Recombinant Jannaschia sp. Monofunctional biosynthetic peptidoglycan transglycosylase (mtgA)
Description
Introduction to Recombinant Jannaschia sp. Monofunctional Biosynthetic Peptidoglycan Transglycosylase (mtgA)
Recombinant Jannaschia sp. Monofunctional Biosynthetic Peptidoglycan Transglycosylase (mtgA) is a recombinant protein expressed in Escherichia coli and is involved in the biosynthesis of peptidoglycan, a critical component of bacterial cell walls. This enzyme plays a crucial role in the elongation of glycan chains during peptidoglycan synthesis, which is essential for bacterial cell wall integrity and survival.
Structure and Function
The recombinant mtgA protein is a full-length monofunctional transglycosylase, meaning it only catalyzes the transglycosylation reaction without participating in transpeptidation. It is composed of 239 amino acids and is fused with an N-terminal His tag to facilitate purification and detection . The His tag allows for easy affinity chromatography, making it convenient for researchers to isolate and study the protein.
Expression and Purification
The mtgA protein is expressed in E. coli, which serves as a common host for recombinant protein production due to its well-characterized genetics and ease of cultivation. The use of E. coli allows for large-scale production of mtgA, enabling extensive biochemical and structural studies.
Biochemical Characteristics
While specific biochemical characteristics of mtgA, such as its kinetic parameters or substrate specificity, are not detailed in the available literature, monofunctional transglycosylases generally require metal ions for catalytic activity, similar to other enzymes involved in peptidoglycan synthesis . Further research would be necessary to fully elucidate these properties for mtgA.
Research Findings and Applications
Research on monofunctional transglycosylases like mtgA contributes to understanding bacterial cell wall synthesis and its potential as a target for antibiotic development. The study of these enzymes can provide insights into how bacteria maintain their cell walls and how this process can be disrupted.
Data Tables
| Characteristic | Description |
|---|---|
| Protein Length | 239 amino acids |
| Expression Host | Escherichia coli |
| Tag | N-terminal His tag |
| Function | Monofunctional biosynthetic peptidoglycan transglycosylase |
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized fulfillment.
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The tag type is finalized during production. If you require a specific tag, please inform us for preferential development.
Target Background
KEGG: jan:Jann_0276
STRING: 290400.Jann_0276
Q&A
Basic Research Questions
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What is Jannaschia sp. and what are its ecological characteristics?
Jannaschia sp. is a genus within the Roseobacter group of Alphaproteobacteria. Most characterized strains are marine bacteria isolated from seawater environments. For example, Jannaschia sp. strains AI_61 and AI_62 were isolated from seawater at a depth of 1 m around a coastal aquaculture in Ainan, Ehime, Japan (32°56 N, 132°30 E) . Another species, Jannaschia rubra, was isolated from Mediterranean sea water near Valencia, Spain .
Jannaschia species typically display aerobic, chemo-organotrophic metabolism, with some strains exhibiting characteristics of aerobic anoxygenic phototrophy. For instance, strains AI_61 and AI_62 produce bacteriochlorophyll a (BChl a), indicated by a specific absorbance peak at 770 nm when extracted using the acetone-methanol (70:30, vol/vol) method .
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What is the function of monofunctional biosynthetic peptidoglycan transglycosylase (mtgA) in bacterial cell wall synthesis?
Monofunctional biosynthetic peptidoglycan transglycosylase (mtgA) catalyzes a critical step in bacterial cell wall biosynthesis. The enzyme specifically performs the transglycosylase activity (EC 2.4.2.-) , which involves the polymerization of peptidoglycan glycan strands through the formation of β-1,4-glycosidic bonds between lipid II precursors.
Unlike bifunctional enzymes that possess both transglycosylase and transpeptidase activities, mtgA is specialized for the glycan strand polymerization step only. This specificity makes it valuable for researchers studying the discrete steps of peptidoglycan synthesis and as a potential target for antimicrobial development.
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What are the optimal storage and handling conditions for recombinant Jannaschia sp. mtgA?
According to product information , the recombinant mtgA protein should be handled as follows:
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Storage Buffer: Tris-based buffer with 50% glycerol, optimized for this protein
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Short-term Storage: Store working aliquots at 4°C for up to one week
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Long-term Storage: Store at -20°C; for extended storage, conserve at -20°C or -80°C
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Important Note: Repeated freezing and thawing is not recommended
These storage conditions are essential to maintain the structural integrity and enzymatic activity of the recombinant protein for experimental purposes.
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How does the genomic context of the mtgA gene in Jannaschia sp. compare to related bacteria?
The mtgA gene in Jannaschia sp. (strain CCS1) is designated with the ordered locus name Jann_0276 . While the specific genomic context is not detailed in the provided search results, other related marine bacteria in the Roseobacter clade have had their genomes extensively sequenced.
For instance, the genomes of Jannaschia sp. strains AI_61 and AI_62 have been sequenced, consisting of 39 and 49 scaffolds respectively, with total lengths of 3,963,206 bp and 3,918,448 bp . These genomes contain approximately 3,800 protein-coding sequences each, as identified by DFAST and Prokka analyses.
Comparative genomic analysis could reveal conserved gene neighborhoods and potential operonic structures that include mtgA, providing insights into its regulation and functional relationships.
