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

Cellulose Filtration Agar Asbestos 

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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