In Vivo Enzymatic Biotinylation of Proteins

With the rapid development of biotechnology, biotinylation has become an important research tool in proteomics. Biotin is a naturally occurring metabolic enzyme factor that exhibits activity through the action of specific protein-biotin ligases. The physiological functions of organisms are mainly controlled and regulated by proteins in cells. Using efficient hands-free co-precipitation technique is one of the typical methods to detect protein interaction. Biotin is a soluble small molecule produced by plants or prokaryotes, which can be non-covalently combined with avidin or streptavidin, making the biotin-avidin system widely used in the field of biotechnology middle. The biotinylation of the target protein can be achieved through a biotinylation tag. Avi tag is a short peptide tag consisting of 15 amino acid residues, which can be biotinylated by biotin ligase (BirA enzyme) in vivo or in vitro. Alfa Chemistry aims to construct an efficient expression vector, so as to establish an in vivo enzymatic biotinylation system, so that the target protein is biotinylated.

The Services

Alfa Chemistry provides a professional protein biotinylation system in vivo, which promotes more efficient enzymatic protein biotinylation in vivo. The services we can provide include but are not limited to:

• Efficient enzymatic biotinylation in vivo
• Biotinylation solutions and strategies
• Professional data analysis
• Efficient detection of protein interaction

About biotinylation and biotin ligase

Biotinylation is widely used in biotechnology, biochemistry, immunobiology and cell biology. Biotin is an essential coenzyme that can be synthesized by plants, most bacteria and fungi and participates in several important physiological processes. Biotin becomes biologically active when the protein and biotin are covalently linked. Biotin protein ligase (BPL), which can catalyze the efficient biotinylation of biotin and protein. Every organism has a single BPL-encoding gene. Biotin-dependent enzymes are ubiquitous in nature, biotinylation is a rare modification, and only a few types of biotinylated proteins have been found in organisms. BPL can carry out efficient and specific biotinylation reaction. The most common biotin ligase is the multifunctional BirA enzyme in E. coli.

The advantages

• High sensitivity: There is a strong interaction between biotin and streptavidin, which enables efficient and stable capture of proteins and complexes
• High specificity: There is a highly specific binding between biotin-streptavidin
• High adaptability: Biotinylated proteins can be purified efficiently

Interaction of biotin ligase with its protein substrate

Enzyme carriers containing biotin are usually located at the C-terminus of carboxylases, with biotin-lysine approximately 35 residues from the C-terminus. Biotinylation of a biotin carrier protein requires at least 75-80 residues, with approximately 35-40 residues flanking the biotin site for biotinylation.

Alfa Chemistry provides high-quality biotinylation solutions and strategies to meet the requirements for efficient in vivo protein biotinylation. If you are interested in our services, please contact us immediately.

Reference:

1. Anne. C.S.; et al. In vivo enzymatic protein biotinylation. Biomolecular Engineering. 1999, 16(1-4): 119-125.

※ It should be noted that our service is only used for research, not for clinical use.