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Effect of the Presence of Surfactants and Immobilization Conditions on Catalysts’ Properties of Rhizomucor miehei Lipase onto Chitosan

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Abstract

Lipase from Rhizomucor miehei (RML) was immobilized onto chitosan support in the presence of some surfactants added at low levels using two different strategies. In the first approach, the enzyme was immobilized in the presence of surfactants on chitosan supports previously functionalized with glutaraldehyde. In the second one, after prior enzyme adsorption on chitosan beads in the presence of surfactants, the complex chitosan beads-enzyme was then cross-linked with glutaraldehyde. The effects of surfactant concentrations on the activities of free and immobilized RML were evaluated. Hexadecyltrimethylammonium bromide (CTAB) promoted an inhibition of enzyme activity while the nonionic surfactant Triton X-100 caused a slight increase in the catalytic activity of the free enzyme and the derivatives produced in both methods of immobilization. The best derivatives were achieved when the lipase was firstly adsorbed on chitosan beads at 4 °C for 1 h, 220 rpm followed by cross-link the complex chitosan beads-enzyme with glutaraldehyde 0.6% v.v−1 at pH 7. The derivatives obtained under these conditions showed high catalytic activity and excellent thermal stability at 60° and 37 °C. The best derivative was also evaluated in the synthesis of two flavor esters namely methyl and ethyl butyrate. At non-optimized conditions, the maximum conversion yield for methyl butyrate was 89%, and for ethyl butyrate, the esterification yield was 92%. The results for both esterifications were similar to those obtained when the commercial enzyme Lipozyme® and free enzyme were used in the same reaction conditions and higher than the one achieved in the absence of the selected surfactant.

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Acknowledgements

The authors would like to thank the Brazilian research-funding agencies FUNCAP, CNPq, and CAPES.

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  1. Departamento de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, Bloco 709, Fortaleza, CE, CEP 60455-760, Brazil

    Ulisses M. F. de Oliveira, Leonardo J. B. Lima de Matos, Bruna B. Pinheiro & Luciana R. B. Gonçalves

  2. Instituto de Engenharias e Desenvolvimento Sustentável, Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Acarape, CE, CEP 62785-000, Brazil

    Maria Cristiane M. de Souza & José C. S. dos Santos

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  1. Ulisses M. F. de Oliveira
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Correspondence to José C. S. dos Santos or Luciana R. B. Gonçalves.

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de Oliveira, U.M.F., Lima de Matos, L.J.B., de Souza, M.C.M. et al. Effect of the Presence of Surfactants and Immobilization Conditions on Catalysts’ Properties of Rhizomucor miehei Lipase onto Chitosan. Appl Biochem Biotechnol 184, 1263–1285 (2018). https://doi.org/10.1007/s12010-017-2622-1

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