The Preparation Of Carica Papaya Lipase And Its Catalytic Resolution On Flurbiprofen Methyl Ester

The Carica papayal as a kind of agricultural and sideline products. It had good market development and scientific value. It is a kind of food additive, and has the function of weight loss. The Carica papayal lipase derived from a plant, had the very high biological catalytic function and low cost. The main contents of the thesis are as follows:(1) Firstly, Carica papaya lipase has been purificated and analyzed for hydrolysis activity, 13 percent of Carica papaya lipase was obtained from crude papain by its natural immobilized properties. The activities of lipase was measured, it was 90U/g. It had the highest hydrolysis activity during lyophilization in 8 hours. The highest hydrolysis activity of Carica papaya lipase was obtained at 35"C, and it also had higher hydrolysis activity at 30-4CTC, it had high enzyme activity at the pH 8.0, and the lipase was also very stable at pH6-10; the lipase had high thermal stability, it was stored at different temperature for 6 hours, the hydrolysis activity loss is small. And it had a good tolerance in the low polar organic solvents.(2) In the non-aqueous reaction system, the effects of the lipase content, buffer solution content, temperature, stirring speed and reaction time on the Carica papaya lipase catalyzed resolution reaction were investigated by the single factor experiments. The optimal conditions was built when the reaction was carried out in the isooctane with 10mg/mL of flurbiprofen methyl ester,70mg of Carica papaya lipase,5% of the buffer at150rpm and 35? for 168 hours. The highest conversion was 14%, and the enantiomeric excess of product was 87%.(3) Because the enantiomeric excess of product was lower by Carica papaya lipase catalyzed resolution, a (R)-preference lipase and a (S)-preference lipase catalyzed double kinetic resolution strategy has been designed. The (S)-flurbiprofen methyl ester was enriched using immobilized Novozym 435 lipase catalyzed hydrolysis and then was hydrolyzed selectively to (S)-flurbiprofen by Carica papaya lipase in microaqueous-isooctane biphase system. In this way, the double resolution was better than a single lipase catalyzed resolution, the enantiomeric excess of product was 91%.