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Enhancing the thermal stability of lipases through mutagenesis and immobilization on zeolites

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Abstract

The hydrolysis reaction of p-nitrophenyl butyrate catalyzed by lipases was followed with in situ UV/vis diode array spectrophotometry. Five enzymes - Candida antarctica lipase B and Fusarium solani pisi cutinase wild-type and three single-mutation variants - were tested as catalysts in homogeneous conditions and immobilized on zeolite NaY, on a polyacrylate support and as cross-linked aggregates. Using deconvolution techniques and kinetic modeling, the thermal stability of the different biocatalysts was compared in operational conditions and the results were supported by steady-state enzyme fluorescence measurements. We concluded that both the mutagenesis and the immobilization on zeolite NaY had a positive effect on the thermal stability of F. solani pisi cutinase.

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Acknowledgments

The authors gratefully thank Novozymes A/S (Denmark) for the kind gifts of Novozym® 435 and Lipozyme® CaLB L. L. Costa and V. Brissos acknowledge Ph.D. grants (SFRH/BD/19108/2004 and SFRH/BD/9019/2002) from Fundação para a Ciência e Tecnologia.

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Authors and Affiliations

  1. IBB-Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisbon, Portugal

    L. Costa, V. Brissos, F. Lemos, F. Ramôa Ribeiro & J. M. S. Cabral

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  1. L. Costa
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  2. V. Brissos
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  3. F. Lemos
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  4. F. Ramôa Ribeiro
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  5. J. M. S. Cabral
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Correspondence to F. Lemos.

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Costa, L., Brissos, V., Lemos, F. et al. Enhancing the thermal stability of lipases through mutagenesis and immobilization on zeolites. Bioprocess Biosyst Eng 32, 53–61 (2009). https://doi.org/10.1007/s00449-008-0220-x

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  • DOI: https://doi.org/10.1007/s00449-008-0220-x

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