Research of Efficient Preparation of Laver Protoplasts by Compound Enzymatic Method

Laver (Porphyra yezoensis) was chosen for enzymatic hydrolysis, and protoplasts were obtained by enzyme hydrolysis. Effects on the enzyme of temperature, the amount of enzyme, reaction time, rotary speed, salinity, the proportion of components in the enzyme and metal ion were studied by the number of laver protoplasts. Factors and levels were selected on the basis of single factor test, the optimization of hydrolysis conditions were obtained by orthogonal experiment with the number of protoplasts as the index. The results indicated that the optimal hydrolytic conditions were:Temperature 28℃, rotary speed 125 r/min, content of cellulose crude enzyme 3%, the agar enzyme 12 mL, salinity 4% and hydrolysis time 100 min. Protoplast formation inhibited by some metal ions were found simultaneously. Protoplasts obtained under the optimal hydrolysis conditions were high live rate, the number of protoplasts obtained from 1-g porphyra sample was (16.88±0.22)×10⁶ cells/g.