Agitation,aeration and shear stress as key factors in inulinase production by Kluyveromyces marxianus |
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| Institution: | 1. Laboratorio de Fermentaciones, Centro de Ingeniería Genética y Biotecnología de Sancti Spíritus (CIGBSS), Circunvalante Norte S/N, Olivos 3, Apartado Postal 83, Sancti Spíritus, 60200, Cuba;2. Grupo Tecnología de Enzimas, Dirección de Investigaciones Agropecuarias, Centro de Ingeniería Genética y Biotecnología (CIGB), Ave 31 entre 158 y 190, Apartado Postal 6162, Habana, 10600, Cuba;3. Grupo Virología de Plantas, Dirección de Investigaciones Agropecuarias, Centro de Ingeniería Genética y Biotecnología (CIGB), Ave 31 entre 158 y 190, Apartado Postal 6162, Habana, 10600, Cuba;1. Graduation in Civil and Environmental Engineering, University of Passo Fundo (UPF), Campus I, km 171, BR 285, P.O. Box 611, 99001-970 Passo Fundo, Rio Grande do Sul, Brazil;2. Food Engineering Course, University of Passo Fundo (UPF), Campus I, km 171, BR 285, P.O. Box 611, 99001-970 Passo Fundo, Rio Grande do Sul, Brazil;3. Graduation in Environmental Science and Technology, Federal University of Fronteira Sul – Campus Erechim, RS 135, Km 72, 99700-000 Erechim, RS, Brazil;4. Graduation in Food Technology, Federal Technological University of Paraná (UTFPR), Av. Brasil, 4232, 85884-000, P.O. Box 271, Medianeira, Paraná, Brazil;1. Centre for Biological Engineering, Loughborough University, Leicestershire LE11 3TU, UK;2. School of Chemical Engineering, University of Birmingham, Birmingham B15 2TT, UK;3. Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK;4. Department of Bioengineering, Instituto Superior Técnico (IST), Universidade de Lisboa, Portugal;1. Tecnológico Nacional de México, Instituto Tecnológico de Durango, Departamento de Ingenierías Química y Bioquímica, Felipe Pescador 1803 Ote, Colonia Nueva Vizcaya, Durango, Dgo C.P. 34080, Mexico;2. Facultad de Ingeniería en Tecnología de la Madera (FITECMA), Edificio D, CU, Universidad Michoacana de San Nicolás de Hidalgo (UMSNH), Av. Fco. J. Mújica S/N. Col Felicitas de Río, Morelia, Michoacán C.P. 58040, Mexico;3. Centro de Investigación en Materiales Avanzados S. C. (CIMAV-Unidad Monterrey), Grupo de Materiales Funcionales y Nanotecnología, Alianza Norte 202, Autopista Monterrey-Aeropuerto km 10, Apodaca, Nuevo León C.P. 66628, Mexico;4. Departamento de Investigación de Alimentos, Facultad de Química, Universidad Autónoma de Coahuila, Blvd. V. Carranza e Ing. José Cárdenas V. S/N. Col. República Ote, Saltillo, Coahuila C.P. 25280, Mexico;5. Departamento de Biotecnología, Universidad Autónoma Metropolitana, PA 55-535, Iztapalapa, C.P. 09340 CDMX, Mexico |
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| Abstract: | Factorial design and response surface analyses were used to optimize the production of inulinase (2,1-β-d-fructan fructanohydrolase, EC 3.2.1.7) by Kluyveromyces marxianus ATCC 16045, using sucrose as carbon source. Effects of aeration, agitation and type of impeller (disk turbine, marine, pitched blade) were studied in a batch stirred reactor. Two factorial designs 22 were carried out. Agitation speed varied from 50 to 550 rpm (revolution per minute), aeration rate from 0.5 to 2.0 vvm (air volume/broth volume·minute). It has been shown that the enzyme production was strongly influenced by mixing conditions, while aeration rate was shown to be less significant. Additionally, the increase in the agitation speed is limited by the death rate, which increases drastically at high speeds, lowering the enzyme production. Also, the impeller type has significant influence in the production, the disk impeller at 450 rpm and aeration at 1.0 vvm led to an activity of 121 UI/mL, while the pitched blade was shown to be the best impeller for this process, leading to the best production, 176 UI/mL, at 450 rpm and 1.0 vvm. The maximum shear stress for inulinase production was about 0.22 Pa, since higher values cause higher cell death rates, affecting the enzyme production. The same results were confirmed with another microorganism, which was also sensible to shear stress. Therefore, it has been concluded that in some cases, mainly when the microorganism is sensible to shear stress, the interaction between mass transfer and mechanical stress should be considered in scale up processes. |
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