Recombinant Lactococcus lactis subsp. cremoris Energy-coupling factor transporter transmembrane protein EcfT (ecfT)
Description
Role in ECF Transporter Function
ECF transporters mediate ATP-dependent uptake of vitamins and micronutrients in bacteria. EcfT serves as the transmembrane scaffold, coordinating energy transfer between ATP hydrolysis (via EcfA/A') and substrate binding (via S-components) .
Key Functional Attributes:
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Quaternary Structure: Forms a stable complex with EcfA, EcfA', and variable S-components (e.g., BioY for biotin, RibU for riboflavin) .
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Substrate Diversity: A single EcfT module can pair with multiple S-components, enabling transport of diverse substrates :
| S-Component | Substrate | Gene ID (L. lactis) |
|---|---|---|
| BioY | Biotin | llmg_1964 |
| RibU | Riboflavin | llmg_1195 |
| ThiT | Thiamine | llmg_0334 |
| PanT | Pantothenate | llmg_0542 |
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Mechanism: ATP hydrolysis by EcfA/A' induces conformational changes in EcfT, driving substrate translocation through the S-component .
Recombinant Production and Characterization
The recombinant EcfT protein (Product ID: RFL16651LF) is produced in E. coli with the following specifications :
| Parameter | Details |
|---|---|
| Form | Lyophilized powder |
| Storage Buffer | Tris/PBS-based buffer, 6% trehalose (pH 8.0) |
| Reconstitution | 0.1–1.0 mg/mL in sterile water, with 5–50% glycerol for long-term storage |
| Stability | Stable at -20°C/-80°C; avoid repeated freeze-thaw cycles |
Notable Features:
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Full-length sequence (1–266 aa) preserves native structural integrity.
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High purity (>90%) ensures reliability in biochemical assays .
4.1. Mechanistic Studies
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Structural Biology: EcfT’s interaction with S-components has been resolved via cryo-EM and X-ray crystallography, revealing dynamic substrate-binding interfaces .
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Transport Assays: Reconstitution of EcfT-containing complexes in proteoliposomes demonstrates ATP-dependent vitamin uptake .
4.2. Antimicrobial Targeting
ECF transporters are absent in humans, making EcfT a candidate for narrow-spectrum antibiotics. Inhibitors targeting the EcfT-S-component interface block vitamin uptake in pathogens like Lactobacillus delbrueckii .
4.3. Industrial Relevance
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Strain Engineering: Adaptive evolution of L. lactis under stress (e.g., cell envelope stressors) has identified mutations in EcfT-associated pathways, informing starter culture optimization .
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Probiotic Development: While not directly linked to EcfT, L. lactis subsp. cremoris strains modulate host immunity via TLR2/MyD88 signaling, highlighting the genus’s biomedical potential .
Future Directions
Product Specs
Note: We prioritize shipping the format currently in stock. However, if you have specific requirements for the format, please indicate them during order placement, and we will accommodate your request.
Note: All of our proteins are shipped with standard blue ice packs by default. If you require dry ice shipping, please inform us in advance, as additional fees will apply.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Target Background
KEGG: llm:llmg_0289
STRING: 416870.llmg_0289
