M. Serrano et al., EFE ACTIVITY IN PROTOPLASTS .1. PREPARATION OF KIWI PROTOPLASTS AND MEASUREMENT OF ITS EFE ACTIVITY, Revista espanola de ciencia y tecnologia de alimentos, 33(3), 1993, pp. 299-306
The obtaining of kiwi protoplasts and the influence of factors affecti
ng it, such as pH, concentration of the hydrolytic enzymes caylase and
pectolyase, concentration of sorbitol and digestion time were studied
. The highest amount of protoplasts per gram of tissue were obtained b
y the following digestion medium: EPPS-Tris buffer, 100 mM (pH 8.40) c
ontaining caylase 0.10%, pectolyase 0.01%, Cl2Ca 10 mM and sorbitol 0.
70 M. The most appropriate digestion time was 2 h. The reproducibility
of the obtaining of protoplasts was tested. It was verified that the
protoplast were able to convert the 1-aminocyclopropane-1-carboxylic a
cid into ethylene. The EFE activity in the protoplasts was quite high
when they were obtained in the presence of I-aminocyclopropane-1-carbo
xylic acid (ACC) 1 mM, but it became almost completely lost when the d
igestion and washing were performed without ACC.