The Technics Of Using Wastewater From Cassava Ethanol Production To Culture Chlorella Pyrenoidosa And The Bioconversion Of Chlorella Pyrenoidosa By Ganoderma Lucidum

The dissertation was focused on the technics of using wastewater from cassava ethanol production to culture Chlorella pyrenoidosa, the determination of operation conditions and technological parameters, aimed to recycle the treated wastewater in ethanol fermentation to save energy, realize depurated fermentation, enhance economical benefit. In order to increase the applied value of C. pyrenoidosa, through anti-tumor experiment in vivo, the medicine effect of bioconversion ferment liquid was estimated after C. pyrenoidosa was bio-converted by Ganoderma lucidum. And the increased components of ferment liquid were also studied.After Spirulina platensis, Chactoceros muelleri, Anabaena sp., Scenedesmus obliquus and C. pyrenoidosa were cultured in 250-ml conical flasks, with wastewater from cassava ethanol production of different concentration (from 10 to 100%), C. pyrenoidosa was the optimum algae cultured in the undiluted wastewater through selection. The results showed that the optimum cultivation conditions were initial pH of 6.0, temperature at 27oC,continuous illumination at 3000 lux and cycle speed of 110 ml min-1 in a cycle tubular photobioreactor, when the undiluted wastewater was used to culture the algae..Batch, fed-batch and continuous culture was operated and researched in the cycle tubular photobioreactor under above optimum conditions. After the logarithmic phase of batch operation with wastewater of pH 6.0, the reactor could be continuously operated with pH natural (3.8) wastewater as feed solution. By a dilution ratio of 0.17 day-1, it could be operated stably for over 30 days in continuous operation. pH, removal rate of chemical oxygen demand and biomass (cell dried weight) concentration ranged from 6.22 to 6.47, 72.21 to 76.32 % and 3.55 to 3.73 g l-1, respectively. The removal rate of lactic acid, acetic acid, citric acid and succin acid in the wastewater were 96.42 %, 100 %, 95.89% and 39.44 %, respectively. And the algae which have high value were harvested. After treatment, the wastewater could be used again in the process of ethanol fermentation.C. pyrenoidosa powder was appended by different concentration into the basic medium of G. lucidum, the G. lucidum fermentation was studied. The optimum concentration was 10 g l-1, and the output of polysaccharides and other bioactivity matters in ferment liquid was enhanced significantly. The ethanol extraction from C. pyrenoidosa, from ferment liquid of G. lucidum in basic madium, from ferment liquid of bioconverted C. pyrenoidosa by G. lucidum were contrasted and analyzed by HPLC. The changes of component kinds and their concentration illustrated that G. lucidum could bio-broke the cell wall of C. pyrenoidosa and bioconversion was achieved. The high dose group (13 ml kg-1d-1) of bioconversion ferment liquid had the highest tumor inhibition (67.70%) which was higher than that of ferment liquid with basic media(41.61%) and that of mixture liquid(43.48%)contained ferment liquid with basic media and water extraction of C. pyrenoidosa at same concentration, significantly. The result showed 1+1>2.The bioconverted polysaccharides had the best activity of anti-tumor and immunological function. The anti-tumor inhibition was 76.2% at the optimum dose (0.2 g kg-1d-1). The polysaccharides from ferment liquid of bioconverted C. pyrenoidosa by G. lucidum had higher medicine effect not only than polysaccharides from ferment liquid of G. lucidum in basic madium, from C. pyrenoidosa, but also than the mixed polysaccharides. The increase of bioconversion polysaccharides in ferment liquid and the increase of its tumor inhibition was one of the reasons which resulted in the increase of anti-tumor of bioconversion ferment liquid.The increase of component whose retention time was 25.778 min was high. The component might play an important role and advanced action in the bioactivity of conversation ferment liquid, was separated from the crude butanol extraction of culture liquid and purified by macroporous resin HPD-400 and octadecyl silane column. It was likely conjectured as a flavone according to UV, HPLC/MS and IR. Flavone compound usually had anti-tumor and other bioactivity. The increase of flavone in ferment liquid was also one of the reasons which resulted in the increase of anti-tumor of bioconversion ferment liquid.