Recombinant Daucus carota Peroxidase 10
Description
Molecular and Genetic Characteristics
Recombinant Daucus carota Peroxidase 10 is a 17.5 kDa protein (calculated molecular weight) encoded by the LOC108193863 gene, annotated as a peroxidase 10-like protein in the carrot genome . Key molecular properties include:
| Parameter | Details | Source |
|---|---|---|
| Molecular Weight | 1,746 Da (recombinant form) | |
| Host Systems | E. coli, Yeast, Baculovirus, Mammalian Cells | |
| Purity | ≥85% (SDS-PAGE verified) | |
| Gene ID | LOC108193863 (peroxidase 10-like) |
The gene resides within the carrot genome, which has undergone telomere-to-telomere sequencing to resolve structural gaps, enabling precise annotation of peroxidase-related loci .
Production and Purification
The recombinant protein is synthesized via heterologous expression systems, commonly E. coli, and purified using chromatography techniques:
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Expression: Host systems vary, with E. coli being the most frequently cited .
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Purification: Sequential ion-exchange and affinity chromatography (e.g., Ni-affinity for His-tagged variants) .
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Yield: Purity exceeds 85% as confirmed by Coomassie-stained SDS-PAGE .
This production strategy mirrors methods used for other plant-derived recombinant proteins, such as miraculin in carrot callus cultures .
Biological Functions
Peroxidase 10 performs diverse roles in plant physiology, as inferred from UniProt annotations and functional studies:
| Function | Mechanism | Relevance |
|---|---|---|
| H₂O₂ Removal | Catalyzes oxidation of hydrogen peroxide | Mitigates oxidative stress |
| Lignin Biosynthesis | Oxidizes phenolic precursors (e.g., coniferyl alcohol) | Structural reinforcement in cell walls |
| Stress Response | Activated by oxidative stress (e.g., H₂O₂, NaCl) | Adaptive defense against abiotic stress |
| Auxin Catabolism | Regulates auxin levels via oxidation | Modulates growth and development |
While substrate specificity for Recombinant Daucus carota Peroxidase 10 is not explicitly detailed, peroxidases generally oxidize phenolic compounds, as demonstrated in Asparagus peroxidase studies .
Research Applications
This enzyme is employed in experimental settings to study:
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Oxidative Stress Pathways: Utilized to model ROS detoxification in plant cells .
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Lignin Engineering: Investigated for its role in modifying lignin content, critical for biofuel production .
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Protein Expression Systems: Serves as a model for optimizing recombinant protein production in carrot hosts .
Example Workflow:
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ELISA/Western Blot: Detection of Peroxidase 10 in plant extracts or recombinant systems .
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Enzymatic Assays: Evaluation of substrate oxidation kinetics (e.g., guaiacol, coniferyl alcohol) .
Genomic Context
The carrot genome assembly (D. carota vT2T) has resolved structural gaps, enabling precise mapping of peroxidase genes . Key genomic features include:
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Repeat Content: 55.30% of the genome, dominated by LTR transposons .
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Gene Annotation: 96.40% of predicted genes match public databases (e.g., UniProt) .
This genomic resolution aids in identifying orthologs and regulatory elements, such as the SWPA2 promoter used in stress-inducible expression systems .
