The Immobilization Of PEL And Components Analysis Of Soybean Oil Hydrolysis Product

The hydrolysis of soybean oil catalyzed by Penicillium Expansum Lipasewas studied for preparation of natural anti-bubbing agent in this work. The assayof lipase activity was improved and the immobilization method of lipase wasconfirmed. Furthermore, the assay of quantitive determination the component ofsoybean oil was studied. Based on the systemic research above, the reactionconditions of the hydrolysis of soybean oil were screened out. All the work willprovide the theory foundation for the exploration of soybean oil. In assays of determination lipase activity, the stability of the data withtributyrin and triolein (obtained from Sigma) as substrates was better than that ofthe data with olive oil (obtained from China) as substrate. It could be found thatthe lipase activity determined with triolein as substrate was about 2-3 times asmuch as the results with olive oil and tributyrin as substrates. Compared several frequently-used assays of determination lipase activity,it's found that pre-emulsification of the substrate and selection of a correctdetermination method could give a stable lipase activity data. The method ofdetermination lipase activity improved in this work could be applied widelybecause the assay conditions were simple and the operation was easy. In immobilization research of lipase, diatomaceous earth was used asprotein carrier. The results of the optimal conditions for lipase immobilizationwere that pH is 7.7; temperature range is from 30℃ to 35℃; ionic strengthrange is from 0.01mol/Lto 0.03mol/L; the optimal ratio of diatomite and lipase is8:1. Under the optimal conditions, the protein amount absorbed on thediatomaceous earth was 52mg/g and the lipase activity was 1301 μmol·min-1·g-1.The immobilized lipase activity in hydrolysis of soybean oil was 43吉林大学硕士学位毕业论文 高 贵remained 1137μmol·min-1·g-1 even after 3 times reused. In hydrolysis of soybean oil, the optimal conditions were screened out. Thereaction system was 1:2(oil/buffer=v/v); the lipase was 0.25g/20mL(160u/mL)in buffer,the reaction time was 2.5h and the stir speed was 160rpm. In this work, the analysis method of hydrolysis product component has alsobeen improved. TLC was used to segregate the different glyceride and fatty acidin hydrolysis product components and to analyze the different glyceride and fattyacid qualitatively. The silica gel column was used to obtain different components.The different components in hydrolysis product were distinguished byHPLC-MS. The acid number of hydrolysis product has also been determined.Through comparison and analysis, it could be found that the relative content ofdifferent components in hydrolysis product possessed some certain relativity withthe acid number. So the acid number could be used as a indicator to estimate therelative content of different components in hydrolysis product.