Study On The Tissue And Suspension Cell Culture Of Gardenia Jasminoides Eills And The Isolation, Purification And Biological Function Of Its Polysaccharides

The research on Polysaccharide is very important in the field of biochemistry, and it is becoming a new hot point. There are many reports on plant polysaccharide recently, especially those of traditional Chinese medicine. The authors found polysaccharide has many biological activities, such as anti-tumor, anti-virus, anti-bacteria and anti-oxidize etc. Part of biological activities, isolation, purification and feasibility of producing GJEP by biotechnology were studied here.Gardenia jasminoides Bills is a kind of traditional Chinese medicine. The compositions used of traditional Chinese medicine are those that can dissolve in water. GJEP is one of them, it is important to restrain the growth of tumors and anti-oxidation .In vitro, 100ug/ml GJEP can restrain sarcoma 180 cell (58%), Hca-f ascites hepatoma cell (51%) and man erythroleukemia K562 cancer cell (63%). The inhibition ratio of GJEP on Hca-f ascites hepatoma in mice achieved 49%, and the results of ig are better than those of ip.GJEP can scavenge super-oxygen negative ion free radical, and the scavenging ratio is 83.48%. However, scavenging ratio of GJEP on hydroxyl free radical is 22.31%. Some medal ion can influence the scavenging ratio of GJEP on free radical. Zn2+,Fe2+ and Ca2+ can decrease the capacity of GJEP on scavenging ?O2- in varying degrees, Cu2+ can increase a litter the capacity of GJEP on scavenging ?O2- .However, these four medal ions all can increase the effect on scavenging ?OH. GJEP can increase SOD enzyme activities and decrease MDA content of mice, the changes are related with the growth of tumor. The mice that have Dowerful SOD activity, have low MDA content, their tumors are heavy; On the contrary, the mice that have weak SOD activity, have highMDA content, their tumors are light. So we infer that GJEP can restrain the growth of tumor by increasing SOD enzyme activities of mice to scavenge unnecessary free radical.The production of GJEP was studied by suspension cell culture. The results show that:(l) The size of cell aggregate does not affect the production of GJEP. There are some structure for transferring material in cell aggregate; (2) The subculture periods of 5-10 days are good for keeping the capacities of cell growth and polysaccharide production; (3) The inoculum concentration of 80g/L cell can shorten the time, which the cell biomass achieve the biggest value needed; (4) Sucrose can be decomposed into glucose and fructose in suspension culture. The cell of Gardenia jasminoides Bills has priority to using glucose; (5) The capacity of GJEP production was decreased in later stage, one of the important reason was that the carbon source in the medium was used up; (6) Excessive nitrogen source was not good for producing GJEP; (7) Auxins have more affective on cell growth and polysaccharide production. The result of 2,4-D is priority to that of NAA and the result of NAA is priority to that of IAA; (8) Gardenia jasminoides Bills cell is not sensitive to shearing power, cell can grow normally in 80~130r/min rotational speed of table; (9) Red light and blue light are two important lights on cell growth and polysaccharide production in suspension culture of Gardenia jasminoides Bills; (10) The effects of temperature and PH value on cell growth and polysaccharide production of GJEP are the same as other plants.By selecting high production cell of GJEP and changing medium and cultural condition, growth of cell and polysaccharide content were increased. The suspension culture of Gardenia jasminoides Bills has some special character, but there is a regular that if the condition is good for cell growth, then it is probably not good for biological synthesis of GJEP.AbstractUsually, GJEP is produced at the early stage of suspension cell culture, and the yellow pigment of Gardenia jasminoides Bills is produced from the 12th day of suspension cell culture, so we can remove the disturbance of yellow pigment from GJEP by controlling the culture conditions and harvest time of cell, that is, inhibiting the synt...