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Water-soluble polysaccharide was isolated from Poria cocus by enzyme-assisted method using orthogonal methodology. By single factor test and orthogonal group, the extraction conditions of watersoluble polysaccharide were investigated, such as liquid-solid ratio, temperature, time and pH. The optimum conditions for single factor were as follows Enzyme concentration (%), Extraction temperature oC, Extraction time/h, Extraction pH. The result revealed that 2% complex enzyme, remained most important factor of polysaccharide extraction, followed by temperature and pH. The liquid-solid ratio was 1:50, temperature was 40°C, time was 3.0 h, and pH was 5. The highest extraction rate of crude polysaccharide remained 4.14%. Results indicated that the complex-enzyme assisted remained best technique for extracting polysaccharide from P. cocus. It proved to be as highly effective as well as energy and time saving extraction techniques.

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