Recombinant Escherichia coli O157:H7 NADH pyrophosphatase (nudC) is expressed in E. coli and features an N-terminal 6xHis-tag for simplified purification. This full-length protein spans 1-257 amino acids and achieves a purity greater than 85% as verified by SDS-PAGE. This product is designed exclusively for research use and is not intended for therapeutic or diagnostic applications.
NADH pyrophosphatase, encoded by the nudC gene, appears to be involved in regulating NADH levels within the cell. It likely plays a critical role in maintaining cellular redox balance by hydrolyzing NADH, thus influencing various metabolic pathways. Studying this enzyme may be important for understanding bacterial metabolism and energy management, making it a significant focus in microbiological research.
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.
Because nudC is a bacterial enzyme that naturally occurs in E. coli and does not require complex post-translational modifications or eukaryotic chaperones for folding or activity, expression in E. coli is highly likely to yield a properly folded, catalytically active enzyme. Numerous studies have shown that bacterial NADH pyrophosphatases expressed recombinantly in E. coli retain enzymatic activity comparable to the native enzyme. While this recombinant protein is very likely to be correctly folded and enzymatically functional, direct enzymatic validation (e.g., NADH hydrolysis assay) should still be performed to confirm activity before using it in mechanistic or kinetic studies.
1. Biochemical Characterization of NADH Pyrophosphatase Activity
This recombinant nudC protein is well suited for enzymatic assays measuring NADH pyrophosphatase activity, including determination of kinetic parameters (Km, Vmax), substrate specificity, and cofactor dependence. Since the enzyme is expressed full-length in E. coli, which is its native host type, correct folding and catalytic activity are very likely, although confirmation by enzyme activity assay is recommended. The His-tag facilitates purification and quantification, supporting accurate biochemical and kinetic analyses.
2. Comparative Enzyme Studies Between Pathogenic and Non-pathogenic E. coli Strains
This recombinant nudC protein can be used to compare NADH pyrophosphatase activity and structure between E. coli O157:H7 and non-pathogenic strains. Because the enzyme is likely correctly folded and functional, these comparisons can provide reliable data on possible virulence-related or metabolic differences. If activity validation confirms native folding, the protein will serve as a useful reference enzyme for cross-strain biochemical and evolutionary studies.
3. Antibody Development and Immunological Studies
The recombinant protein can be used as an antigen for antibody production (polyclonal or monoclonal) targeting E. coli O157:H7 nudC. Its high purity and bacterial origin minimize immunogenic interference, and the His-tag facilitates downstream assay use (e.g., ELISA or Western blot). The protein’s likely native folding supports antibody generation recognizing both linear and conformational epitopes, though activity confirmation should precede any assumption of structural integrity.
4. Protein-Protein Interaction Studies
The His-tagged full-length nudC can be immobilized on nickel-affinity matrices for pull-down assays to explore potential interactions with other bacterial proteins in NADH metabolism.
Because bacterial expression likely yields a correctly folded enzyme, native-like interactions could be observed. However, interactions should be validated under physiological conditions to ensure biological relevance.
5. Structural Biology and Protein Folding Studies
This recombinant full-length nudC protein (1–257aa) is suitable for X-ray crystallography, NMR, or cryo-EM due to its small size and high purity. Given its bacterial expression and likely proper folding, structural studies are feasible without extensive refolding steps. Biophysical characterization (CD spectroscopy, DSC, DLS) can further confirm structural integrity and thermal stability.
If activity assays confirm functionality, the protein can serve as a model system for catalytic mechanism analysis.
