Reactivity of Pure Candida rugosa Lipase Isoenzymes (Lip1, Lip2, and Lip3) in Aqueous and Organic Media. Influence of the Isoenzymatic Profile on the Lipase Performance in Organic Media

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• 作者：Neus Ló ; pez ; Mar&iacute ; a A. Pernas ; Lorenzo M. Pastrana ; Antoni Sá ; nchez ; Francisco Valero ; and Mar&iacute ; a L. Rú ; a
• 刊名：Biotechnology Progress
• 出版年：2004
• 出版时间：February 2004
• 年：2004
• 卷：20
• 期：1
• 页码：65 - 73
• 全文大小：117K
• 年卷期：v.20,no.1(February 2004)
• ISSN：1520-6033

文摘

Three pure isoenzymes from Candida rugosa lipase (CRL: Lip1, Lip2, and Lip3) werecompared in terms of their stability and reactivity in both aqueous and organic media.The combined effect of temperature and pH on their stability was studied applying afactorial design. The analysis of the response surfaces indicated that Lip1 and Lip3have a similar stability, lower than that of Lip2. In aqueous media, Lip3 was themost active enzyme on the hydrolysis of p-nitrophenyl esters, whereas Lip1 showedthe highest activity on the hydrolysis of most assayed triacylglycerides. The highestdifferences among isoenzymes were found in the hydrolysis of triacylglycerides. Thus,a short, medium, and long acyl chain triacylglyceride was the preferred substrate forLip3, Lip1, and Lip2, respectively. In organic medium, Lip3 and Lip1 provided excellentresults in terms of enantioselectivity in the resolution of ibuprofen (EF value over0.90) and conversion, whereas initial esterification rate was higher for Lip3. However,the use of Lip2 resulted in lower values of conversion, enantiomeric excess, andenantioselectivity. In the case of trans-2-phenyl-1-cyclohexanol (TPCH) resolution,initial esterification rates were high except for Lip3, which also produced poor resultsin conversion and enantiomeric excess. The performance of the pure isoenzymes inthe enantioselectivity esterification of these substrates was compared with differentCRL crude preparations with known isoenzymatic content and the different resultscould not be explained by their isoenzymatic profile. Therefore, it can be concludedthat other factors can also affect the catalysis of CRL and only the reproducibilitybetween powders can ensure the reproducibility in synthesis reactions.