagenesis, the mutant shows highly reduced activity with substrate (4E)-5-carbamimidamido-2-iminopent-4-enoic acid, but increased activity with 5-carbamimidamido-2-iminopentanoic acid compared to wild-type | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | additional information | the mutations do not impact dimerization | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | R229A | site-directed mutagenesis, the mutant shows reduced activity with substrates (4E)-5-carbamimidamido-2-iminopent-4-enoic acid and 5-carbamimidamido-2-iminopentanoic acid compared to wild-type | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | Y132F | site-directed mutagenesis, the mutant shows highly reduced activity with substrate (4E)-5-carbamimidamido-2-iminopent-4-enoic acid, but unaltered activity with 5-carbamimidamido-2-iminopentanoic acid compared to wild-type | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | Y97F | site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme | Bacillus sp. 5mfcol3.1 |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.B66 | 70000 | - |
about, recombinant His-tagged enzyme, gel filtration | Bacillus sp. 5mfcol3.1 |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.B66 | additional information | Bacillus sp. 5mfcol3.1 | enzyme Bsp5 reduces acyclic imino acids produced in situ by a partner oxidase | ? | - |
- |
 |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.1.1.B66 | Bacillus sp. 5mfcol3.1 | A0A1I4FUG4 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.1.1.B66 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration | Bacillus sp. 5mfcol3.1 |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.1.1.B66 | an acyclic iminoacid + NADPH + H+ = an amino acid + NADP+ | no imine reducing activity on cyclic compounds | Bacillus sp. 5mfcol3.1 |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.B66 | (4E)-5-carbamimidamido-2-iminopent-4-enoic acid + NADPH + H+ | - |
Bacillus sp. 5mfcol3.1 | D-2-dehydroarginine + NADP+ | - |
? | |
| 1.1.1.B66 | 5-carbamimidamido-2-iminopentanoic acid + NADPH + H+ | Bsp5 shows poor activity in the reverse reaction direction | Bacillus sp. 5mfcol3.1 | D-arginine + NADP+ | - |
? | |
| 1.1.1.B66 | additional information | enzyme Bsp5 reduces acyclic imino acids produced in situ by a partner oxidase | Bacillus sp. 5mfcol3.1 | ? | - |
- |
|
| 1.1.1.B66 | additional information | the asymmetric reductase intercepts acyclic imino acids produced in situ by a partner oxidase (Ind4 from Paenibacillus sp.) in a coupled assay, overview. Nonenzymatic hydrolysis of acyclic imino acids gives compounds (4E)-5-carbamimidamido-2-oxopent-4-enoic acid and 5-carbamimidamido-2-oxopentanoic acid. Bsp5 fails to reduce 5-carbamimidamido-2-oxopentanoic acid, and incubation of Bsp5 with 2-methylpyrroline also results in no reaction. Bsp5 lacks imine reducing activity on cyclic substrates. LC-MS analysis of the molecules | Bacillus sp. 5mfcol3.1 | ? | - |
- |
|
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.1.1.B66 | homodimer | 2 * 35939, recombinant wild-type enzyme, sequence calculation and mass spectrometry | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | More | dimer three-dimensional structure modeling with dimeric interface, overview | Bacillus sp. 5mfcol3.1 |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.1.1.B66 | asymmetric reductase | - |
Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | BSp5 | - |
Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | imino acid reductase | - |
Bacillus sp. 5mfcol3.1 |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.B66 | 30 | - |
assay at | Bacillus sp. 5mfcol3.1 |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.B66 | 7.5 | - |
assay at | Bacillus sp. 5mfcol3.1 |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.1.B66 | NADP+ | - |
Bacillus sp. 5mfcol3.1 | |
| 1.1.1.B66 | NADPH | - |
Bacillus sp. 5mfcol3.1 | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.1.1.B66 | evolution | enzyme Bsp5 belongs to the D-2-hydroxyacid dehydrogenase family | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | additional information | structure-based mechanism of Bsp5's imine reductase activity | Bacillus sp. 5mfcol3.1 |
| 1.1.1.B66 | physiological function | Bsp5 is an imino acid reductase from the D-2-hydroxyacid dehydrogenase family that reduces acyclic imino acids produced in situ by a partner oxidase | Bacillus sp. 5mfcol3.1 |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
