title: The effect of cultivation media and washing whole-cell biocatalysts on monoamine oxidase catalyzed oxidative desymmetrization of 3-azabicyclo[3,3,0]octane.
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-azabicyclo[3,3,0]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.
authors.affiliation: Department of Chemical and Biochemical Engineering, Building 229, Søltofts Plads, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
authors.forename: Hemalata
authors.initials: H
authors.lastname: Ramesh
authors.affiliation: Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37 Bratislava, Slovakia.
authors.forename: Petra
authors.initials: P
authors.lastname: Zajkoska
authors.affiliation: Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37 Bratislava, Slovakia.
authors.forename: Martin
authors.initials: M
authors.lastname: Rebroš
authors.affiliation: Department of Chemical and Biochemical Engineering, Building 229, Søltofts Plads, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. Electronic address: jw@kt.dtu.dk.
authors.forename: John M
authors.initials: JM
authors.lastname: Woodley
mesh_terms: D001372:Aza Compounds / Q000737:chemistry* / Q000378:metabolism*; D053961:Azabicyclo Compounds / Q000737:chemistry* / Q000378:metabolism*; D055162:Biocatalysis; D003470:Culture Media; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism; D005285:Fermentation; D007218:Industrial Microbiology; D007700:Kinetics; D008995:Monoamine Oxidase / Q000378:metabolism*; D009795:Octanes / Q000737:chemistry* / Q000378:metabolism*; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D001372:Aza Compounds; D053961:Azabicyclo Compounds; D003470:Culture Media; D009795:Octanes; D011994:Recombinant Proteins; C518626:azabicyclo(3.3.0)-octane; D008995:Monoamine Oxidase
keywords: Monoamine oxidase; Resting state; Washing; Whole-cell biocatalysis