Recombinant Bifunctional ligase/repressor BirA
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. |
| M.Wt | Approximately 35.3 kDa, a single non-glycosylated polypeptide chain containing 321 amino acids. |
| AA Sequence | MKDNTVPLKL IALLANGEFH SGEQLGETLG MSRAAINKHI QTLRDWGVDV FTVPGKGYSL PEPIQLLNAK QILGQLDGGS VAVLPVIDST NQYLLDRIGE LKSGDACIAE YQQAGRGRRG RKWFSPFGAN LYLSMFWRLE QGPAAAIGLS LVIGIVMAEV LRKLGADKVR VKWPNDLYLQ DRKLAGILVE LTGKTGDAAQ IVIGAGINMA MRRVEESVVN Q |
| Appearance | Sterile colorless liquid. |
| Stability & 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. |
| Formulation | Supplied as a 0.2 μm filtered solution in 50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 10 % glycerol. |
| 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/μg, as measured under the described conditions. |
| Shipping Condition | Blue ice |
| Handling | Centrifuge the vial prior to opening. |
| Usage | For Research Use Only! Not to be used in humans. |
Quality Control & DataSheet
- View current batch:
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Purity > 97 % by SDS-PAGE analyses.
- Datasheet
Endotoxin: Less than 0.1 EU/μg of rBirA as determined by LAL method.