Purification and Quality Control
The His-tag full length recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified BirA is greater than 95% homogeneous based on SDS-PAGE analysis.

Unit Definition (Activity)
1 ug is sufficient to transfer biotin into 50ug target protein at room temperature or at 4°C overnight (>95%).

Applications
As soluble active recombinant protein, may be used for specifically transfer a biotin group to a specific lysine (K) on target protein which possessing a target amino acid sequence, such as LHHILDAQKMVWNHR in vitro. For Research use only.

Formulation and Storage
The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles.

Synonym
Bifunctional protein BirA, Biotin--protein ligase

Protein Sequence
MKDNTVPLKL IALLANGEFH SGEQLGETLG MSRAAINKHI QTLRDWGVDV FTVPGKGYSL
PEPIQLLNAK QILGQLDGGS VAVLPVIDST NQYLLDRIGE LKSGDACIAE YQQAGRGRRG
RKWFSPFGAN LYLSMFWRLE QGPAAAIGLS LVIGIVMAEV LRKLGADKVR VKWPNDLYLQ
DRKLAGILVE LTGKTGDAAQ IVIGAGINMA MRRVEESVVN QGWITLQEAG INLDRNTLAA
MLIRELRAAL ELFEQEGLAP YLSRWEKLDN FINRPVKLII GDKEIFGISR GIDKQGALLL
ERME

Background
BirA acts both as a biotin-operon repressor and as the enzyme that synthesizes the corepressor, acetyl-CoA:carbon-dioxide ligase. This protein also activates biotin to form biotinyl-5'-adenylate and transfers the biotin moiety to biotin-accepting proteins.
ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)] = AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)].

References
1. Howard P.K., Shaw J., Otsuka A.J. (1985) Gene 35:321-331.
2. Wilson K.P., Shewchuk L.M., Brennan R.G., Otsuka A.J., Matthews B.W., (1992) Proc. Natl. Acad. Sci. U.S.A. 89:9257-9261

DISCLAIMER

This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.