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Schizosaccharomyces pombe Isolation Protocol

Ángel Benito1, Fernando Calderón1, Santiago Benito2

  • 1Department of Chemistry and Food Technology, Polytechnic University of Madrid, Madrid, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|February 10, 2018
PubMed

Insights

This study presents a new selective medium for isolating Schizosaccharomyces pombe yeast. The method leverages unique traits like resistance to inhibitors and malo-ethanolic fermentation for easier detection.

Area of Science:

  • Microbiology
  • Food Science
  • Industrial Biotechnology

Background:

  • Schizosaccharomyces pombe is a yeast species with industrial relevance but is difficult to isolate due to its low natural abundance.
  • Accurate isolation methods are crucial for identifying S. pombe, especially when it acts as a spoilage microorganism.
  • Existing isolation techniques face challenges due to the low incidence of S. pombe compared to other microbes.

Purpose of the Study:

  • To describe a novel selective-differential medium for the efficient isolation and selection of Schizosaccharomyces pombe strains.
  • To facilitate the detection of S. pombe, particularly in contexts where it is considered a spoilage organism.
  • To enable the isolation of S. pombe for various industrial applications.

Main Methods:

  • Development of a selective-differential medium incorporating agents targeting S. pombe's unique physiological characteristics.
  • Utilizing S. pombe's resistance to actidione and benzoic acid as selective agents.
  • Employing malic acid as a differential agent, detecting its exclusive metabolism into ethanol by S. pombe.
  • Incorporating cloramphenicol to inhibit bacterial growth and using liquid media to prevent fungal contamination.

Main Results:

  • The proposed medium effectively isolates S. pombe by exploiting its resistance to common inhibitory substances like actidione and benzoic acid.
  • Malic acid degradation serves as a reliable differential marker, distinguishing S. pombe from other microorganisms.
  • The methodology allows for the detection and isolation of S. pombe even at low natural incidences.
  • Bacterial contamination is minimized through the inclusion of cloramphenicol.

Conclusions:

  • The developed selective-differential medium significantly enhances the isolation and selection of Schizosaccharomyces pombe.
  • This method provides a robust tool for identifying S. pombe in industrial settings and as a spoilage agent.
  • The approach leverages specific physiological traits of S. pombe for improved detection accuracy and efficiency.
Keywords:
ActidioneBenzoic acidCloramphenicolDifferential selective mediumMalic acidSchizosaccharomyces pombe

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