Abstract: The purpose of the present study was to develop a method to disrupt lotus bee pollen cell walls by one-step multienzymatic hydrolysis. Preliminary experiments were designed to investigate the effect of hydrolysis with different dosages of cellulase alone, 0.8% celluase and different dosages of pectinase, 0.8% celluase, 0.4% pectinase and different dosages of xylanase, or 0.8% celluase, 0.4% pectinase, 0.2% xylanase and different dosages of papain on the disruption of lotus bee pollen cell walls. We found that a mixture of cellulase, pectinase, xylanase and papain at a ratio of 4:2:1:3 was the best enzyme for the disruption of lotus bee pollen cell walls. Further, four hydrolysis parameters including pH, total enzyme dosage, temperature and material-to-water ratio for disrupting lotus bee pollen cell walls with this mixture of enzymes were optimized employing quadratic orthogonal rotation combination design. A mathematical model describing cell disruption percentage as a function of these four parameters was established. The optimal enzymatic hydrolysis parameters were found as pH 4.0, 12‰ of total enzyme dosage, 45 ℃ of hydrolysis temperature, and 1:14 (g/mL) of material-to-water ratio, resulting in a cell disruption percentage of 89.21%.

Key words: lotus bee pollen, enzymes, cell wall disruption