Cloned (Comment) | Organism |
---|---|
bsd overexpressed in Escherichia coli BL 21 | Aspergillus terreus |
overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3), Cys91 mutants are expressed in inclusion bodies | Aspergillus terreus |
Crystallization (Comment) | Organism |
---|---|
- |
Aspergillus terreus |
Protein Variants | Comment | Organism |
---|---|---|
C91A | site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure | Aspergillus terreus |
C91H | site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure | Aspergillus terreus |
C91S | site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure | Aspergillus terreus |
E56D | site-directed mutagenesis, inactive mutant | Aspergillus terreus |
E56Q | site-directed mutagenesis, inactive mutant | Aspergillus terreus |
General Stability | Organism |
---|---|
the tetrameric enzyme form is very resistant against denaturation by SDS, 90% remaining activity at 2% SDS, diluted 10fold and incubated 2 h at room temperature | Aspergillus terreus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no inhibition by EDTA and o-phenanthroline | Aspergillus terreus | |
p-Hydroxymercuriphenylsulfonic acid | leads to destabilization of the enzyme structure due to removal of required zinc | Aspergillus terreus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | tightly bound catalytic zinc, 1 zinc ion per subunit, enzyme contains the catalytic zinc-coordinating sequence motif, role in enzyme tetrameric structure stabilization and protein folding, overview | Aspergillus terreus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
13000 | - |
SDS-PAGE, heat treatment | Aspergillus terreus |
13340 | - |
recombinant enzyme, calculated by amino acid residues per subunit | Aspergillus terreus |
36000 | - |
SDS-PAGE without heat treatment | Aspergillus terreus |
49000 | - |
native PAGE | Aspergillus terreus |
49000 | - |
recombinant wild-type enzyme, native PAGE | Aspergillus terreus |
52000 | - |
ultracentrifugal sedimentation analysis | Aspergillus terreus |
54000 | - |
gel filtration | Aspergillus terreus |
54000 | - |
recombinant and native wild-type enzyme, gel filtration | Aspergillus terreus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
blasticidin S + H2O | Bacillus cereus | - |
deaminohydroxyblasticidin S + NH3 | - |
? | |
blasticidin S + H2O | Aspergillus terreus | - |
deaminohydroxyblasticidin S + NH3 | - |
? |
Organic Solvent | Comment | Organism |
---|---|---|
SDS | quite resistant to denaturation | Aspergillus terreus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus terreus | - |
- |
- |
Bacillus cereus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Aspergillus terreus |
native enzyme from Aspergiluus terreus, recombinant His-tagged enzyme from Escherichia coli by nickel chelate affinity chromatography | Aspergillus terreus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
blasticidin S + H2O = deaminohydroxyblasticidin S + NH3 | catalyses the deamination of the cytosine moiety of the antibiotics blasticidin S, cytomycin and acetylblasticidin S, Glu56 is required for catalysis, and Cys91 is required for structural maintenance | Aspergillus terreus |
Renatured (Comment) | Organism |
---|---|
reconstitution of the enzyme after denaturation with guanidine-HCl, acid, or p-hydroxymercuriphenylsulfonic acid, using different divalent metal ions, renaturation of the enzyme is completely blocked in presence of 1 mM EDTA, refolding of recombinant Cys91 mutant enzymes from inclusion bodies | Aspergillus terreus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
blasticidin S + H2O | - |
Bacillus cereus | deaminohydroxyblasticidin S + NH3 | - |
? | |
blasticidin S + H2O | - |
Aspergillus terreus | deaminohydroxyblasticidin S + NH3 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | zinc has a role in enzyme tetrameric structure stabilization and protein folding | Aspergillus terreus |
tetramer | 4 * 13000, SDS-PAGE | Aspergillus terreus |
tetramer | 4 * 13000, SDS-PAGE containing 15% SDS | Aspergillus terreus |
Synonyms | Comment | Organism |
---|---|---|
blasticidin S deaminase | - |
Aspergillus terreus |
BSD | - |
Aspergillus terreus |