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Beta-glucuronidases of clostridium perfringens
Zentralblatt Fur Bakteriologie, Mikrobiologie, Und Hygiene. Series A, Medical Microbiology, Infectious Diseases, Virology, Parasitology
|August 1, 1984
Summary
This study isolated and purified two forms of beta-glucuronidase from Clostridium perfringens. Characterization revealed distinct properties and Michaelis constants for each enzyme, offering insights into their biochemical behavior.
Area of Science:
- Microbiology
- Enzymology
- Biochemistry
Background:
- Clostridium perfringens is a bacterium found in the gut.
- Beta-glucuronidase is an enzyme with various biological roles.
Purpose of the Study:
- To isolate and purify extracellular beta-glucuronidase from Clostridium perfringens.
- To characterize the biochemical properties of the purified enzymes.
Main Methods:
- Culturing Clostridium perfringens in GAM broth.
- Enzyme purification using ammonium sulfate precipitation, DEAE cellulose chromatography, Sephadex G-200 gel filtration, and affinity chromatography.
- Characterization of enzyme properties including pH optima, stability, molecular weight, and Michaelis constants.
Main Results:
- Two distinct beta-glucuronidase enzymes (I and II) were purified.
- Beta-glucuronidase I: optimum pH 7.2, molecular weight 115,000.
- Beta-glucuronidase II: optimum pH 6.0, molecular weight 195,000.
- Both enzymes were inhibited by Cu++ and Hg++, with inhibition reversed by cysteine; EDTA had no effect.
- Michaelis constants were determined for three different substrates.
Conclusions:
- Clostridium perfringens produces at least two forms of beta-glucuronidase with differing biochemical characteristics.
- The purified enzymes exhibit distinct pH optima, molecular weights, and substrate affinities.
- Understanding these enzymes can contribute to the study of microbial metabolism and enzyme function.