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High-cell density cultivation technique for microplates
S182
Abstracts / Journal of Biotechnology 131S (2007) S133–S187
References Hofvendahl, K., Hahn-H¨agerdal, B., 2000. Factors affecting the fermentative lactic acid production from renewable resources. Enzyme Microb. Technol. 26, 87–107. Bai, D.-M., Jia, M.-Z., Zhao, X.-M., Ban, R., Shen, F., Li, X.-G., Xu, S.-M., 2003. l(+)-Lactic acid production by pellet-form Rhizopus oryzae R1021 in a stirred tank fermentor. Chem. Eng. Sci. 58, 785–791. Mussatto, S.I., Dragone, G., Rocha, G.J.M., Roberto, I.C., 2006. Optimum operating conditions for brewer’s spent grain soda pulping. Carbohydr. Polym. 64, 22–28.
doi:10.1016/j.jbiotec.2007.07.920 91. Controlled high-cell density cultivation in shake flasks Johanna Panula-Per¨al¨a ∗ , Marco Casteleijn, Antti Vasala, Peter Neubauer Bioprocess Engineering Laboratory, University of Oulu, Oulu, Finland Shake flask cultivations are commonly used in laboratories and bio-industries. This method is easy and low cost, but generally the reliability is poor. Because shake flask cultivations are usually performed as batch processes, excess substrate causes fast uncontrolled growth, overflow metabolism and oxygen depletion. Therefore, high cell densities are generally not reached and flask to flask variations occur. In industrial processes, fedbatch method is used to avoid these inhibitive effects and the controlled substrate addition in a fed-batch cultivation ensures that processes are very reproducible. Here we present an “Enzyme based substrate delivery technique” (Enbase) as a new simple solution for a substrate limited fed-batch system in shake flasks. The new technique ensures controlled growth and, consequently higher cell densities are obtained. The growth-limiting substrate is delivered by biocatalytic degradation of a metabolically inactive polymer. In this system the growth rate can be simply controlled by the concentration of the enzyme. With this method, up to four times higher cell densities were reached than in conventional shake flask cultivations. The method is shown to be especially beneficial for recombinant protein production. At the example of a recombinant triosephosphate isomerase we show that not only higher cell densities are obtained in Enbase flasks, but also the level of functional recombinant product is increased compared to reference cultivations. doi:10.1016/j.jbiotec.2007.07.921
92. High-cell density cultivation technique for microplates Johanna Panula-Per¨al¨a 1,∗ , Robert Wilmanowski 1 , Juozas Siurkus 2 , Marco Casteleijn 1 , Antti Vasala 1 , Peter Neubauer 1 1 Bioprocess Engineering Laboratory, University of Oulu, Oulu,
Finland 2 Fermentas Ltd., Vilnius, Lithuania Microplate based cultivation has become an interesting option for parallelisation and automation of small scale cultures. However, often the application is limited by the low cell densities obtained in the small volumes with product yields below the detection limit. A significant limitation are oxygen transfer and pH change. Control strategies as applied in larger scale are not easily applicable in the microscale. Especially the fed-batch technology, which is the typical procedure to obtain high cell densities in larger scale, cannot be applied in the very small scale due to problems with viscosity, delivery of microlitre amounts and mixing. Here we describe a simple new technique, which is based on concerted diffusion and enzyme based substrate delivery for the performance of fed-batch processes at the microlitre scale. The new technology allows to obtain 10-50 times higher cell densities than in standard microscale cultivations. With this technique the growth rate of the culture can be simply controlled by the amount of the enzyme, which releases the substrate from an inactive polymer. Aside from performance parameters of the system we demonstrate here that the new technique can be very beneficial for the production of recombinant proteins at different examples. doi:10.1016/j.jbiotec.2007.07.922 93. Optimization of Lactobacillus casei growth and lactic acid production in batch culture Hassan Korbekandi 1,∗ , Daryoush Abedi 2 , Mohammad Jalali 3 , Mohammad-Reza Fazeli 4 , Marzieh Heidari 2 1 Genetics
& Molecular Biology Department, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 2 Biotechnology Department, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 3 Food Microbiology Department, School of Health, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 4 Biotechnology Department, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran Nowadays, probiotics as natural complements are getting more attention. Lactobacillus casei is one of the most important probiotics, producing l-lactic acid. This optical isomer is used in medical and pharmaceutical industries. In this study we tried to optimize the growth of L. casei and lactic acid production in order to prepare the necessary information for future and scale up studies.
Abstracts / Journal of Biotechnology 131S (2007) S133–S187
References Hofvendahl, K., Hahn-H¨agerdal, B., 2000. Factors affecting the fermentative lactic acid production from renewable resources. Enzyme Microb. Technol. 26, 87–107. Bai, D.-M., Jia, M.-Z., Zhao, X.-M., Ban, R., Shen, F., Li, X.-G., Xu, S.-M., 2003. l(+)-Lactic acid production by pellet-form Rhizopus oryzae R1021 in a stirred tank fermentor. Chem. Eng. Sci. 58, 785–791. Mussatto, S.I., Dragone, G., Rocha, G.J.M., Roberto, I.C., 2006. Optimum operating conditions for brewer’s spent grain soda pulping. Carbohydr. Polym. 64, 22–28.
doi:10.1016/j.jbiotec.2007.07.920 91. Controlled high-cell density cultivation in shake flasks Johanna Panula-Per¨al¨a ∗ , Marco Casteleijn, Antti Vasala, Peter Neubauer Bioprocess Engineering Laboratory, University of Oulu, Oulu, Finland Shake flask cultivations are commonly used in laboratories and bio-industries. This method is easy and low cost, but generally the reliability is poor. Because shake flask cultivations are usually performed as batch processes, excess substrate causes fast uncontrolled growth, overflow metabolism and oxygen depletion. Therefore, high cell densities are generally not reached and flask to flask variations occur. In industrial processes, fedbatch method is used to avoid these inhibitive effects and the controlled substrate addition in a fed-batch cultivation ensures that processes are very reproducible. Here we present an “Enzyme based substrate delivery technique” (Enbase) as a new simple solution for a substrate limited fed-batch system in shake flasks. The new technique ensures controlled growth and, consequently higher cell densities are obtained. The growth-limiting substrate is delivered by biocatalytic degradation of a metabolically inactive polymer. In this system the growth rate can be simply controlled by the concentration of the enzyme. With this method, up to four times higher cell densities were reached than in conventional shake flask cultivations. The method is shown to be especially beneficial for recombinant protein production. At the example of a recombinant triosephosphate isomerase we show that not only higher cell densities are obtained in Enbase flasks, but also the level of functional recombinant product is increased compared to reference cultivations. doi:10.1016/j.jbiotec.2007.07.921
92. High-cell density cultivation technique for microplates Johanna Panula-Per¨al¨a 1,∗ , Robert Wilmanowski 1 , Juozas Siurkus 2 , Marco Casteleijn 1 , Antti Vasala 1 , Peter Neubauer 1 1 Bioprocess Engineering Laboratory, University of Oulu, Oulu,
Finland 2 Fermentas Ltd., Vilnius, Lithuania Microplate based cultivation has become an interesting option for parallelisation and automation of small scale cultures. However, often the application is limited by the low cell densities obtained in the small volumes with product yields below the detection limit. A significant limitation are oxygen transfer and pH change. Control strategies as applied in larger scale are not easily applicable in the microscale. Especially the fed-batch technology, which is the typical procedure to obtain high cell densities in larger scale, cannot be applied in the very small scale due to problems with viscosity, delivery of microlitre amounts and mixing. Here we describe a simple new technique, which is based on concerted diffusion and enzyme based substrate delivery for the performance of fed-batch processes at the microlitre scale. The new technology allows to obtain 10-50 times higher cell densities than in standard microscale cultivations. With this technique the growth rate of the culture can be simply controlled by the amount of the enzyme, which releases the substrate from an inactive polymer. Aside from performance parameters of the system we demonstrate here that the new technique can be very beneficial for the production of recombinant proteins at different examples. doi:10.1016/j.jbiotec.2007.07.922 93. Optimization of Lactobacillus casei growth and lactic acid production in batch culture Hassan Korbekandi 1,∗ , Daryoush Abedi 2 , Mohammad Jalali 3 , Mohammad-Reza Fazeli 4 , Marzieh Heidari 2 1 Genetics
& Molecular Biology Department, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 2 Biotechnology Department, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 3 Food Microbiology Department, School of Health, Isfahan University of Medical Sciences, Isfahan, Islamic Republic of Iran 4 Biotechnology Department, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran Nowadays, probiotics as natural complements are getting more attention. Lactobacillus casei is one of the most important probiotics, producing l-lactic acid. This optical isomer is used in medical and pharmaceutical industries. In this study we tried to optimize the growth of L. casei and lactic acid production in order to prepare the necessary information for future and scale up studies.