Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Ovchinnikova, M.V.; Mikhailova, A.G.; Karlinsky, D.M.; Gorlenko, V.A.; Rumsh, L.D.
    Reversible cyclic thermal inactivation of oligopeptidase B from Serratia proteamaculans (2018), Acta Naturae, 10, 65-70 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.83 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Serratia proteamaculans

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.83 D647A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 2fold lower Serratia proteamaculans
3.4.21.83 D649A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
3.4.21.83 E125A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 20-25% higher than that of the wild-type enzyme Serratia proteamaculans
3.4.21.83 E75A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 20-25% higher than that of the wild-type enzyme Serratia proteamaculans
3.4.21.83 H652X site-directed mutagenesis Serratia proteamaculans
3.4.21.83 K655A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
3.4.21.83 K660A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
3.4.21.83 additional information calcixadum ions are a destabilizing factor for all PSP mutant variants. At 50 mM Ca2+, the difference in the thermal stability of all PSP variants is less marked, but the E75A mutant is the most thermally labile one, detailed overview. Substitution of the corresponding charged amino acid residues for the unxadcharged ones in OpdB from Trypanosoma brucei significantly rexadduces the thermal stability of these mutants Serratia proteamaculans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.21.83 Ca2+ calcium ions impede the reverse transition to the closed form. The reduction in the thermal stability of PSP in the presence of Ca2+ can be attributed to the destruction of the salt bridges SB2 and SB3 that takes place as Ca2+ binds to the E494 and D460 residues parxadtaking in bridge formation. Calcixadum ions are a destabilizing factor for all PSP mutant variants, overview Serratia proteamaculans

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.83 Serratia proteamaculans A8GHH5
-
-
3.4.21.83 Serratia proteamaculans 568 A8GHH5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.83 recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Serratia proteamaculans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.83 Nalpha-benzoyl-DL-argininyl-4-nitroanilide + H2O i.e. BAPNA, substrate of wild-type enzyme PSP, lower activity with truncated enzyme PSP-Chtr Serratia proteamaculans Nalpha-benzoyl-DL-arginine + 4-nitroaniline
-
 ?
3.4.21.83 Nalpha-benzoyl-DL-argininyl-4-nitroanilide + H2O i.e. BAPNA, substrate of wild-type enzyme PSP, lower activity with truncated enzyme PSP-Chtr Serratia proteamaculans 568 Nalpha-benzoyl-DL-arginine + 4-nitroaniline
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.21.83 OpdB
-
 Serratia proteamaculans
3.4.21.83 PSP
-
 Serratia proteamaculans
3.4.21.83 Spro_3467
-
 Serratia proteamaculans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.21.83 25
-
 assay at Serratia proteamaculans

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.21.83 37
-
 wild-type and mutant PSP enzymes can undergo reversible thermal inactivation at 37°C, pH 8.0, with activity being restored or even increased with respect to the initial one upon subsequent cooling. The process can be repeated several times, with the same results achieved (up to 5 cycles). This effect can be explained by a shift in the equilibrium between the inactive open form of the enzyme and the active closed one upon variation of the incubation temperature, overview Serratia proteamaculans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.21.83 8
-
 assay at Serratia proteamaculans

General Information

EC Number General Information Comment Organism
3.4.21.83 evolution oligopeptidase B (OpdB) is a trypsin-like serine peptidase belonging to the prolyl oligopeptidase family Serratia proteamaculans
3.4.21.83 malfunction substitution of the corresponding charged amino acid residues for the unxadcharged ones in OpdB from Trypanosoma brucei significantly rexadduces the thermal stability of these mutants Serratia proteamaculans