BirA Biotin-protein Ligase

Description
Biotin-protein ligase (EC 6.3.4.15) activates biotin to form biotinyl 5' adenylate and transfers the biotin to biotin-accepting proteins. It also functions as a biotin operon repressor. The protein is encoded by the birA gene. Other names for this enzyme include: biotin ligase; biotin operon repressor protein; birA; biotin holoenzyme synthetase; biotin-(acetyl-CoA carboxylase) synthetase.

Applications

Western blot

Functional Studies of enzyme kinetics

Screening inhibitors and selectivity profiling

Source
E. coli

Molecule Weight
The recombinant Biotin-protein ligase consists of amino acids 2-321 and has a predicted molecular mass of 37 kDa.

Purity
> 95 % as determined by SDS-PAGE

Activity

 5,000 Units/µg

Definition of Activity
1 Unit is the amount of enzyme that will biotinylate 1 pmol of peptide substrate in 30 minutes at 30°C using the reaction buffers provided and 40 µM peptide substrate*.
*The peptide substrate used in the enzyme assays was a 27kDa AviTag'd recombinant protein expressed by Pichia pastoris.

Contaminating proteases: <0.01% as chymotrypsin-like activity

Storage: -80 Avoid repeated freeze-thaw cycles.

Storage Buffer
50 mM Tris-HCl, pH 7.5, 50 mM NaCl, 1 mM DTT, and 50% Glycerol

Shipping: Dry ice