Recombinant Pelotomaculum thermopropionicum UPF0059 membrane protein PTH_2827 (PTH_2827)
Description
Genomic Context and Functional Annotation
Pelotomaculum thermopropionicum is a Gram-positive thermophile specializing in syntrophic propionate oxidation under anaerobic conditions . Its genome (3,025,375 bp) contains 2,920 coding sequences (CDSs), with PTH_2827 annotated as a hypothetical membrane protein belonging to the UPF0059 family .
Key Genomic Features of PTH_2827:
The gene is located on the leading strand of the P. thermopropionicum genome, which is characteristic of Firmicutes bacteria . Despite its hypothetical classification, its genetic proximity to sulfur metabolism-related clusters suggests a potential role in sulfur anion transport or membrane stabilization .
Recombinant Expression and Purification
Recombinant PTH_2827 is produced in E. coli using a T7 promoter system, with yields optimized through tunable expression strains like Lemo21(DE3) . Key challenges include:
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Hydrophobicity: Transmembrane domains (e.g., residues 1–180) necessitate detergent stabilization during purification .
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Expression Optimization: Low expression levels are mitigated using glycerol storage (50%) and temperature-controlled cultivation .
Example Purification Protocol:
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Cultivation: E. coli Lemo21(DE3) cells grown at 37°C in LB medium with 0.1–1.0 mM rhamnose for T7 RNA polymerase induction .
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Membrane Isolation: Cells lysed via sonication; membranes solubilized with n-dodecyl-β-D-maltoside (DDM) .
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Affinity Chromatography: His-tag purification under denaturing conditions (8 M urea) .
Functional Hypotheses:
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Electron Transport: Potential interaction with hydrogenases (e.g., PTH_0684–0686) .
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Sulfur Metabolism: Genetic proximity to sulfate assimilation clusters (e.g., cysH, cysN) .
Applications:
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Structural Biology: Crystallization trials for resolving membrane-protein interactions .
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Syntrophic Metabolism Studies: Role in interspecies electron transfer with methanogens .
Challenges:
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Low Solubility: Requires lipid nanodiscs or detergent micelles for stabilization .
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Functional Validation: No enzymatic activity reported to date; in vitro assays needed .
Future Directions
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Cryo-EM Studies: Resolve 3D structure to identify catalytic or binding sites.
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Heterologous Co-Expression: Co-culture with methanogens to validate syntrophic interactions .
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Biotechnological Applications: Engineer PTH_2827 for bioenergy applications (e.g., microbial fuel cells) .
References
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Genome analysis of P. thermopropionicum (PMC2259108).
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Membrane protein expression challenges (PMID:16777403).
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Genomic annotation of PTH_2827 (biocomputo.ibt.unam.mx).
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Recombinant PTH_2827 product details (proteinfoldingcenter.org).
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Difficult-to-express proteins (futurefields.io).
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Lemo21(DE3) optimization (NEB).
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Physiological characterization of P. thermopropionicum (PMID:12361280).
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E. coli expression systems (creative-biolabs.com).
Product Specs
Note: We will prioritize shipping the format that we have in stock. However, if you have specific requirements for the format, please indicate them in your order notes. We will fulfill your request if possible.
Note: Our proteins are shipped with standard blue ice packs. If you require dry ice shipping, please inform us in advance, as additional fees will apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Target Background
KEGG: pth:PTH_2827
STRING: 370438.PTH_2827
