Preparation Of Immobilized Lipase From Ice Glue And Study On Catalytic Synthesis Of Fatty Acid Methyl Ester

The preparation of a new type of immobilized lipase has been studied in this paper. Then the immobilized lipase was used in the catalyzed synthesis of fatty acid methyl ester. It was specifically composed of the following three parts:1. The preparation and characterization of the immobilized lipaseAcrylamide, N, N-methylene double acrylamide, acrylic acid and allyl amine have been mixed together, added with the lipase, ammonium persulfate and sodium bisulfite have been used as initiators. Conventional gel immobilized lipase and ice gel immobilized lipase got by copolymerization at room temperature and-20degrees celsius respectively. Compared with the free enzyme and conventional gel immobilized lipase, we can find that frozen gel immobilized lipase has a higher catalytic activity and shows good stability. The pH of the polymer and the ratio of monomer have also been discussed. The experimental results show that neutral, with the mass ratio of acrylamide and N, N-methylene double acrylamide10:1are the best synthetic conditions for polymer. Then the prepared immobilized lipase was characterized with scanning electron microscopy (SEM). Finally, the best reaction temperature was chosen at40℃, the best alcohol oil ratio was chosen1:1, and the quality percentage of the immobilized lipase and oil was chosen20%.2. Analysis of the process of three glyceride ester exchange with micellar electric capillary chromatographyThe esterification reaction of ester compounds was catalyzed by the prepared immobilized lipase. And ester exchange reaction of ester compounds of the product are analyzed with the micellar electric capillary chromatography. The best condition of micelle electric capillary chromatographic analysis has been obtained by the experiment.234nm was chosen as the best absorption wavelength,20s was chosen as the optimal sampling time with elevation difference sampling method,-5kV was chosen as the best separation voltage with the current of about50mA. The buffer is composed of SDS with concentration of115mmol/L (3.31%), borax with concentration of10mmol/L (92.5%),1.2%normal octane and3%normal butanol with pH of8.0. On this condition, glycerol trioleate and glycerol trioleate catalyzed by the immobilized lipase for a period of time was determined with micellar electric capillary chromatography respectively. Different chromatograms of ester exchange reaction have been obtained. It can be embodied that the immobilized lipase has good catalytic performance for ester compounds.3. Synthesis of fatty acid methyl ester catalyzed by immobilized enzymeWith the combination of the microemulsion capillary chromatography, the prepared immobilized lipase was used for further application. Then different cooking oil was catalyzed by the immobilized lipase and different chromatograms can be obtained. The experimental results show that the prepared immobilized lipase has a certain catalytic effect on different cooking oil. It can be applied in catalyzing cooking oil from different sources. Then cooking oil was studied to synthesize fatty acid methyl ester catalyzed by the prepared immobilized lipase. Then the influence of different experimental conditions (the catalytic reaction time, reaction temperature) on the cooking oil catalytic conversion has also been discussed. Finally,24h was chosen as the best catalytic time,40℃was chosen as the best catalytic reaction temperature. The experiments show that the immobilized lipase not only has good catalytic performance for different oil, but also has a high stability. So presumably the prepared immobilized lipase will have a broad application prospect.