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Literature summary for 1.14.18.1 extracted from

  • Molloy, S.; Nikodinovic-Runic, J.; Martin, L.B.; Hartmann, H.; Solano, F.; Decker, H.; OConnor, K.E.
    Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches (2013), Biotechnol. Bioeng., 110, 1849-1857.
    View publication on PubMed
Search for more literature for 1.14.18.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Ralstonia solanacearum

Protein Variants

Protein Variants Comment Organism
145-D317Y the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
145-L330V the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
145-V153A the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
145-Y119F the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
C10-F185Y the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
C10-N322S the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
C10-T183I the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
C10-T359M the overall catalytic efficiency for the variant has improved compared to the wild type for D-tyrosine Ralstonia solanacearum
RV145 the random mutation variant exhibits a 3.2fold increase in kcat, respectively, compared to the wild type while the Km is 5.2fold lower. The overall catalytic efficiency for the variant has improved 16fold compared to the wild type for D-tyrosine Ralstonia solanacearum
RVC10 the random mutation variant exhibits a 2.1fold increase in kcat, respectively, compared to the wild type while the Km is 3.2fold lower. The overall catalytic efficiency for the variant has improved 6.7fold compared to the wild type for D-tyrosine Ralstonia solanacearum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
 L-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum
0.9
-
 L-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
1
-
 L-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
1.2
-
 D-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum
1.9
-
 D-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
2.1
-
 L-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
2.4
-
 D-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
3.5
-
 D-tyrosine random mutant enzyme C10-T183I, at pH 7.0 and 30°C Ralstonia solanacearum
5.1
-
 D-tyrosine random mutant enzyme 145-Y119F, at pH 7.0 and 30°C Ralstonia solanacearum
6.2
-
 D-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
6.3
-
 D-tyrosine random mutant enzyme C10-T359M, at pH 7.0 and 30°C Ralstonia solanacearum
6.9
-
 D-tyrosine random mutant enzyme 145-D317Y, at pH 7.0 and 30°C Ralstonia solanacearum
7.3
-
 D-tyrosine random mutant enzyme C10-F185Y, at pH 7.0 and 30°C Ralstonia solanacearum
9.6
-
 D-tyrosine random mutant enzyme 145-L330V, at pH 7.0 and 30°C Ralstonia solanacearum
15
-
 D-tyrosine random mutant enzyme 145-V153A, at pH 7.0 and 30°C Ralstonia solanacearum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
 x * 55000, SDS-PAGE Ralstonia solanacearum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-tyrosine + O2 Ralstonia solanacearum
-
 dopaquinone + H2O
-
 ?
L-tyrosine + O2 Ralstonia solanacearum
-
 dopaquinone + H2O
-
 ?
monophenol + O2 Ralstonia solanacearum
-
 o-diphenol + H2O
-
 ?
o-diphenol + O2 Ralstonia solanacearum
-
 o-quinone + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Ralstonia solanacearum Q8Y2J8
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Ralstonia solanacearum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-tyrosine + O2
-
 Ralstonia solanacearum dopaquinone + H2O
-
 ?
L-tyrosine + O2
-
 Ralstonia solanacearum dopaquinone + H2O
-
 ?
monophenol + O2
-
 Ralstonia solanacearum o-diphenol + H2O
-
 ?
o-diphenol + O2
-
 Ralstonia solanacearum o-quinone + H2O
-
 ?

Subunits

Subunits Comment Organism
? x * 55000, SDS-PAGE Ralstonia solanacearum

Synonyms

Synonyms Comment Organism
catecholase
-
 Ralstonia solanacearum
monophenol monooxygenase
-
 Ralstonia solanacearum
monophenol, L-DOPA: oxygen oxidoreductase
-
 Ralstonia solanacearum
polyphenol oxidase
-
 Ralstonia solanacearum
PPO
-
 Ralstonia solanacearum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
 D-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
0.63
-
 D-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
0.92
-
 L-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum
0.97
-
 D-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum
1.03
-
 L-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
1.22
-
 D-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
1.24
-
 L-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
1.27
-
 D-tyrosine random mutant enzyme 145-Y119F, at pH 7.0 and 30°C Ralstonia solanacearum
1.35
-
 D-tyrosine random mutant enzyme 145-D317Y, at pH 7.0 and 30°C Ralstonia solanacearum
1.37
-
 D-tyrosine random mutant enzyme C10-T183I, at pH 7.0 and 30°C Ralstonia solanacearum
1.37
-
 D-tyrosine random mutant enzyme C10-T359M, at pH 7.0 and 30°C Ralstonia solanacearum
1.42
-
 L-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
1.63
-
 D-tyrosine random mutant enzyme 145-L330V, at pH 7.0 and 30°C Ralstonia solanacearum
1.7
-
 D-tyrosine random mutant enzyme C10-F185Y, at pH 7.0 and 30°C Ralstonia solanacearum
2.1
-
 D-tyrosine random mutant enzyme 145-V153A, at pH 7.0 and 30°C Ralstonia solanacearum

Cofactor

Cofactor Comment Organism Structure
heme
-
 Ralstonia solanacearum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.05
-
 D-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
0.14
-
 D-tyrosine random mutant enzyme 145-V153A, at pH 7.0 and 30°C Ralstonia solanacearum
0.17
-
 D-tyrosine random mutant enzyme 145-L330V, at pH 7.0 and 30°C Ralstonia solanacearum
0.2
-
 D-tyrosine random mutant enzyme 145-D317Y, at pH 7.0 and 30°C Ralstonia solanacearum
0.22
-
 D-tyrosine random mutant enzyme C10-T359M, at pH 7.0 and 30°C Ralstonia solanacearum
0.23
-
 D-tyrosine random mutant enzyme C10-F185Y, at pH 7.0 and 30°C Ralstonia solanacearum
0.25
-
 D-tyrosine random mutant enzyme 145-Y119F, at pH 7.0 and 30°C Ralstonia solanacearum
0.33
-
 D-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
0.39
-
 D-tyrosine random mutant enzyme C10-T183I, at pH 7.0 and 30°C Ralstonia solanacearum
0.51
-
 D-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
0.67
-
 L-tyrosine wild type enzyme, at pH 7.0 and 30°C Ralstonia solanacearum
0.8
-
 D-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum
1.12
-
 L-tyrosine random mutant enzyme RVC10, at pH 7.0 and 30°C Ralstonia solanacearum
1.24
-
 L-tyrosine random mutant enzyme C10-N322S, at pH 7.0 and 30°C Ralstonia solanacearum
1.42
-
 L-tyrosine random mutant enzyme RV145, at pH 7.0 and 30°C Ralstonia solanacearum