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Four different commercial lipases of Pseudomonas spp (PS-P), Candida rugosa (CR-P), Mucor javanicus (MJ-P) and Rhizopus arrhizus (,RA-P) have been immobilized by adsorption with celite (PS-A, CR-A, MJ-A, RA-A) and by entrappment with hydrophobic silane (PS-E, CR-E, MJ-E, RA-A). The conversion rate of capronic acid and ethanol to ethyl caproate of thase free and immobilized lipase in organic solvent n-hexane, in aqueous alcohol solution, and in the supercritical fluid of carbon dioxide system have been measured and compared in this study. The free PS lipase (PS-P) is found to have the highest esterification activity in n-hexane. The conversion rate can reach 80% after reaction for 8h. Each mg o f PS-P can synthesize 0.0462 μM of ethyl caproate per minute. The esterification activity of lipase is increased greatly after immobilization, especially for the entrapped PS lipase (PS-E). The conversion rate of PS-E can reach 90% at 1.5h, and each gram of PS-E can synthesize 298.3 μM of ethyl caproate per min. The optimum enzyme dose for adsorbed lipase is 400mg/g of celite, but for entrappment in hydrophobic silane is more than 800mg. Adding trace amount of water in n-hexane can increase the esterification activity of celite adsorbed lipase up to water content to 0.0625%. Among these four lipase, only PS lipase show some esterification activity in 40% alcohol solution. The conversion rate of caproic acid to ethyl caproate for PS-P, PS-A and PS-E are 15.4%, 23% and 40% respectively. The ethyl caproate synthesized by PS-E is more stable than PS-P and PS-A, and it''s rate of ester formation is increased with the increasing in alcohol content. In 60% alcohol solution, the conversion rate for PS-E at 5h is 43%. The immobilized lipases have been used repeatly batchwise for ethyl caproate synthesis in n-hexane or in 40% alcohol solution. For celite adsorbed lipase (PS-A), more than half enzyme activity is lost at the second batch no matter how is the polarity of the reaction medium. But for the hydrophobic silane entrapped PS lipase (PS-E), the half life of enzyme activity in n-hexane is 40 batches and in 40% alcohol solution is 16 batches. The immobilized lipases have also been applied in the supercritical fluid of carbon dioxide system for ethyl caproate synthesis. Better results can be obtained for the entrapped PS lipase (PS-E, the conversion rate can be got to 80% after starting for 8h and maintained at 50% when reaction proceed for 50h.
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