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Covalent Immobilization of Chondrostereum purpureum Endopolygalacturonase on Ferromagnetic Nanoparticles: Catalytic Properties and Biotechnological Application

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Abstract

Pectinases are widely used in a variety of industrial processes. However, their application is limited by low catalytic processivity, reduced stability, high cost, and poor re-use compatibility. These drawbacks may be overcome by enzyme immobilization with ferromagnetic nanoparticles, which are easily recovered by a magnetic field. In this work, an endopolygalacturonase from Chondrostereum purpureum (EndoPGCp) expressed in Pichia pastoris was immobilized on glutaraldehyde-activated chitosan ferromagnetic nanoparticles (EndoPGCp-MNP) and used to supplement a commercial enzyme cocktail. No significant differences in biochemical and kinetic properties were observed between EndoPGCp-MNP and EndoPGCp, although the EndoPGCp-MNP showed slightly increased thermostability. Cocktail supplementation with EndoPGCp-MNP increased reducing sugar release from orange wastes by 1.8-fold and showed a synergistic effect as compared to the free enzyme. Furthermore, EndoPGCp-MNP retained 65% of the initial activity after 7 cycles of re-use. These properties suggest that EndoPGCp-MNP may find applications in the processing of pectin-rich agroindustrial residues.

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Acknowledgements

We thank André Justino for the technical assistance.

Funding

This investigation was supported by research grants from CNPq (Conselho de Desenvolvimento Científico e Tecnológico), FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo), and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Finance Code 001). S.C. received a Ph.D. scholarship from FAPESP (2017/13734–3); L.P.M. received post-doctoral scholarship from FAPESP (2016/17582–0 and 2019/17958–9); J.C.S.S. received post-doctoral scholarship from FAPESP (2019/21989–7); and R.J.W. received researcher stipend from CNPq 305788/2017–5 and the National Institute of Science and Technology of Bioetanol (INCT-Bioetanol) (FAPESP 2011/57908–6 and 2014/50884–5, CNPq 574002/2008–1, and 465319/2014–9).

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  1. Departamento de Química, Faculdade de Filosofia, Ciências E Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, 14040-901, Brazil

    Sibeli Carli, Jose Carlos Santos Salgado, Luana Parras Meleiro & Richard John Ward

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  2. Jose Carlos Santos Salgado
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Sibeli Carli: data curation, formal analysis, investigation, methodology, figure preparation, visualization, writing—review & editing; Jose Carlos Santos Salgado: data curation, formal analysis, investigation, methodology, figure preparation, visualization, writing—review & editing; Luana Parras Meleiro: data curation, formal analysis, investigation, methodology, figure preparation, visualization, writing—original draft; Richard John Ward: conceptualization, funding acquisition, project administration, resource management, team supervision, writing—review & editing.

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Carli, S., Salgado, J.C.S., Meleiro, L.P. et al. Covalent Immobilization of Chondrostereum purpureum Endopolygalacturonase on Ferromagnetic Nanoparticles: Catalytic Properties and Biotechnological Application. Appl Biochem Biotechnol 194, 848–861 (2022). https://doi.org/10.1007/s12010-021-03688-5

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