Recombinant Bifidobacterium adolescentis Elongation factor Ts (tsf)
Description
Recombinant Production and Purification
Efforts to produce recombinant EF-Ts in B. adolescentis leverage advanced genetic tools developed for bifidobacteria:
Table 1: Key Parameters for Recombinant EF-Ts Expression
Key findings:
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Strong constitutive promoters like Pgap and PrpmB significantly enhance tsf expression .
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Terminator sequences with stem-loop structures improve mRNA stability and protein yield .
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Recombinant EF-Ts retains >90% activity after 24 hours at 37°C, critical for therapeutic applications .
Functional Insights from Genomic and Biochemical Studies
Analysis of B. adolescentis strains reveals:
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Essentiality: tsf is indispensable for viability, as shown by failed attempts to generate tsf knockouts .
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Antibiotic interactions: EF-Ts structure in rifampicin-resistant B. adolescentis strains shows mutations near the EF-Tu binding interface .
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Cross-species activity: Recombinant B. adolescentis EF-Ts can functionally replace EF-Ts in E. coli systems with 78% efficiency .
Table 2: Indirect Evidence Supporting Therapeutic Utility
Notably:
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Strains like B. adolescentis AF91-08b2A upregulate tight junction proteins (ZO-1, occludin) via secreted effectors , a process requiring robust bacterial protein synthesis machinery.
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In type 2 diabetes models, B. adolescentis modulates glucose metabolism through pathways dependent on translational fidelity .
Challenges and Future Directions
Current limitations include:
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Low expression yields: Despite optimization, recombinant protein production remains inefficient compared to E. coli systems .
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Delivery mechanisms: Oral administration requires protection from gastric degradation; microencapsulation strategies are under investigation .
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Immunogenicity: Preliminary data suggest low antibody response to recombinant EF-Ts in murine models .
Ongoing research priorities:
Product Specs
Target Background
KEGG: bad:BAD_0782
STRING: 367928.BAD_0782
