Immobilisation of Candida rugosa lipase on polyhydroxybutyrate via a combination of adsorption and cross-linking agents to enhance acylglycerol production |
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| Institution: | 1. Biocatalysis 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;2. CONACYT - Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD) - CIDAM, Km. 8 Antigua Carretera a Pátzcuaro s/n, 58341 Morelia, Michoacán, México;3. Instituto 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;5. Departmento de Química en Ciencias Farmacéuticas, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal, s/n, 28040 Madrid, Spain;6. Departamento de Química, Facultad de Ciencias, Universidad del Tolima, Ibagué, Colombia;7. Escuela de Microbiología, Universidad Industrial de Santander, Bucaramanga, Colombia;8. Departamento de Biocatálisis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain |
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| Abstract: | In the present study, a combination of immobilisation processes was utilised to prepare robust biocatalysts. First, lipase from Candida rugosa was adsorbed on polyhydroxybutyrate (PHB) particles, followed by cross-linking with glutaraldehyde. Conditions for creating immobilised lipase involved the addition of 0.6 M glutaraldehyde and 45 U mL?1 lipase while mixing at 150 rpm (4 °C) for 30 min. These conditions produced the highest yield of immobilised lipase (92 %) and the highest levels of activity (1.94 mg g?1 support). At 40 °C and pH 9 the immobilised enzyme was optimally active with a Km and Vmaxat 1.2 mM and 2.5 × 10-3 mmol min?1, respectively. The use of immobilised lipase improved thermal stability, storage stability, and reusability.The immobilised lipase retained 80 % of its activity after incubation at 30–60 °C for 2 h and 4 °C for 30 d in 0.2 M sodium phosphate buffer (pH 7.0). Moreover, the immobilised enzyme retained 50 % of its activity after more than 14 cycles under optimal conditions. The immobilised lipase was used to produce monoacylglycerol MAG. The existence of a carbonyl group at 1,743 and 1,744 cm?1 was identified using attenuated total reflectance (ATR)-Fourier transformed infrared spectroscopy. Results showed that 48 % MAG was produced. |
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| Keywords: | Immobilised enzyme Glycerolysis Lipase Monoacylglycerol Polyhydroxyalkanoate |
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