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Literature summary extracted from

  • Kimura, M.; Sekido, S.; Isogai, Y.; Yamaguchi, I.
    Expression, purification, and characterization of blasticidin S deaminase (BSD) from Aspergillus terreus: The role of catalytic zinc in enzyme structure (2000), J. Biochem., 127, 955-963.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.4.23 bsd overexpressed in Escherichia coli BL 21 Aspergillus terreus
3.5.4.23 overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3), Cys91 mutants are expressed in inclusion bodies Aspergillus terreus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.4.23
-
 Aspergillus terreus

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.23 C91A site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure Aspergillus terreus
3.5.4.23 C91H site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure Aspergillus terreus
3.5.4.23 C91S site-directed mutagenesis, inactive mutant, gross perturbation of the enzyme structure Aspergillus terreus
3.5.4.23 E56D site-directed mutagenesis, inactive mutant Aspergillus terreus
3.5.4.23 E56Q site-directed mutagenesis, inactive mutant Aspergillus terreus

General Stability

EC Number General Stability Organism
3.5.4.23 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

EC Number Inhibitors Comment Organism Structure
3.5.4.23 additional information no inhibition by EDTA and o-phenanthroline Aspergillus terreus
3.5.4.23 p-Hydroxymercuriphenylsulfonic acid leads to destabilization of the enzyme structure due to removal of required zinc Aspergillus terreus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.4.23 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.4.23 13000
-
 SDS-PAGE, heat treatment Aspergillus terreus
3.5.4.23 13340
-
 recombinant enzyme, calculated by amino acid residues per subunit Aspergillus terreus
3.5.4.23 36000
-
 SDS-PAGE without heat treatment Aspergillus terreus
3.5.4.23 49000
-
 native PAGE Aspergillus terreus
3.5.4.23 49000
-
 recombinant wild-type enzyme, native PAGE Aspergillus terreus
3.5.4.23 52000
-
 ultracentrifugal sedimentation analysis Aspergillus terreus
3.5.4.23 54000
-
 gel filtration Aspergillus terreus
3.5.4.23 54000
-
 recombinant and native wild-type enzyme, gel filtration Aspergillus terreus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.4.23 blasticidin S + H2O Bacillus cereus
-
 deaminohydroxyblasticidin S + NH3
-
 ?
3.5.4.23 blasticidin S + H2O Aspergillus terreus
-
 deaminohydroxyblasticidin S + NH3
-
 ?

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.5.4.23 SDS quite resistant to denaturation Aspergillus terreus

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.23 Aspergillus terreus
-
-
-
3.5.4.23 Bacillus cereus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.23
-
 Aspergillus terreus
3.5.4.23 native enzyme from Aspergiluus terreus, recombinant His-tagged enzyme from Escherichia coli by nickel chelate affinity chromatography Aspergillus terreus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.4.23 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
a

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.5.4.23 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.23 blasticidin S + H2O
-
 Bacillus cereus deaminohydroxyblasticidin S + NH3
-
 ?
3.5.4.23 blasticidin S + H2O
-
 Aspergillus terreus deaminohydroxyblasticidin S + NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.4.23 More zinc has a role in enzyme tetrameric structure stabilization and protein folding Aspergillus terreus
3.5.4.23 tetramer 4 * 13000, SDS-PAGE Aspergillus terreus
3.5.4.23 tetramer 4 * 13000, SDS-PAGE containing 15% SDS Aspergillus terreus

Synonyms

EC Number Synonyms Comment Organism
3.5.4.23 blasticidin S deaminase
-
 Aspergillus terreus
3.5.4.23 BSD
-
 Aspergillus terreus