Effects Of Precursor And Inducer On Biomass Accumulation Of Suspension Cells And Content Of Baicalin From Scutellaria Baicalensis

Scutellaria baicalensis, as a traditional Chinese medicine, has a long history in China. S. baicalensis was paid more attention by biological and medical community in recent years. The active components of baicalin was paid more and more attention. Suspension culture technology was applied to improve the content of secondary metabolite for S. baicalensis. The effects of the combination of inculation amount, carbon source, medium p H, the combinations of 6-BA and NAA on biomass of suspension cultures and baicalin contents from Scutellaria baicalensis were analyzed by using combination of single factor and uniform design, and the effects of precursor and elicitor on the biomass of the suspension cultures and the accumulation of baicalin were studied. The changes of physiogical and biochemical factors in culture process were determined, and relationships between enzyme spectrum of POD & PAL activities of the suspension cultures from Scutellaria baicalensis and baicalin contents were analyzed. The following points are the theoretical foundation for the large-scale commercial process. The results showed as the following:(1) Through the analysis of experimental results in different physical and chemical factors, the inoculum size of S. baicalensis is 1.0 g per bottle by suspension cultures; the optimal carbon source is sucrose, the sucrose concentration being 3%,which is the most suitable for growth of suspension cell lines; the optimal p H value which is fit for the growth of suspension cell line and the accumulation of metabolite is 5.8. The combination of 0.2 mg·L-1's 6-BA and 0.1 mg·L-1's NAA is optimal for the growth of suspension cell line.(2) Relationship of the dry/fresh weight, the p H value of the medium, conductivity, activity of PAL & activity of peroxidase with content of S. baicalensis were analyzed respectively through SPSS Statistics; the dry/fresh weight, conductivity & activity of peroxidase and content of baicalin appear significant positive correlation.(3) In the orthogonal experiment of precursor & elicitor, result of range analysis shows that : the fresh weight is the highest while adding 0.4 mg·L-1 yeast extract and 8 mg·L-1 phenylalanine(Phe) on the 12 th day;the dry weight is the highest while adding 0.4 mg·L-1 yeast extract and 6 mg·L-1 Phe on the 12 th day; the activity of phenylalnine ammonialyase(PAL) of suspension cell line for S. baicalensis is the highest while adding 0.8 mg·L-1 yeast extract on the 8th day and 6 mg·L-1 Phe on the 6th day;the activity of peroxidase of suspension cell line for S. baicalensis is the highest while adding 0.4 mg·L-1 yeast extract on the 12 th day and 6mg·L-1 Phe on the 9th day; the content of baicalin for suspension cell line is the highest while adding 0.6 mg·L-1 yeast extract on the 12 th day and 6 mg·L-1 Phe on the 9th day. According to the analysis of the variance, it's optimal for the growth of the fresh weight for suspension cell line of S. baicalensis while adding yeast extract on the 6th day, and the effect of other factors are not significant. It can be seen from the peroxidase isozyme spectrum, the activity of the Phe is the strongest while adding 0.8 mg·L-1 yeast extract on the 6th day and 4 mg·L-1 Phe on the 9th day.