Mutation Breeding Of High Oil Content Chlorella And Its Grease Extraction

The microalgae oil is an important renewable resources, similar with vegetable fats and oils, can be used in the production of bio-diesel, and which is expected to become a golden key to unlock the energy crisis"in post-oil era". In recent years, the study of biodiesel production from microalgae is listed as one of the key projects of the Ministry of Science and Technology "973" and "863" plan with causing our government, and the grape algae(Botryococcus braunii) and Chlorella (Chlorella pyrenoidosa) are main oil-producing algae. Chlorella is considered to be one of the most potential algae for industrial production of biodiesel because biodiesel of Chlorella as raw materials is very appropriate for the combustion of the diesel engine. the high oil content and rapid growth characteristics of chlorella. The purpose of this study is to develop a microalgae of high fat content as a resources to prepare biodiesel, and mainly focused on the rapid method for biomass and fat content determination of Chlorella, biomass and fat content changes of chlorella in cultivation cycle, the comparison of different extraction methods of Chlorella oil using analysis of gas chromatography, screening Chlorella algae species with high fat content from UV, EMS. EMS composite cerulenin mutagenesis. The results are as follows:1. There is an good linear relationship between the absorbance values of684nm and cell density and dry weight of chlorella.and the OD value can quickly converted by cell density and dry weight of the algae. There is an ideal linear relationship between the absorbance values of423nm and the reactant of different content fat. R2>0.999, to made the standard curve between the fat content and the OD value,, and can converse fat content from OD value of reactants.2. The biomass of Chlorella accumulated to maximum when growth to a stable period which similar with fat content. Therefore, the best time of the harvest is stabilization period (10-11d)for chlorella.3. The different lipin yield, different ratio of long-chain fatty acids(C14-C22) to total fatt y acids and different extract purity of fatty acid were obtained by the different methods o f extraction. And, the method of chloroform-methanol extraction is a simple and effici ent method of lipin extraction,with lipin yield of22.40±0.89%and85.84%long chain fatt y acids(C14-C22); In Soxhlet extraction,we get the highest yield rate of24.37±0.21%with Anhydrous alcohol as an extraction solvent, and get up to10kinds of fatty acids with petr oleum ether as an extraction solvent.4. The positive mutation rate was16.7%in UV mutagenesis of Chlorella, and the fat content was increased5.63%than that of the original strain. The positive mutation rate was40%in EMS mutagenesis of Chlorella, and the highest fat content was increased7.29%than that of the original strain. The positive mutation rate was60%in EMS compound cerulenin mutagenesis of Chlorella, and the highest fat content was increased9.03%than that of the original strain. The biomass of the alga in UV and EMS mutagenesis did not significantly improve, but which has slightly improved.in the EMS compound cerulenin mutagenesis...