Abstract
Bacterial peptidoglycan is the cell wall component responsible for maintaining cell integrity against osmotic pressure. Biosynthesis of the cytoplasmic precursor UDP-N-acetylmuramyl pentapeptide is catalyzed by the Mur enzymes. Genomic analysis of the three regions encoding Mur proteins was achieved. We have cloned and over-expressed the murA, -B, -D, -E and -F genes of Pseudomonas aeruginosa in pET expression system by adding a His-Tag to the C-termini of the proteins. Mur proteins were purified to homogeneity by a single chromatographic step on affinity nickel columns. Protein identities were verified through N-terminal sequencing. Enzyme activity was proved by the identification of the pathway's final product.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism
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Cell Wall / metabolism
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Chromatography, Affinity
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Cloning, Molecular
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Genes, Bacterial / genetics*
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Molecular Sequence Data
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Peptidoglycan / biosynthesis*
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Peptidoglycan / genetics
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Pseudomonas aeruginosa / enzymology
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Pseudomonas aeruginosa / genetics*
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Pseudomonas aeruginosa / growth & development
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Pseudomonas aeruginosa / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sequence Analysis, DNA
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Sequence Analysis, Protein
Substances
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Bacterial Proteins
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Peptidoglycan
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Recombinant Fusion Proteins