Immobilization of Moniliella spathulata R25L270 Lipase on Ionic, Hydrophobic and Covalent Supports: Functional Properties and Hydrolysis of Sardine Oil

Springer Science and Business Media LLC - Tập 22 Số 10 - Trang 1508
Lívia Souza1, Sonia Moreno‐Pérez2, Gloria Fernández Lorente3, Eliane Pereira Cipolatti4, Débora de Olíveira4, Rodrigo R. Resende1,5, Benevides C. Pessela3,6
1Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Caixa Postal 486, Belo Horizonte MG 31270-901, Brazil
2Pharmacy and Biotechnology Department, School of Biomedical Sciences, Universidad Europea, Villaviciosa de Odón, 28670 Madrid, Spain
3Departamento de Biotecnología y Microbiología de Alimentos, Instituto de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), Campus de la Universidad Autónoma de Madrid, Nicolás Cabrera 9, 28049 Madrid, Spain
4Departamento de Engenharia Química e Engenharia de Alimentos, Universidade Federal de Santa Catarina (UFSC), P.O. Box 476, Florianópolis SC 88040-900, Brazil
5Instituto Nanocell, Divinópolis MG 35500-041, Brazil
6Departamento de Engenharia e Tecnologías, Instituto Superior Politécnico de Tecnologías e Ciências (ISPTEC) Av. Luanda Sul, Rua Lateral Via S10, P.O. Box 1316, Talatona-Luanda Sul, Angola

Tóm tắt

The oleaginous yeast Moniliella spathulata R25L270 was the first yeast able to grow and produce extracellular lipase using Macaúba (Acrocomia aculeate) cake as substrate. The novel lipase was recently identified, and presented promising features for biotechnological applications. The M. spathulata R25L270 lipase efficiently hydrolyzed vegetable and animal oils, and showed selectivity for generating cis-5,8,11,15,17-eicosapentaenoic acid from sardine oil. The enzyme can act in a wide range of temperatures (25–48 °C) and pH (6.5–8.4). The present study deals with the immobilization of M. spathulata R25L270 lipase on hydrophobic, covalent and ionic supports to select the most active biocatalyst capable to obtain omega-3 fatty acids (PUFA) from sardine oil. Nine immobilized agarose derivatives were prepared and biochemically characterized for thermostability, pH stability and catalytic properties (KM and Vmax). Ionic supports improved the enzyme–substrate affinity; however, it was not an effective strategy to increase the M. spathulata R25L270 lipase stability against pH and temperature. Covalent support resulted in a biocatalyst with decreased activity, but high thermostability. The enzyme was most stabilized when immobilized on hydrophobic supports, especially Octyl-Sepharose. Compared with the free enzyme, the half-life of the Octyl-Sepharose derivative at 60 °C increased 10-fold, and lipase stability under acidic conditions was achieved. The Octyl-Sepharose derivative was selected to obtain omega-3 fatty acids from sardine oil, and the maximal enzyme selectivity was achieved at pH 5.0.

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Tài liệu tham khảo

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Springer Science and Business Media LLC

Tập 22 Số 10

1508

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