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Kinetics of spheroplasts formation from selected yeast strains
Zentralblatt Fur Mikrobiologie
|January 1, 1987
Summary
Yeast spheroplast formation using zymolyase enzyme was studied. Reaction rates varied significantly between yeast strains, with K. fragilis showing 80-fold higher susceptibility than S. cerevisiae.
Area of Science:
- Microbiology
- Biochemistry
- Enzymology
Background:
- Spheroplasts are essential for yeast genetic manipulation.
- Understanding yeast cell wall lysis kinetics is crucial for optimizing protoplast formation.
Purpose of the Study:
- To investigate the kinetics of spheroplast formation in Kluyveromyces and Saccharomyces yeasts.
- To compare the susceptibility of different yeast strains to zymolyase.
Main Methods:
- Yeast strains (Kluyveromyces and Saccharomyces) were treated with zymolyase.
- Cell lysis was monitored using the Coulter counter technique.
- Reaction kinetics were analyzed, comparing with spectrophotometric methods.
Main Results:
- Spheroplast formation followed first-order kinetics after an initial lag phase.
- Lysis rates were directly proportional to zymolyase concentration.
- Kluyveromyces fragilis exhibited 80-fold higher susceptibility to zymolyase compared to Saccharomyces cerevisiae CBS 5495.
- Spectrophotometric methods underestimated the reaction course compared to cell sizing.
Conclusions:
- Significant variations in yeast strain susceptibility to zymolyase exist.
- The Coulter counter technique provides accurate monitoring of yeast spheroplast formation.
- Enzyme concentration is a key factor in controlling yeast cell lysis rates.