Purification of squalene-2,3-epoxide cyclases from cell suspension cultures of Rabdosia japonica Hara
Microsomes prepared from cell suspension cultures of Rabdosia japonica Hara showed activities for cyclizing squalene-2,3-epoxide into cycloartenol, β-amyrin and α-amyrin in the presence of Triton X-100. These activities were efficiently solubilized by treatment with Triton X-100 and separated by chromatography on hydroxy apatite, DEAE-cellulose, isoelectric focusing and gel filtration. The purified cycloartenol cyclase showed a single band on SDS-polyacrylamide gel electrophoresis with Mr = 54000, while β-amyrin cyclase gave a single band with Mr = 28000.
Objective To study the best extracting technology of Compound Taizishen Granule prescription.Methods The total polysaccharides and total saponins were determined by UV spectrophotometer.With the content of total polysaccharides and total saponins as comprehensive indexes,orthogonal test was applied to investigation of the water addition,extraction time,and extraction times for optimizing the best extracting technology.Results The best extracting technology was using 10 times the amount of water,extracting 3 times and 1.5 h per time.Conclusion The extraction technology is safe and effective,which can be used as extraction for Compound Taizishen Granule prescription.