STERILIZABLE MINIATURE BIOREACTOR PLATFORM FOR ANAEROBIC FERMENTATION PROCESS
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Published:
Feb 28, 2021
Keywords:
Miniature bioreactor anaerobic fermentation scale down miniaturization S.cerevisiae
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Abstract
This paper presents the establishment of a miniature bioreactor platform for anaerobic microbial fermentation processes. It is made from a universal glass bottle and has a working volume of 16 mL. Reactor features included mixing via magnetic stirrer, temperature control via electrical heater and cells optical density (OD) sensing. All sensors and actuators integrated into the reactor were operated using LabVIEWTM (National Instrument, TX, US). The top lid of the bottle was modified to include a 3mm poly(methylmethacrylate) (PMMA) polymer layer where it was machined to provide spaces for fluidic ports and integration of sensors. Each reactor components were sterilized prior to conducting fermentation experiments. Main body of the reactor was made of glass and it was sterilized using the standard heat sterilization method (121oC for 15 minutes) where else other components were sterilized by exposure to UV light for 2 hours. A series of anaerobic fermentation experiments were conducted in batch mode using S.cerevisiae to evaluate the workability of the system. Fermentation experiments were conducted using inoculum concentration of 2 g∙L-1 and starting glucose concentration between 10 g∙L-1 and 20 g∙L-1. In every experiment, mixing was set to operate at 400 rpm and temperature was adjusted to 30 ± 2oC. Experiments were carried out until stationary phase was attained. Under these conditions, the best fermentation profile was obtained with glucose concentration of 20 /L where cell specific growth rate was found to be about 0.28 h-1. Bench marking step was also performed where results attained in a miniature bioreactor platform were comparable with the one attained using a 50 mL flask.
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Arriafdi, Z., Ramachendrin, A., & Zainal Alam, M. N. H. (2021). STERILIZABLE MINIATURE BIOREACTOR PLATFORM FOR ANAEROBIC FERMENTATION PROCESS. Malaysian Journal of Science, 40(1), 22–33. https://doi.org/10.22452/https://doi.org/10.22452/mjs.vol40no1.2
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Zainal Alam, M., Schäpper, D. and Gernaey, K. (2010). Embedded resistance wire as a heating element for temperature control in microbioreactors. Journal of Micromechanics and Microengineering, 20(5): 055014.
Zainal Alam, M. N. H., Schäpper, D., and Gernaey, K. V. (2012). ‘Establishment of a Gaseous pH concept in microbioreactors’. International Conference on Food Engineering & Biotechnology (3rd ICFEB), Sepang, Malaysia. ISSN: 2010-4618. Vol3: pg 228-234.
Zelenev, V.V., van Bruggen, A.H.C., and Semenov, A.M. (2005). Modeling wave-like dynamics of oligotrophic and copiotrophic bacteria along wheat roots in response to nutrient input from a growing root tip. Ecological Modelling. 188, 404 – 417.
Bolic, A. (2018). Development and application of a milliliter-scale bioreactor for continuous microbial
cultivations. Thesis of Doctoral Degree, Denmark Technical University.
Bolic, A., Larsson, H., Hugelier, S., Lantz, A.E., Krühne, U., and Gernaey, K.V. (2016). A flexible well-mixed milliliter-scale reactor with high oxygen transfer rate for microbial cultivations. Chemical Engineering Journal. 303, 655–666.
Halimoon, H., Hussain, A., Kouzani, A., & Alam, M. (2016). Aerobic fermentation of Saccharomeyes cerevisae in a miniature bioreactor made of low cost poly(methylmethacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS) polymers. Sains Malaysiana, 45(6), 969-976.
Kheradmandnia, S., Hashemi-Najafabadi, S., Shojaosadati, S., Mousavi, S., and Malek Khosravi, K. (2014). Development of parallel miniature bubble column bioreactors for fermentation process. Journal of Chemical Technology & Biotechnology, 90(6), 1051-1061.
Kirk, T. and Szita, N. (2013). Oxygen transfer characteristics of miniaturized bioreactor systems. Biotechnology and Bioengineering, 110(4), 1005-1019.
Krakau, K. (2016). Standard Operating Procedures for a Single-Use Fermenter. Thesis of Bachelor Degree, Technical University of Denmark.
Najafpour, G. (2007). Bioreactor Design. Biochemical Engineering and Biotechnology, pp.142-169.
Panatto, D., Amicizia, D., Bragazzi, N., Rizzitelli, E., Tramalloni, D., Valle, I. and Gasparini,R. (2015) ‘Human Papillomavirus Vaccine. Advances in
Protein Chemistry and Structural Biology’, 231-322.
Schäpper, D., Zainal Alam, M.N.H., Szita, N., Lantz, A.E. & Gernaey, K.V. 2009. Application of microbioreactors in fermentation process development: A review. Analytical and Bioanalytical Chemistry 395(3): 679-695.
Schmideder, A., Severin, T., Cremer, J. and Weuster-Botz, D. (2015). A novel milliliter-scale chemostat system for parallel cultivation of microorganisms in stirred-tank bioreactors. Journal of Biotechnology, 210, .19-24.
Zainal Alam, M., Jaya Kumar, J., John Whyte, D., Doeven, E., & Kouzani, A. (2018). A portable sensor for cell optical density measurement in microfluidic chips. Measurement and Control, 51(7-8), 213-222.
Zainal Alam, M., Schäpper, D. and Gernaey, K. (2010). Embedded resistance wire as a heating element for temperature control in microbioreactors. Journal of Micromechanics and Microengineering, 20(5): 055014.
Zainal Alam, M. N. H., Schäpper, D., and Gernaey, K. V. (2012). ‘Establishment of a Gaseous pH concept in microbioreactors’. International Conference on Food Engineering & Biotechnology (3rd ICFEB), Sepang, Malaysia. ISSN: 2010-4618. Vol3: pg 228-234.
Zelenev, V.V., van Bruggen, A.H.C., and Semenov, A.M. (2005). Modeling wave-like dynamics of oligotrophic and copiotrophic bacteria along wheat roots in response to nutrient input from a growing root tip. Ecological Modelling. 188, 404 – 417.