Kinetic resolution of drug intermediates catalyzed by lipase B from Candida antarctica immobilized on immobead‐350 |
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| Authors: | Maísa Pessoa Pinheiro Nathalia Saraiva Rios Thiago de S Fonseca Francisco de Aquino Bezerra Enrique Rodríguez‐Castellón Roberto Fernandez‐Lafuente Marcos Carlos de Mattos José C S dos Santos Luciana R B Gonçalves |
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| Affiliation: | 1. Dept. de Engenharia Química, Universidade Federal do Ceará, CEP 60455‐760, Fortaleza, CE, Brazil;2. Dept. de Química Organica e Inorganica, Laboratório de Biotecnologia e Síntese Organica (LABS), Universidade Federal do Ceará, Campus do Pici, Fortaleza, CE, Brazil;3. Dept. de Química Inorgánica, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, Boulevard Louis Pasteur, Málaga, Spain;4. Dept. de biocatalisis, ICP‐CSIC, Campus UAM‐CSIC, Cantoblanco, Madrid, Spain;5. Departament of Chemical Engineering, Instituto de Engenharias e Desenvolvimento Sustentável, Universidade da Integra??o Internacional da Lusofonia Afro‐Brasileira, CEP 62785‐000, Acarape, CE, Brazil;6. Dept. de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, CEP 60455‐760, Fortaleza, CE, Brazil |
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| Abstract: | Novozyme 435, which is a commercial immobilized lipase B from Candida antarctica (CALB), has been proven to be inadequate for the kinetic resolution of rac‐indanyl acetate. As it has been previously described that different immobilization protocols may greatly alter lipase features, in this work, CALB was covalently immobilized on epoxy Immobead‐350 (IB‐350) and on glyoxyl‐agarose to ascertain if better kinetic resolution would result. Afterwards, all CALB biocatalysts were utilized in the hydrolytic resolution of rac‐indanyl acetate and rac‐(chloromethyl)‐2‐(o‐methoxyphenoxy) ethyl acetate. After optimization of the immobilization protocol on IB‐350, its loading capacity was 150 mg protein/g dried support. Furthermore, the CALB‐IB‐350 thermal and solvent stabilities were higher than that of the soluble enzyme (e.g., by a 14‐fold factor at pH 5–70°C and by a 11‐fold factor in dioxane 30%–65°C) and that of the glyoxyl‐agarose‐CALB (e.g., by a 12‐fold factor at pH 10–50°C and by a 21‐fold factor in dioxane 30%–65°C). The CALB‐IB‐350 preparation (with 98% immobilization yield and activity versus p‐nitrophenyl butyrate of 6.26 ± 0.2 U/g) was used in the hydrolysis of rac‐indanyl acetate using a biocatalyst/substrate ratio of 2:1 and a pH value of 7.0 at 30°C for 24 h. The conversion obtained was 48% and the enantiomeric excess of the product (e.e.p) was 97%. These values were much higher than the ones obtained with Novozyme 435, 13% and 26% of conversion and e.e.p, respectively. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:878–889, 2018 |
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| Keywords: | lipase immobilization– stabilization epoxy‐support covalent immobilization rac‐indanyl acetate rac‐(chloromethyl)‐2‐(o‐methoxyphenoxy)ethyl acetate lipase B from Candida antarctica |
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