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Lytic enzyme produced by Pseudomonas aeruginosa concomitantly with bacteriophage PS17. Purification,
Abstract:
A bacteriolytic enzyme was found to be produced, concomitantly with the progeny phage, in Pseudomonas aeruginosa P14 infected with phage PS17. The enzyme, named PS17-lysozyme, was purified by acrinol treatment, two cycles of Amberlite CG-50 chromatography, and SP-Sephadex C-50 chromatography. Homogeneity of the preparation was demonstrated by three electrophoretic techniques. PS17-lysozyme behaved like a basic protein (pI, 9-10) consisting of a single polypeptide chain (molecular weight, 24,500) and showed the substrate specificity as hen egg-white lysozyme. The enzyme exhibited much higher specific activity than the egg-white enzyme when assayed with chloroform-killed P. aeruginosa P14 as a substrate. These characteristics, as well as the amino acid composition, were very similar to those of PR1-lysozyme; a bacteriolytic enzyme produced in mitomycin C-induced P. aeruginosa P15 concomitantly with a phage-tail-like bacteriocin, pyocin R1 (Ochi et al. (1978) J. Biochem. 83, 727-736). However, the behavior of these two lysozymes from P. aeruginosa in Amberlite CG-50 chromatography and some other properties indicated that they were not identical, though they were similar. The results are in accord with the view that pyocin R1 may be a defective form of a bacteriophage closely related to but not identical with phage PS17.
Insights
A novel bacteriolytic enzyme, PS17-lysozyme, was purified from Pseudomonas aeruginosa infected with phage PS17. This enzyme shows high specific activity and shares similarities with PR1-lysozyme, suggesting a potential relationship between pyocin R1 and phage PS17.
Area of Science:
- Microbiology
- Enzymology
- Bacteriology
Background:
- Pseudomonas aeruginosa infections can be treated with bacteriophages and their associated enzymes.
- Bacteriolytic enzymes play a crucial role in phage replication and bacterial cell lysis.
Purpose of the Study:
- To purify and characterize a bacteriolytic enzyme produced during Pseudomonas aeruginosa infection with phage PS17.
- To compare the properties of this enzyme with other known lysozymes, particularly PR1-lysozyme.
Main Methods:
- Purification of PS17-lysozyme using acrinol treatment and multiple chromatography steps (Amberlite CG-50, SP-Sephadex C-50).
- Homogeneity assessment via three electrophoretic techniques.
- Characterization of enzyme properties including pI, molecular weight, substrate specificity, and amino acid composition.
Main Results:
- PS17-lysozyme was purified to homogeneity, revealing it to be a basic protein (pI 9-10) with a molecular weight of 24,500 Da.
- The enzyme demonstrated substrate specificity similar to hen egg-white lysozyme but exhibited significantly higher specific activity against P. aeruginosa P14.
- While similar to PR1-lysozyme, distinct chromatographic behavior and other properties indicated they are not identical.
Conclusions:
- PS17-lysozyme is a distinct bacteriolytic enzyme produced by phage PS17-infected Pseudomonas aeruginosa.
- The findings support the hypothesis that pyocin R1 might be a defective bacteriophage related to, but not identical with, phage PS17.