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Literature summary for 6.3.4.15 extracted from

  • Lu, W.; Levy, M.; Kincaid, R.; Ellington, A.
    Directed evolution of the substrate specificity of biotin ligase (2014), Biotechnol. Bioeng., 111, 1071-1081 .
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
I11V/P85S/ M157T/K163E/I224T 17fold increase in the ability to use desthiobiotin as a substrate Escherichia coli
M157T 2-3fold increase in kcat value for desthiobiotin as a substrate Escherichia coli
Q41R/M157T/I224T/E215D/T225A 10fold increase in the ability to use desthiobiotin as a substrate Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0017
-
desthiobiotin wild-type, pH 8.3, 30°C Escherichia coli
0.003
-
biotin wild-type, pH 8.3, 30°C Escherichia coli
0.0043
-
desthiobiotin mutant M157T, pH 8.3, 30°C Escherichia coli
0.0067
-
desthiobiotin mutant I11V/P85S/ M157T/K163E/I224T, pH 8.3, 30°C Escherichia coli
0.22
-
ATP wild-type, pH 8.3, 30°C Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + biotin + CGGGSGGGSGLNDIFEAQKIEWH
-
Escherichia coli AMP + diphosphate + biotinylated CGGGSGGGSGLNDIFEAQKIEWH
-
?
ATP + desthiobiotin + CGGGSGGGSGLNDIFEAQKIEWH
-
Escherichia coli AMP + diphosphate + desthiobiotinylated CGGGSGGGSGLNDIFEAQKIEWH
-
?

Synonyms

Synonyms Comment Organism
BirA
-
Escherichia coli
BPL
-
Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.29
-
desthiobiotin wild-type, pH 8.3, 30°C Escherichia coli
0.3
-
biotin wild-type, pH 8.3, 30°C Escherichia coli
0.48
-
ATP wild-type, pH 8.3, 30°C Escherichia coli
0.53
-
desthiobiotin mutant M157T, pH 8.3, 30°C Escherichia coli
0.73
-
desthiobiotin mutant I11V/P85S/ M157T/K163E/I224T, pH 8.3, 30°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
100
-
desthiobiotin mutant I11V/P85S/ M157T/K163E/I224T, pH 8.3, 30°C Escherichia coli
120
-
desthiobiotin mutant M157T, pH 8.3, 30°C Escherichia coli
170
-
desthiobiotin wild-type, pH 8.3, 30°C Escherichia coli
1050
-
biotin wild-type, pH 8.3, 30°C Escherichia coli