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Enzymes for Digestion of Yeast Ascus Walls

  • John F. T. Spencer
  • Dorothy M. Spencer
  • I. J. Bruce

Abstract

Snail enzyme (glusulase, helicase), zymolyase, Novozyme, etc., are all satis­factory. These enzymes can also be used for removal of walls from vegetative yeast cells (see section on protoplasts and protoplast fusion, and on trans­formation). A suitable enzyme for digestion of ascus walls can be extracted from commercial mushrooms, as follows:
  1. 1.

    Cut up 450–500 g of commercial mushrooms and homogenize in 500 ml of distilled water.

     
  2. 2.

    Strain off solid residues through gauze.

     
  3. 3.

    Centrifuge the liquid at 10–20,000 g and collect the supernatant and allow to stand at 4 °C overnight.

     
  4. 4.

    Centrifuge again at high speed.

     
  5. 5.

    Collect supernatant and sterilize by filtration. Cellulose membrane fil­ters are not suitable, for obvious reasons. Use asbestos filter mats or similar material (Ford Sterimat or Seitz filters).

     
  6. 6.

    Store frozen, in 10–20-ml aliquots.

     
  7. 7.

    The crude enzyme can be purified by standard methods for protein purification.

     

Keywords

Protoplast Fusion Cellulose Membrane Needle Holder Meiotic Gene Yeast Genetic 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • John F. T. Spencer
    • 1
  • Dorothy M. Spencer
    • 1
  • I. J. Bruce
    • 2
  1. 1.Department of Life SciencesGoldsmith’s CollegeLondonGreat Britain
  2. 2.Department of Biology and Environmental HealthThames PolytechnicLondonGreat Britain

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