Recombinant Bacillus subtilis Endoglucanase (eglS) gets expressed in E. coli and includes the complete mature protein sequence, spanning amino acids 30 to 499. The protein carries both an N-terminal 10xHis-tag and a C-terminal V5-tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows it achieves greater than 85% purity, which appears to provide dependable performance for research work.
Endoglucanase from Bacillus subtilis breaks down cellulose by cutting through complex carbohydrates to produce simpler sugars. This enzyme seems central to several biological processes, particularly those related to biomass conversion and energy production. Studying it may help push forward industrial biotechnology research while deepening our grasp of how microbes degrade cellulose.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. Enzyme Kinetics and Biochemical Characterization Studies
This recombinant endoglucanase allows researchers to measure key enzymatic properties like substrate specificity, optimal pH and temperature ranges, plus kinetic parameters including Km and Vmax values. The high purity level (>85%) combined with the dual-tag design makes it well-suited for controlled laboratory assays using different cellulosic substrates. Scientists can build fundamental activity profiles and draw comparisons between this bacterial endoglucanase and cellulolytic enzymes from other species. The N-terminal His-tag makes purification and quantification simpler for standardized kinetic work.
2. Protein-Protein Interaction Studies
Having both tags (N-terminal His-tag and C-terminal V5-tag) opens up multiple experimental routes for protein interaction screening. The His-tag works well for nickel-affinity pull-down experiments to hunt for potential binding partners in bacterial cell lysates or purified protein collections. Meanwhile, the V5-tag supports immunoprecipitation studies with anti-V5 antibodies to confirm interactions discovered through His-tag pull-downs. This dual approach may help cut down false positives in interaction research.
3. Antibody Development and Immunoassay Applications
The recombinant protein makes a solid antigen for creating specific antibodies against Bacillus subtilis endoglucanase using conventional immunization methods. High purity levels suggest minimal contamination that might otherwise cause cross-reactive antibodies. The V5-tag enables sandwich ELISA setups where anti-V5 antibodies function as capture or detection agents alongside newly created anti-endoglucanase antibodies. These immunological tools could then help detect endoglucanase expression in bacterial cultures or environmental samples.
4. Structural Biology and Protein Folding Studies
The purified recombinant protein supplies material for biophysical analysis such as circular dichroism spectroscopy, dynamic light scattering, and thermal stability testing. Since the mature protein region (30-499aa) represents the full functional domain without signal peptide complications, it appears suitable for structural investigations. Scientists can examine protein folding patterns, thermal denaturation behavior, and conformational shifts under various buffer conditions. The His-tag also makes protein immobilization easier for surface plasmon resonance experiments that look at protein stability and conformational changes.
