Recombinant Stenotrophomonas maltophilia Glycine dehydrogenase [decarboxylating] (gcvP), partial
Description
Introduction to Glycine Dehydrogenase (gcvP)
Glycine dehydrogenase [decarboxylating] (gcvP) is a mitochondrial or bacterial enzyme that catalyzes the decarboxylation of glycine in the glycine cleavage system (GCS). In Stenotrophomonas maltophilia, a Gram-negative opportunistic pathogen, gcvP plays a critical role in one-carbon metabolism, linking amino acid catabolism to folate-mediated pathways. Recombinant versions of this enzyme enable detailed biochemical studies and applications in metabolic engineering or antimicrobial research.
Functional Role of gcvP in S. maltophilia
The glycine cleavage system, including gcvP, is essential for:
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Amino acid degradation: Converts glycine into 5,10-methylenetetrahydrofolate, NADH, and CO .
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Metabolic flexibility: Supports adaptation to nutrient-limited environments, such as host tissues .
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Pathogenicity: Linked to bacterial survival under oxidative stress and iron-scavenging mechanisms .
Recombinant Production of gcvP
Recombinant gcvP is typically expressed in Escherichia coli systems due to their scalability and compatibility with bacterial enzymes . Key steps include:
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Cloning: The gcvP gene (partial or full-length) is inserted into plasmids with affinity tags (e.g., hexahistidine) for purification.
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Expression: Induced using IPTG in optimized media (e.g., LB broth) .
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Purification: Nickel-affinity chromatography isolates the enzyme .
Table 1: Production Parameters for Recombinant gcvP
| Parameter | Details |
|---|---|
| Host System | E. coli BL21(DE3) |
| Vector | pET-28a(+) with T7 promoter |
| Tag | N-terminal hexahistidine |
| Induction | 0.5 mM IPTG at OD = 0.6–0.8, 18–24 hrs at 18°C |
| Purity | >90% (SDS-PAGE verified) |
Table 2: Enzymatic Properties (Inferred from Homologs)
| Property | Value |
|---|---|
| Optimal pH | 7.5–8.5 |
| Temperature Stability | Up to 40°C |
| (Glycine) | ~0.2 mM |
| Inhibitors | Methotrexate, aminopterin |
Applications in Research
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Antimicrobial Target: gcvP is a potential target for inhibiting S. maltophilia metabolism, given its role in folate and iron acquisition .
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Biocatalysis: Used in synthetic pathways for glycine-derived compounds (e.g., serine, purines) .
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Diagnostics: Recombinant gcvP aids in developing enzyme-linked assays for bacterial detection .
Challenges and Future Directions
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Partial Sequence Limitations: Truncated recombinant gcvP may lack full activity or regulatory domains .
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Antibiotic Resistance Context: S. maltophilia’s intrinsic multidrug resistance complicates inhibitor design .
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Structural Studies: Cryo-EM or X-ray crystallography is needed to resolve mechanistic details .
Product Specs
Target Background
KEGG: sml:Smlt3579
STRING: 522373.Smlt3579
