Enzymes for preparation of protoplasts

Protoplast isolation is achieved by using cellulase in combination with pectinase and hemicellulase.

Commercial preparations of enzymes are derived from microorganisms, and may contain ribonucleases, proteases, and several other toxic enzymes. As a result of these deleterious enzymes, several purification procedures have been developed (one of the most often used procedures is column separation).
There are two approaches to the use of wall-degrading enzymes.
  1. Mixed-enzyme method: pectinase and cellulase and/or hemicellulase are applied simultaneously.
  2. Sequential method: involves treatment with pectinase (loosen the cells), followed by cellulase and or hemicellulase.

Commercial Enzyme Preparations
Onozuka R10, cellulase
Cellulysin, cellulase
Driselase, cellulase
Macerozyme R10, pectinase
Pectinase
Rhozyme HP150, hemicellulase
PATE (pectic acid acetyl transferase)