The Study Of The Fermentation Tank Process Of Nocardia Rubra And The Further Study Of Antitumor Pharmacology

The Nocardia Rubra belongs to Nocardia. Rhodococcus, Corynebacterium and Mycobacterium make up a distinctive suprageneric taxon-the mycolata because of the similarity of their cell envelope composition and organization which are different from other Gram-positive bacterias. The cell envelope of Mycolata are dominated by an arabinogalactan cell wall polysaccharide,large branched-chain fatty acids and mycolic acids, Which are covalently assembled into a peptidoglycan-arabinogalactan-mycolic acid matrix(MAPc). And the MAPc is confirmed to a immunomodulator. Clinically, N-CWS has good curative effects on malignant tumors, cervical erosion and better reconstruction.The optimized culture conditions were got by single factor experiments. And the response surface methodology(RSM) was adopted to optimize the three important fermentation conditions in 5L fermentor. The predicted maximum biomass was 26.64g/L which was close to the experimentally measured biomass. The results showed that RSM was feasible to optimize the fermentation conditions in 5L fermentor of Nocardia rubra, improved 9.8% compared with the 250 ml shake-flask.Based on the experimental data resulted from 5L batch fermentation tank, the batch fermentation kinetics of Nocardia rubra in 5L fermentor was studied, Mathematic models of Nocardia rubra mycelia growth and substrate consumption were established on the basis of Logistic and Boltamann equations respectively.The optimized conditions of adding glucose was got based on the DO-stat at 10%: the initial concentration was 20g/L, the optimized fermentation time was 36h, the optimized concentration of the additional glucose was 50g/467ml and the best additional time was one. Compared the DO-stat fed-batch fermentation with batch fermentation, the biors Nocardia rubrawas improved 43.91 %.The disruption of Nocardia rubra cell wall by high pressure hoxnogenizer was studied. The effect of different conditions such as pressure, cycle number and temperature were investigated through single factor experiments. Preliminary results showed that the optimized condition was 2250 bar, about 25°C and 3 cycle numbers. The oral experiment showed that the different dosages of debris of Nocardia Rubra cells had different inhibitory influence on mouse. The low dose, middle dose and high dose reached 23.96%、44.38% and 60.65% respectively. And this result was dose-dependent. The thymus index, spleen index and the phagocytic activity of peritoneal macrophage proveed that the debris of Nocardia Rubra cells can increased the nonspecific immunity of mouse. Simultaneously the activities of the acid phosphatase inside the peritoneal macrophage was increased along with the dosages with also showed a dose-dependent way.This study analyzed antitumor active ingredients of N-CWS, and the PG was found to be the material base to induce the apoptosis of HepG2. The Wright Giemsa stain solution was used to examine the morphas of HepG2 cells which confirmed the last conclusion.