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Literature summary for 6.3.4.15 extracted from

  • Parthasarathy, R.; Bajaj, J.; Boder, E.T.
    An immobilized biotin ligase: surface display of Escherichia coli BirA on Saccharomyces cerevisiae (2005), Biotechnol. Prog., 21, 1627-1631.
    View publication on PubMed

Application

Application Comment Organism
synthesis the enzyme is utilized to synthesize site-specific biotinylated proteins via a biotin acceptor peptide tag Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
gene birA, functional expression of N-terminally and C-terminally multiply tagged enzyme in Saccharomyces cerevisiae, low level expression of the surface-displayed enzyme, expression of avitagged-BirA in Saccharomyces cerevisiae, overview Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
glycerol inhibits the recombinant enzyme at 10% Escherichia coli
NaCl inhibits the purified enzyme at 0.1 M, but not the immobilized enzyme in crude extract of recombinant yeast cells Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0033
-
avitag-tagged maltose-binding protein recombinant enzyme Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
x * 35000, recombinant enzyme, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
avitag-tagged maltose-binding protein + biotin + ATP Escherichia coli recombinant Saccharomyces cerevisiae cells maltose-binding protein-avitag-biotin + AMP + diphosphate
-
?
avitagged BirA + biotin + ATP Escherichia coli recombinant Saccharomyces cerevisiae cells BirA-avitag-biotin + AMP + diphosphate
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Saccharomyces cerevisiae, immobilization of the recombinant enzyme Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
the immobilized enzyme is slightly less active than the soluble enzyme Escherichia coli

Storage Stability

Storage Stability Organism
4°C, immobilized recombinant enzyme, PBS, completely stable for 4 weeks, loss of activity after 5 months Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
avitag-tagged maltose-binding protein + biotin + ATP recombinant Saccharomyces cerevisiae cells Escherichia coli maltose-binding protein-avitag-biotin + AMP + diphosphate
-
?
avitag-tagged maltose-binding protein + biotin + ATP recombinant avitag-tagged maltose-binding protein Escherichia coli maltose-binding protein-avitag-biotin + AMP + diphosphate
-
?
avitagged BirA + biotin + ATP recombinant Saccharomyces cerevisiae cells Escherichia coli BirA-avitag-biotin + AMP + diphosphate
-
?
avitagged BirA + biotin + biotin + ATP the recombinant avitagged enzyme biotinylates itself at the avitag sequence Escherichia coli BirA-avitag-biotin + AMP + diphosphate
-
?
additional information the enzyme site-specifically biotinylates proteins via a biotin acceptor peptide tag Escherichia coli ?
-
?

Subunits

Subunits Comment Organism
? x * 35000, recombinant enzyme, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
biotin ligase
-
Escherichia coli
BirA
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli