With the development of biological enzyme technology, enzymatic separation is widely used to obtain protoplasts. Commonly used enzyme preparations include pectinase, cellulase, xylanase and so on. Usually, the wall-breaking effect of compound enzyme is better than that of single enzyme.

Pectinase is an efficient biocatalyst, which is metabolized by Aspergillus Niger and produced by submerged fermentation. Pectinase is not a single enzyme, but contains three components: pectinesterase, polygalacturonase and pectin lyase. Under the synergistic effect of these enzymes, pectin, arabinan, hemicellulose, starch, protein and other insoluble substances in fruit juice can be decomposed to produce galacturonic acid, galactooligosaccharic acid, unsaturated galacturonic acid and galactooligosaccharic acid.

Cellulase: it is also an enzyme preparation extracted by microbial fermentation. The structure and function of cellulase from different sources are very different. Cellulase also contains hemicellulase, 4- glucanase, xylanase, cellobiase and other components in practical application. It plays a biocatalytic role in decomposing cellulose, which can hydrolyze cellulose into monosaccharides and polysaccharides. When applied to fruit and vegetable juice and fruit wine, it can destroy the cell wall and improve the juice yield.