Abstract
The bacterium Pseudomonas fluorescens 07A produces a protease with potential for industrial application. In order to remedy problems associated with the use of free enzymes and allow its reuse, the protease was immobilized on DEAE Sephacel® resin via three different strategies based on ionic interaction and covalent bonding. The matrix-bound enzymes were characterized in relation to their activity (pH, temperature and stability), reuse and storage. Immobilization raised the optimum temperature of activity from 37 °C to 50 °C, whereas the pH of highest activity changed from 7.5 to 7.0 or 8.0, depending on the immobilization strategy. Immobilization proved to be efficient for successive reuses, even leading to an increase in the enzymatic activity along the use. The immobilized enzyme also presented greater stability to high temperatures and storage conditions and has potential as a biocatalyst for industrial applications due to its high efficiency, stability and easy recovery.
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We thank the agencies Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), finance code 001 and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the continuous support to research in Brazil.
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KFC and JSM planned and performed the experiments, analyzed the data and wrote the manuscript. They contributed equally to the work and should be regarded as co-first authors. JAP contributed to the execution of the experiments and to writing the manuscript. TPC contributed to writing the manuscript and MRE planned and supervised the experiments and revised the manuscript.
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Cardoso, K.F., Miranda, J.S., de Paula, J.A. et al. Enhancement of the activity, stability and reusability of an extracellular protease from Pseudomonas fluorescens 07A via three different strategies of immobilization. Braz. J. Chem. Eng. 37, 485–493 (2020). https://doi.org/10.1007/s43153-020-00059-1
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DOI: https://doi.org/10.1007/s43153-020-00059-1