Cell Suspension Culture Of Glycyrrhiza Uralensis Fisch In Bubble Bioreactor And Analisis Of Component And Activity

Glycyrrhiza uralensis Fisch, which belongs to the Leguminosae family is a perennial herbaceous plant. It is a common Chinese herb and has already been widely used in many fields, such as medicine, food and cosmetic. But due to enormous utilization of glycyrrhiza, the resource has become scarce in the world. In this paper we have studied the glycyrrhiza cell in bubble bioreactor, in order to product the active component through cell culture technology, which will remit lacking of glycyrrhiza resources to some extent.The culture conditions of glycyrrhiza cell was optimized in bioreactor, included, investigating consuming of carbon source which was the main energy for cell growth; analyzing the effect of replenish carbon source for the growth and active component accumulating of glycyrrhiza cell; observing the addition amount of phenylalanine (PHE) and Methyl jasmonate (MJ), which were a flavonoid precursor and an elicitor for promoting active component in glycyrrhiza, respectively; also analyzing the effect of PHE and MJ for growth and active component accumulating of glycyrrhiza cell. Results showed that both of PHE and MJ could enhance the content of total flavonoid and glycyrrhiza polysaccharide in glycyrrhiza cell, and they could also slightly inhibit the cell growth.Compared with the fed-batch, two-stage cultivation could obviously promote the production of flavonoids and polysaccharide in a 5-L bubble bioreactor. It was demonstrated that when using two-stage cultivation the dry cell weight were 1.05-fold and 1.15-fold greater than fed-batch cultivation and batch cultivation, respectively in the 15th d; also the contents of polysaccharide were increased by 1.14-fold and 2.12-fold when compared to fed-batch cultivation and batch cultivation respectively. In addition, the two-stage cultivation could obviously enhance total flavonoid in glycyrrhiza cell, and it reached to 132.36 mg in 15th d, which was 2.26-fold and 2.67-fold greater than the fed-batch cultivation and batch cultivation, respectively. Through the contrastive analysis difference between the glycyrrhiza cell and wild glycyrrhiza in their components, using eight standards which included two saponins and six flavonoids. In was found that the contents of glycyrrhizin, chalcone A and isoliquiritigenin in glycyrrhiza cell were 2.42-fold,1.59-fold and 1.69-fold higher than that in wild glycyrrhiza. In the bioactivities comparison of glycyrrhiza cell and wild glycyrrhiza, Two inflammation models (ovalbumin-induced paw edema in rat and xylene-induced ear edema in mice) was chose to observe that the in inflammatory activity of glycyrrhiza cell was greater than that of wild glycyrrhiza. Finally, in order to investigate the anti-inflammatory mechanism of glycyrrhiza flavonoid, the enzyme activity of SOD was measured in tissues.