Recombinant Enolase (eno)
Description
Definition and Biochemical Properties
Enolase is a metalloenzyme requiring two magnesium ions for catalytic activity. It exists as three isoforms in humans: ENO1 (α-enolase), ENO2 (γ-enolase), and ENO3 (β-enolase). Recombinant ENO1, the most studied isoform, is a homodimer with a molecular weight of approximately 93 kDa when expressed in E. coli . Its sequence homology across species (40–90%) underscores its evolutionary conservation .
| Property | Details | Source |
|---|---|---|
| Isoforms | α, β, γ | |
| Substrate | 2-PGA → PEP | |
| Cofactors | Mg²⁺ ions | |
| Molecular Weight | ~93 kDa (dimer) |
Production Methods
Recombinant ENO is typically produced via bacterial fermentation. E. coli systems are favored due to their scalability and compatibility with affinity chromatography.
| Step | Details | Source |
|---|---|---|
| Vector | pGEMEX-2 or pET-28a | |
| Purification | Ni²⁺-affinity, SEC | |
| Purity | >90% (SDS-PAGE) | |
| Activity | 6,000 pmol/min/μg |
Vaccine Development
Recombinant ENO has shown efficacy as a vaccine antigen against tick-borne illnesses. Mice immunized with Haemaphysalis longicornis enolase exhibited 90% resistance to tick infestation .
| Target Pathogen | Efficacy | Source |
|---|---|---|
| H. longicornis | 90% reduction in tick attachment | |
| Sporothrix spp. | Induces humoral immunity |
Therapeutic Use
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Cancer: ENO1 is overexpressed in metastatic tumors, making it a target for inhibitors .
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Inflammation: Recombinant ENO reduced synovitis in arthritis models .
Diagnostics
ENO levels in cerebrospinal fluid correlate with astrocytoma severity and stroke prognosis .
Pathogen-Host Interactions
Ehrlichia chaffeensis degrades host ENO1 via ubiquitination to modulate glycolysis, a mechanism linked to infection persistence .
