Recombinant Citrus unshiu Limonoid UDP-glucosyltransferase is produced through a baculovirus expression system, which appears to deliver solid fidelity and consistency. The complete protein covers amino acids 1-511 and includes an N-terminal 10xHis tag plus a C-terminal Myc tag - both designed to make purification and detection more straightforward. SDS-PAGE analysis indicates the product reaches purity levels above 85%, suggesting it should work well for different research applications.
Limonoid UDP-glucosyltransferase is an enzyme that handles the glucosylation of limonoids, which are secondary metabolites commonly found in citrus plants. This enzyme may play a significant role in modifying these limonoids, likely affecting both their solubility and bioactivity. It represents an important target for studies examining plant biochemistry and efforts to improve citrus fruit quality.
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.
Citrus unshiu Limonoid UDP-glucosyltransferase is a plant enzyme that requires precise folding, proper UDP-sugar binding site formation, and specific tertiary structure for its functional activity in glycosyl transfer. The baculovirus expression system provides a eukaryotic environment that supports proper folding, disulfide bond formation, and potential post-translational modifications. The dual N-terminal 10xHis-tag and C-terminal Myc-tag are relatively small compared to the full-length protein (511 aa, ∼57 kDa) and may cause minimal steric interference. While the full-length protein contains all functional domains, the probability of correct folding with functional glucosyltransferase activity requires experimental validation of enzymatic activity.
1. In Vitro Enzyme Activity Characterization
This application carries a significant risk without functional validation. Glucosyltransferase activity requires precise active site formation and proper cofactor binding. If correctly folded and active (verified through enzymatic assays), the protein is suitable for activity characterization. If misfolded/inactive (unverified), activity measurements will yield biologically meaningless results.
2. Substrate Specificity and Kinetic Studies
Meaningful kinetic studies require native enzyme conformation and functional activity. If correctly folded and active (verified), the protein enables valid Km and Vmax determinations. If misfolded/inactive (unverified), kinetic parameters will be biologically irrelevant and misleading.
3. Antibody Development and Immunoassay Applications
This application is highly suitable as antibody development relies on antigenic sequence recognition rather than functional enzymatic activity. The full-length protein provides comprehensive epitope coverage for generating specific antibodies. The high purity (>85%) ensures minimal contamination issues.
4. Protein-Protein Interaction Studies
This application requires proper folding validation. Enzyme interactions with regulatory partners require native conformation. If correctly folded (verified), the protein may identify physiological interactions; if misfolded/unverified, there is a risk of non-specific binding or tag-mediated artefacts.
5. Comparative Enzyme Studies Across Citrus Species
Valid comparative studies require native enzyme conformation and functional activity. If correctly folded and active (verified), the protein enables meaningful evolutionary comparisons; if misfolded/inactive (unverified), comparative analyses would yield misleading insights.
Final Recommendation & Action Plan
The baculovirus-expressed glucosyltransferase with small dual tags has a high probability of correct folding due to the eukaryotic expression system, but experimental validation of enzymatic activity is crucial. Begin with functional validation using standard glucosyltransferase assays (e.g., with limonoid substrates and UDP-sugar donors) to confirm specific activity before considering functional applications. Applications 1, 2, 4, and 5 require this validation. Application 3 (antibody development) can proceed immediately. The baculovirus system is particularly suitable for this plant enzyme as it provides proper eukaryotic folding environment. For reliable glucosyltransferase research, validate kinetic parameters and substrate specificity before proceeding with comparative or interaction studies.
