The effect of cultivation media and washing whole-cell biocatalysts on monoamine oxidase catalyzed oxidative desymmetrization of 3-azabicyclo[3,3,0]octane |
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Affiliation: | 1. Department of Chemical and Biochemical Engineering, Building 229, Søltofts Plads, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark;2. Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37 Bratislava, Slovakia;1. Department of Medicinal Chemistry, Collegium Medicum in Bydgoszcz, Faculty of Pharmacy, Nicolaus Copernicus University in Toruń, Dr. A. Jurasza 2, 85-089 Bydgoszcz, Poland;2. Chair of Chemistry and Photochemistry of Polymers, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, Gagarina 7, 87-100 Toruń, Poland;1. Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal;2. Centre for Biomedical Research (CBMR), University of Algarve, Faro, Portugal;1. The Ohio State Biochemistry Program, Columbus, OH 43210, USA;2. Department of Chemistry and Biochemistry, Columbus, OH 43210, USA;3. The Ohio State University Biophysics Program, The Ohio State University, Columbus, OH 43210, USA;4. Department of Chemistry, University of Connecticut, Storrs, CT 06269, USA;1. Institute of Biomedical Chemistry, Pogodinskaya st 10/8, 119121 Moscow, Russia;2. Peoples Friendship University of Russia (RUDN University), Miklukho-Maklaya 6, 117198 Moscow, Russia;3. N.N. Blokhin Cancer Research Center, Kashirskoe Shosse 24, 115478 Moscow, Russia |
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Abstract: | It is well known that washing whole-cells containing enzyme activities after fermentation, but prior to biocatalysis can improve their activity in the subsequent reaction. In this paper, we quantify the impact of both the fermentation media and cell washing on the performance of whole-cell biocatalysis. The results are illustrated using a recombinant monoamine oxidase (expressed in Escherichia coli, used in resting state) for the oxidative desymmetrization of 3-azabicyclo3,3,0]octane. It was shown that the need for washing biocatalyst prior to use in a reaction is dependent upon growth medium. Unlike cells grown in LB medium, washing of the cells was essential for cells grown on TB medium. With TB media, washing the cells improved the final conversion by approximately a factor of two. Additionally, over 50-fold improvement was achieved in initial activity. A potential reason for this improvement in activity was identified to be the increase in transfer of substrates across the cell membrane as a result of cell washing. |
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Keywords: | Whole-cell biocatalysis Washing Resting state Monoamine oxidase |
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