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Optimization in the immobilization of penicillin G acylase by entrapment in xerogel particles with magnetic properties

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Abstract

Biocatalysis presents a sound alternative to chemical synthesis in the field of drug production, given the highly selective nature of biological catalysts. Penicillin G Acylase (PGA) from E. coli is currently used to hydrolyze penicillin G (PG) and catalyzes the synthesis of β-lactam antibiotics. In this work, particular emphasis is given to recent developments in penicillin G acylase immobilization, by entrapment simultaneously with nano-magnetic particles in a silica matrix. The sol–gel biocatalytic particles were prepared either by a conventional method (crushed powder) or by a more recent approach, based in an emulsion system using 150 mM AOT/isooctane, which allowed for the formation of spherical micro- and nanobeads. The effects on PGA activity of different sol–gel precursors, additives, enzyme concentration, aging, drying conditions and mechanical stability were evaluated. After these optimization studies, a mechanically stable carrier based on porous xerogels silica matrixes, starting from tetramethoxysilane (TMOS) with 65–67% PGA activity yield in these carriers allowed an immobilization yield of 74 mg protein g −1dry sol–gel and 930 Ug −1dry sol–gel for specific activity were obtained.

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Acknowledgments

S. M. S. A. Bernardino, N. I. Estrela and P. Fernandes acknowledge Fundação para a Ciência e a Tecnologia (Portugal) for financial support in the form of the PhD grants SFRH/BD/30632/2006, SFRH/BD/18639/2004 and for a contract under Program Ciência 2007, respectively.

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Authors and Affiliations

  1. ESTM—School of Tourism and Maritime Technology, Marine Resources Research Group, Polytechnic Institute of Leiria, 2520-641, Peniche, Portugal

    Susana Bernardino

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

    Susana Bernardino, Nídia Estrela, Vanessa Ochoa-Mendes, Pedro Fernandes & Luís P. Fonseca

  3. Department of Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisbon, Portugal

    Susana Bernardino, Nídia Estrela, Vanessa Ochoa-Mendes, Pedro Fernandes & Luís P. Fonseca

  4. IBB—Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine, Universidade do Algarve, 8005-139, Faro, Portugal

    Nídia Estrela

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  1. Susana Bernardino
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  2. Nídia Estrela
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Correspondence to Susana Bernardino.

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Bernardino, S., Estrela, N., Ochoa-Mendes, V. et al. Optimization in the immobilization of penicillin G acylase by entrapment in xerogel particles with magnetic properties. J Sol-Gel Sci Technol 58, 545–556 (2011). https://doi.org/10.1007/s10971-011-2426-7

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