Immobilised whole-cell recombinant monoamine oxidase biocatalysis
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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.
KeywordsImmobilisation E. coli MAO-N Biotransformation Amine
The research leading to these results has received funding from the European Union Seventh Framework Programme BIONEXGEN under grant agreement no. 266025. This work was co-funded by the Slovak Research and Development Agency under contract no. DO7RP-0042-11.
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