title: Immobilised whole-cell recombinant monoamine oxidase biocatalysis.
abstract: This work demonstrates the first example of the immobilisation of MAO-N whole cells to produce a biocatalyst that remained suitable for repetitive use after 11 months of storage and stable up to 15 months after immobilisation. The production of Escherichia coli expressing recombinant MAO-N was scaled up to bioreactors under regulated, previously optimised conditions (10% DO, pH 7), and the amount of biomass was almost doubled compared to flask cultivation. Subsequently, pilot immobilisation of the whole-cell biocatalyst using LentiKats technology was performed. The amount of the immobilised biomass was optimised and the process was scaled up to a production level by immobilising 15 g of dry cell weight per litre of polyvinyl alcohol to produce 3 kg of whole-cell ready-to-use biocatalyst. The immobilised biocatalyst retained its initial activity over six consecutive biotransformations of the secondary amine model compound 3-azabicylo [3,3,0]octane, a building block of the hepatitis C drug telaprevir. Consecutive cultivation cycles in growth conditions not only increased the initial specific activity of biocatalyst produced on the industrial plant by more than 30%, but also significantly increased the rate of the biotransformation compared to the non-propagated biocatalyst.
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.forename: Michal
authors.initials: M
authors.lastname: Rosenberg
authors.forename: Rachel
authors.initials: R
authors.lastname: Heath
authors.forename: Kirk J
authors.initials: KJ
authors.lastname: Malone
authors.forename: Radek
authors.initials: R
authors.lastname: Stloukal
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Turner
authors.forename: Martin
authors.initials: M
authors.lastname: Rebroš
mesh_terms: D015306:Biogenic Monoamines / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology; D018914:Cells, Immobilized / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008995:Monoamine Oxidase / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015306:Biogenic Monoamines; D011994:Recombinant Proteins; D008995:Monoamine Oxidase