大孔树脂固定化脂肪酶及在微水相中催化合成生物柴油的研究

以不同大孔树脂吸附法固定化假丝酵母99_125脂肪酶,在微水有机相中的应用表明非极性树脂NKA是最佳的固定化载体。分别以正庚烷及磷酸盐缓冲液作为固定化介质,发现在正庚烷介质中树脂NKA的固定化效率能够达到98.98%,与采用磷酸盐缓冲液作为介质相比,固定化酶的水解活力和表观酶活回收率分别提高了4.07和3.43倍。考察了在微水相中固定化酶催化合成生物柴油的催化性能,结果表明,在给酶量为1.92∶1(初始酶粉与树脂的质量比),pH值为7.4,体系水含量为15%(水与油的质量比),反应温度为40℃条件下,固定化酶具有最佳的催化能力;以正庚烷为介质固定化脂肪酶催化合成生物柴油,采用三次流加甲醇的方式,单批转化率最高达到97.3%,连续反应19批以后转化率仍保持为70.2%。

Immobilization of Lipase on Macroporous Resin and its Application in Synthesis of Biodiesel in Low Aqueous Media

Lipase from Candida sp. 99-125 was immobilized by physical adsorption onto macroporous resins. The results showed that the nonpolar resin NKA was the best carrier used in low aqueous media. 98.98% of degree of immobilization can be achieved when the adsorption procedure was performed in the presence of heptane. The hydrolytic activity and the apparent activity recovery of lipase adsorbed on resin in heptane was 4.07 and 3.43 times higher than that of lipase adsorbed in sodium phosphate buffer, respectively. The catalytic properties of immobilized lipase for production of biodiesel in low aqueous media were studied. Immobilized lipase displayed the highest activity when the crude enzyme/resin weight ratio was 1.92:1 and the water content(water/oil weight ratio) was 15% at 40 degrees C under pH 7.4. As lipase was adsorbed on NKA in heptane to produce biodiesel, the batch conversion rate can reach 97.3% when a three-step methanolysis protocol was used. After 19 consecutive batches, the conversion rate remained 70.2%.