Recombinant Bifunctional ligase/repressor BirA, His (rBirA,His)
Squence
Amino Acid Sequence
Synonyms
Accession Number
GeneID
Summary
BirA, the biotin-protein ligase (BPL) of Escherichia coli, is also known as biotin operon repressor, biotin-[acetyl-CoA-carboxylase] ligase, and biotin-[acetyl-CoAcarboxylase] synthetase. BirA, a member of the group II biotin-protein ligase family, contains an N-terminal helix-turn-helix DNA-binding domain, a catalytic core that catalyzes biotinyl 5′ adenylate (bio-5′-AMP) synthesis, and a C-terminal domain that plays a role in DNA binding, dimerization, and catalytic function. BirA functions both as a DNA-binding protein that represses the biotin biosynthesis operon as well as an enzyme that synthesizes its own corepressor, bio-5′-AMP, an intermediate in biotinylation reactions. BirA biotinylates via the lysine side chain of biotin-accepting proteins/peptides, including natural substrate, carboxyl carrier protein (BCCP),and Avi Tag fusion proteins. Once biotinylated, (strept)avidin-biotin interactions can be utilized in a wide variety of applications of biochemistry and cell biology, including protein capture, immobilization, multimerizing, and bridging molecules.
Source
Escherichia coli.
Molecular Weight
Biological Activity
Measured by its ability to generate pyrophosphate from the biotinylation reaction. The pyrophosphate is subsequently hydrolyzed using Recombinant Yeast Inorganic Pyrophosphatase/PPA1 (ryPPA1). The specific activity is > 10.0 pmol/min/ug, as measured under the described conditions.
Appearance
Sterile colorless liquid.
Formulation
Supplied as a 0.2 um filtered solution in 50 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM DTT, 10 % glycerol.
Endotoxin
Less than 0.1 EU/ug of rBirA, His as determined by LAL method.
Reconstitution
Stability and Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 6 months from date of receipt, -20 to -70 °C as supplied.- 3 months, -20 to -70 °C under sterile conditions after opening.
