Enzymes for the Isolation of Protoplasts

Abstract

Cocking (1960) first used enzymes to isolate higher plant protoplasts. He applied culture filtrates of a cellulolytic fungus Myrothecium verrucaria to release protoplasts from tomato root tips. Later, Takebe and co-workers succeeded in applying commercially available cell wall-degrading enzymes, Macerozyme and Cellulase Onozuka, to isolate large quantities of protoplasts from a wide variety of plant tissues (Takebe et al. 1968; Otsuki and Takebe 1969). Thereafter, many enzmye preparations have been produced commercially and used successfully for the isolation of plant protoplasts. These enzmyes are divided into three categories: cellulases, hemicellulases and pectinases by their catalytic actions (Table 1). Combined use of these enzymes and development of isolation procedures have made