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Mutanolysin-induced lysis of actinomyces pyogenes determined by aggregometry
Abstract:
The lytic activity of mutanolysin from Streptomyces globisporus on 42 cultures of Actinomyces pyogenes could be effectively analyzed in an aggregometer. It was expressed as increase of transmittance at 546 nm after 20 min and 2 h at 37 degrees C. The A. pyogenes cultures revealed no uniform lysis pattern. Most of the cultures were lyzed within 20 to 40 min at 37 degrees C, others were lyzed only moderately or weakly within 2 h of incubation. The lytic activity was optimal at low (0.01 mol/l) molarity of the lysis buffer between pH 5.7 and 7 and could be inhibited by HgCl2.A. pyogenes was not lyzed by lysostaphin or lysozyme.
Insights
Mutanolysin from Streptomyces globisporus effectively lyses Actinomyces pyogenes cultures, though lysis patterns vary. Optimal conditions for mutanolysin activity were identified, showing inhibition by HgCl2.
Area of Science:
- Microbiology
- Enzymology
- Biochemistry
Background:
- Actinomyces pyogenes is a significant pathogen.
- Understanding bacterial lysis is crucial for developing targeted therapies.
- Mutanolysin is an enzyme with known lytic properties.
Purpose of the Study:
- To analyze the lytic activity of mutanolysin on Actinomyces pyogenes.
- To determine optimal conditions for mutanolysin's lytic action.
- To compare mutanolysin's efficacy with other lytic enzymes.
Main Methods:
- Lytic activity was assessed using an aggregometer measuring changes in transmittance at 546 nm.
- Experiments were conducted at 37°C over 20 minutes and 2 hours.
- Lysis buffer molarity and pH were varied, and inhibition by HgCl2 was tested.
Main Results:
- Mutanolysin exhibited effective lysis of Actinomyces pyogenes cultures, with varying lysis patterns observed.
- Optimal lytic activity occurred at low buffer molarity (0.01 mol/l) and pH between 5.7 and 7.
- HgCl2 inhibited mutanolysin activity, while lysostaphin and lysozyme showed no lysis against A. pyogenes.
Conclusions:
- Mutanolysin demonstrates potent lytic activity against Actinomyces pyogenes.
- Specific buffer conditions optimize mutanolysin's efficacy.
- Mutanolysin's specificity is highlighted by its activity where other enzymes fail.