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Enterococcus faecalis V583 contains a cytochrome bd-type respiratory oxidase
L Winstedt1, L Frankenberg, L Hederstedt
1Department of Microbiology, Lund University, Lund, Sweden. lena.winstedt@mikrbiol.lu.se
Journal of Bacteriology
|June 13, 2000
Summary
Researchers cloned the Enterococcus faecalis cydABCD gene cluster, enabling aerobic respiration in Bacillus subtilis via a functional cytochrome bd terminal oxidase. This demonstrates E. faecalis
Area of Science:
- Microbiology and Molecular Biology
- Bacterial Respiration
- Gene Expression
Background:
- Enterococcus faecalis is a facultative anaerobe.
- Cytochrome bd oxidases are crucial for bacterial respiration.
- Heme is essential for the function of many respiratory enzymes.
Purpose of the Study:
- To clone and characterize the Enterococcus faecalis cydABCD gene cluster.
- To investigate the functional expression of the E. faecalis cytochrome bd oxidase in a heterologous host.
- To determine the role of heme in E. faecalis aerobic respiration.
Main Methods:
- Gene cloning and manipulation.
- Heterologous expression in Bacillus subtilis.
- Assessment of cytochrome bd oxidase activity.
- Growth studies in the presence of heme.
Main Results:
- The Enterococcus faecalis cydABCD gene cluster was successfully cloned.
- Expression of cydABCD in Bacillus subtilis resulted in a functional cytochrome bd terminal oxidase.
- E. faecalis V583 cells demonstrated aerobic respiration capacity when supplied with heme.
Conclusions:
- The cydABCD gene cluster encodes a functional cytochrome bd terminal oxidase in Enterococcus faecalis.
- Heme is required for the aerobic respiratory capacity of E. faecalis V583.
- Heterologous expression is a viable method for studying bacterial respiratory enzymes.