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PublicationBiotechnology Advances
Pages 746-770

Immobilization of lipases on hydrophobic supports: immobilization mechanism, advantages, problems, and solutions

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimization of biocatalysts and bioprocesses
Rafael C. Rodrigues1, Jose J. Virgen-Ortíz2, José C.S. dos Santos3, Ángel Berenguer-Murcia4, Andres R. Alcantara5, Oveimar Barbosa6, Claudia Ortiz7, Roberto Fernandez-Lafuente8,*
1Biocatalysis and Enzyme Technology Lab, Institute of Food Science and Technology, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, 9500, P.O. Box 15090, Porto Alegre, RS, Brazil.
2CONACYT - Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD)  - CIDAM, Km. 8 Antigua Carretera a Pátzcuaro s/n, 58341 Morelia, Michoacán, México.
3Instituto de Engenharias e Desenvolvimento Sustentável, Universidade da Integração Internacional da Lusofonia Afro-Brasileira, CEP 62790-970, Redenção, CE, Brazil.
4 Departamento de Química Inorgánica e Instituto Universitario de Materiales, Universidad de Alicante, Campus de San Vicente del Raspeig, Ap. 99 - 03080 Alicante, Spain.
5Departmento de Química en Ciencias Farmacéuticas,  Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal, s/n. 28040-Madrid, Spain
6Departamento de Química, Facultad de Ciencias. Universidad del Tolima, Ibagué, Colombia.
7Escuela de Microbiología, Universidad Industrial de Santander, Bucaramanga, Colombia

8 Departamento de Biocatálisis ICP-CSIC. Campus UAM-CSIC. Cantoblanco. 28049 Madrid. Spain.

Lipases are the most widely used enzymes in biocatalysis, and the most utilized method for enzyme immobilization is using hydrophobic supports at low ionic strength. This method allows the one step immobilization, purification, stabilization, and hyperactivation of lipases, and that is the main cause of their popularity. This review focuses on these lipase immobilization supports. First, the advantages of these supports for lipase immobilization will be presented and the likeliest immobilization mechanism (interfacial activation on the support surface) will be revised. Then, its main shortcoming will be discussed: enzyme desorption under certain conditions (such as high temperature, presence of cosolvents or detergent molecules). Methods to overcome this problem include physical or chemical crosslinking of the immobilized enzyme molecules or using heterofunctional supports. Thus, supports containing hydrophobic acyl chain plus epoxy, glutaraldehyde, ionic, vinylsulfone or glyoxyl groups have been designed. This prevents enzyme desorption and improved enzyme stability, but it may have some limitations, that will be discussed and some additional solutions will be proposed (e.g., chemical amination of the enzyme to have a full covalent enzyme-support reaction). These immobilized lipases may be subject to unfolding and refolding strategies to reactivate inactivated enzymes. Finally, these biocatalysts have been used in new strategies for enzyme coimmobilization, where the most stable enzyme could be reutilized after desorption of the least stable one after its inactivation.

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