Recombinant Salmonella heidelberg Monofunctional biosynthetic peptidoglycan transglycosylase (mtgA)
Description
Introduction to Peptidoglycan Transglycosylases
Peptidoglycan transglycosylases are enzymes crucial for the synthesis of peptidoglycan, a key component of bacterial cell walls. These enzymes catalyze the formation of glycosidic bonds between the sugar moieties of peptidoglycan, which is essential for maintaining bacterial cell shape and integrity. Among these enzymes, monofunctional biosynthetic peptidoglycan transglycosylases are specialized in synthesizing new peptidoglycan strands without the ability to hydrolyze existing ones.
Understanding Monofunctional Biosynthetic Peptidoglycan Transglycosylase (mtgA)
Monofunctional biosynthetic peptidoglycan transglycosylases, such as the mtgA enzyme, are involved in the polymerization of peptidoglycan precursors into the bacterial cell wall. These enzymes are distinct from bifunctional enzymes, which can both synthesize and hydrolyze peptidoglycan. The mtgA enzyme in Salmonella species, including Salmonella heidelberg, plays a critical role in cell wall synthesis, ensuring the structural integrity necessary for bacterial survival and proliferation.
Recombinant Salmonella heidelberg Monofunctional Biosynthetic Peptidoglycan Transglycosylase (mtgA)
Recombinant Salmonella heidelberg monofunctional biosynthetic peptidoglycan transglycosylase (mtgA) refers to a genetically engineered version of the mtgA enzyme produced in a laboratory setting. This recombinant enzyme is used for research purposes, such as studying peptidoglycan synthesis mechanisms, understanding bacterial cell wall dynamics, and exploring potential targets for antimicrobial therapies.
3.1. Function and Importance
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Cell Wall Synthesis: The mtgA enzyme is crucial for synthesizing new peptidoglycan strands, which are essential for maintaining the structural integrity of the bacterial cell wall.
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Bacterial Survival: By ensuring proper cell wall formation, mtgA contributes to bacterial survival and resistance against environmental stresses.
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Antimicrobial Targets: Understanding the function of mtgA can help in developing targeted antimicrobial therapies that disrupt bacterial cell wall synthesis.
Research Findings and Data
While specific data on recombinant Salmonella heidelberg mtgA might be limited, research on similar enzymes in other bacteria provides insights into their function and importance. For instance, studies on peptidoglycan synthesis highlight the role of transglycosylases in bacterial cell wall formation and maintenance.
4.1. Comparison of Peptidoglycan Synthesis Enzymes
| Enzyme Type | Function | Example Enzymes |
|---|---|---|
| Monofunctional Biosynthetic Transglycosylases | Synthesize new peptidoglycan strands | mtgA in Salmonella |
| Bifunctional Transglycosylases/Hydrolases | Synthesize and hydrolyze peptidoglycan | PBP2 in Staphylococcus aureus |
| Lytic Transglycosylases | Hydrolyze peptidoglycan for cell wall remodeling | LTGs in Vibrio cholerae |
References
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Monofunctional Biosynthetic Peptidoglycan Transglycosylases: These enzymes are specialized in synthesizing new peptidoglycan strands without hydrolyzing existing ones. While specific references to mtgA in Salmonella heidelberg are not readily available, general information on monofunctional transglycosylases can be found in scientific literature .
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Peptidoglycan Synthesis and Cell Wall Dynamics: Research on peptidoglycan synthesis and cell wall remodeling provides a broader context for understanding the role of enzymes like mtgA .
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Recombinant Proteins in Research: Recombinant proteins are widely used in research to study enzyme functions and develop therapeutic applications .
Product Specs
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Target Background
KEGG: seh:SeHA_C3623
Q&A
Here’s a structured collection of FAQs tailored for academic researchers investigating recombinant Salmonella Heidelberg monofunctional biosynthetic peptidoglycan transglycosylase (MtgA):
Advanced Research Questions
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How do conflicting reports on MtgA’s essentiality in Salmonella impact experimental design?
While MtgA is non-essential in E. coli under standard conditions , Salmonella studies suggest context-dependent essentiality during host infection . Researchers should:
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Conduct conditional knockdowns (e.g., CRISPRi) in intracellular Salmonella models.
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Compare PG composition (via LC-MS) between wild-type and ΔmtgA strains under stress (e.g., β-lactam exposure) .
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What methodological challenges arise when analyzing MtgA’s activity in vivo?
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PG Dynamics: Use fluorescent D-amino acid (FDAAs) probes to visualize PG synthesis in real time .
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Crosslink Analysis: Employ muramidase digestion followed by UPLC-MS to quantify glycan chain length and crosslinking (Table 1).
Table 1: Comparative PG features in Salmonella Heidelberg strains
| Strain | Avg. Glycan Chain Length | % L,D-Crosslinks | MtgA Activity (nmol/min/mg) |
|---|---|---|---|
| Wild-type | 25–30 disaccharides | 15% | 12.4 ± 1.2 |
| ΔmtgA | 15–20 disaccharides | 8% | Undetectable |
| Complemented | 24–28 disaccharides | 14% | 10.8 ± 0.9 |
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How do MtgA inhibitors differ from β-lactams in targeting PG biosynthesis?
MtgA inhibitors (e.g., moenomycin analogs) block glycan chain polymerization without affecting transpeptidation. Key considerations:
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Resistance Mechanisms: Salmonella may upregulate alternative flippases (e.g., RodA) or hydrolases (e.g., SagA) .
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Synergy Testing: Combine MtgA inhibitors with β-lactams to exploit cell wall vulnerability .
Data Contradiction Analysis
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Why do some studies report MtgA redundancy while others highlight its indispensability?
Discrepancies arise from:
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Species-Specific Roles: Salmonella may rely more on monofunctional enzymes during host adaptation compared to E. coli .
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Growth Conditions: MtgA becomes critical under osmotic stress or antibiotic exposure .
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Compensatory Pathways: Overexpression of SEDS-family proteins (e.g., RodA) can rescue ΔmtgA phenotypes .
Methodological Recommendations
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How to resolve low yields of active recombinant MtgA?
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Membrane Mimetics: Use nanodiscs or amphipols to stabilize MtgA during purification .
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Activity Assays: Optimize with synthetic lipid II analogs (e.g., C35-Lipid II) to bypass in vitro synthesis hurdles .
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What controls are critical for interpreting MtgA knockout phenotypes?
