Part:BBa_K5243001
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Oxalate Oxidase Expression Plasmid Parts Registry
Overview
This plasmid is designed for the expression of **oxalate oxidase** in Escherichia coli. Oxalate oxidase, also known as Germin, is an enzyme that catalyzes the breakdown of oxalate into carbon dioxide and hydrogen peroxide, helping to mitigate conditions such as hyperoxaluria. The following registry outlines the major genetic components of the plasmid, providing detailed descriptions of each part’s role in the expression system.
| Part Code | Part Name | Type | Function |
|---|---|---|---|
| BBa_K206000 | araBAD Promoter | Promoter | The araBAD promoter is an inducible promoter activated by the presence of L-arabinose in the growth medium. It tightly regulates the expression of the oxalate oxidase gene, allowing for control of the enzyme's production. This inducible system prevents the metabolic burden on E. coli before induction and enhances protein expression once arabinose is present. |
| BBa_B0034 | RBS (Ribosome Binding Site) | RBS | This ribosome binding site (RBS) ensures efficient translation of the mRNA by recruiting the ribosomes to the start codon of the oxalate oxidase gene. The sequence is optimized to achieve high levels of translation and maximize protein expression. |
| BBa_K5243000 | Oxalate Oxidase (Germin) | CDS (Coding Sequence) | The coding sequence of oxalate oxidase, an enzyme that degrades oxalate into hydrogen peroxide and carbon dioxide. This process is essential for preventing oxalate-related disorders like hyperoxaluria. The gene has been codon-optimized for expression in E. coli, ensuring efficient production and functionality of the enzyme. |
| BBa_K1319001 | TEV Cleavage Site | Tag | A TEV protease cleavage site is incorporated for easy removal of the GST tag after protein purification. This feature allows for precise cleavage and recovery of the oxalate oxidase enzyme in its native form, enhancing its utility for biochemical assays or therapeutic applications. |
| BBa_K1319002 | GST Tag | Tag | The GST (Glutathione S-transferase) tag allows for affinity purification of the oxalate oxidase enzyme using a glutathione-based matrix. This tag simplifies the purification process and ensures high yield and purity of the protein. |
| BBa_B0015 | rrnB Terminator | Terminator | The rrnB terminator efficiently terminates transcription, ensuring that mRNA is properly processed and that no unintended transcriptional read-through occurs. This ensures high fidelity in the gene expression system. |
| BBa_B1006 | Ampicillin Resistance Gene | Antibiotic Resistance | The ampicillin resistance gene confers resistance to the antibiotic ampicillin, allowing for the selection of E. coli cells that have successfully incorporated the plasmid. This marker ensures the growth of only transformed cells on selective media. |
| BBa_J61100 | pBR322 Origin of Replication | Origin of Replication | The pBR322 origin of replication ensures the plasmid is replicated in E. coli and maintained during cell division. This origin allows for stable maintenance of the plasmid at an optimal copy number within the bacterial cells. |
| BBa_I13458 | araC | Regulatory Protein | The araC gene encodes the AraC protein, which regulates the arabinose operon. It is responsible for activating the araBAD promoter in the presence of arabinose, ensuring the system is tightly regulated and that oxalate oxidase is only expressed under the appropriate conditions. |
Oxalate Oxidase: Germin
Oxalate oxidase, commonly known as Germin, is an enzyme that catalyzes the degradation of oxalate into carbon dioxide and hydrogen peroxide. This enzyme plays a vital role in maintaining oxalate homeostasis in plants by breaking down oxalate, which can chelate essential minerals such as calcium. In synthetic biology, the gene for oxalate oxidase has been codon-optimized for expression in E. coli to enable efficient production for various industrial and biomedical applications.
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