A Lallzyme MMX-based rapid method for fission yeast protoplast preparation

Area of Science:

• Microbiology
• Cell Biology

Background:

• Yeast cells possess a cell wall essential for structural integrity, counteracting turgor pressure.
• Cell wall removal is critical for various experimental procedures, including genetic manipulation and nucleic acid isolation.
• Generating live, cell wall-deficient protoplasts is vital for specific yeast research applications.

Purpose of the Study:

• To develop a rapid and efficient method for preparing fission yeast protoplasts.
• To overcome limitations of existing time-consuming and inefficient cell wall digestion protocols.
• To provide a robust protoplast preparation technique applicable to common fission yeast experiments.

Main Methods:

• Utilized Lallzyme MMX, a commercial enzyme mix, for rapid enzymatic digestion of fission yeast cell walls.
• Developed a protocol achieving protoplast generation in less than 10 minutes.
• Validated the utility of the prepared protoplasts in downstream applications.

Main Results:

• Achieved efficient protoplast preparation from all fission yeast cells within 10 minutes.
• Demonstrated successful application of protoplasts in plasmid extraction, diploid generation, and morphogenesis assays.
• Established a new, robust, and rapid protocol for fission yeast protoplast generation.

Conclusions:

• The new Lallzyme MMX-based method offers a significant improvement in speed and efficiency for fission yeast protoplast preparation.
• The generated protoplasts are viable and suitable for critical downstream applications in fission yeast research.
• This method has the potential for broader application in other yeast and fungal species.