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Literature summary for 1.4.3.10 extracted from

  • Kopacz, M.M.; Rovida, S.; van Duijn, E.; Fraaije, M.W.; Mattevi, A.
    Structure-based redesign of cofactor binding in putrescine oxidase (2011), Biochemistry, 50, 4209-4217.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Top10 cells Rhodococcus erythropolis

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 2.0-2.4 M ammonium sulfate, 0.1 M sodium citrate, and 0.1 M MES-HCl pH 6.4 Rhodococcus erythropolis

Protein Variants

Protein Variants Comment Organism
A394C the mutant enzyme does not contain covalently bound flavin Rhodococcus erythropolis
A394C/A396T/Q431G mutant with strongly reduced catalytic efficiency Rhodococcus erythropolis
A394C/V433M the mutations improve the affinity toward FAD Rhodococcus erythropolis
P15I/A394C the mutations improve the affinity toward FAD Rhodococcus erythropolis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0134
-
putrescine mutant enzyme P15I/A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
0.0197
-
putrescine mutant enzyme A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
0.021
-
putrescine mutant enzyme A394C/A396T/Q431G, at pH 8.3 and 25°C Rhodococcus erythropolis
0.0454
-
putrescine wild type enzyme, at pH 8.3 and 25°C Rhodococcus erythropolis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
putrescine + O2 + H2O Rhodococcus erythropolis
-
4-aminobutanal + NH3 + H2O2
-
?

Organism

Organism UniProt Comment Textmining
Rhodococcus erythropolis B0F9F6
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Rhodococcus erythropolis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
putrescine + O2 + H2O
-
Rhodococcus erythropolis 4-aminobutanal + NH3 + H2O2
-
?

Subunits

Subunits Comment Organism
homodimer
-
Rhodococcus erythropolis

Synonyms

Synonyms Comment Organism
PuO
-
Rhodococcus erythropolis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.79
-
putrescine mutant enzyme P15I/A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
0.95
-
putrescine mutant enzyme A394C/A396T/Q431G, at pH 8.3 and 25°C Rhodococcus erythropolis
2.4
-
putrescine mutant enzyme A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
19
-
putrescine wild type enzyme, at pH 8.3 and 25°C Rhodococcus erythropolis

Cofactor

Cofactor Comment Organism Structure
ADP ADP competes with FAD, about 50% of wild type monomers isolated are occupied by ADP instead of FAD Rhodococcus erythropolis
FAD
-
Rhodococcus erythropolis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
45
-
putrescine mutant enzyme A394C/A396T/Q431G, at pH 8.3 and 25°C Rhodococcus erythropolis
59
-
putrescine mutant enzyme P15I/A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
123
-
putrescine mutant enzyme A394C, at pH 8.3 and 25°C Rhodococcus erythropolis
419
-
putrescine wild type enzyme, at pH 8.3 and 25°C Rhodococcus erythropolis