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Issue 32, 2019
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Biocatalytic transamination in a monolithic flow reactor: improving enzyme grafting for enhanced performance

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Abstract

Transaminases were immobilized onto macrocellular silica monoliths and used for carrying a continuous flow mode transamination reaction. Monoliths were prepared via an emulsion-templated sol–gel method and functionalised by amino-moieties (3-aminopropyl-triethoxysilane, APTES) in order to covalently immobilize the enzymes, using glutaraldehyde as a cross-linking agent. In order to obtain higher performance and improved reproducibility, we investigate the key parameters of APTES functionalisation and of enzyme grafting. Four functionalisation protocols were studied. We show that enhancing the homogeneity of the APTES grafting and controlling the moisture level during functionalisation led to a 3-fold increase in activity as compared to the previously reported data, and greatly improved the reproducibility. Additionally, we report a strong beneficial effect of running the enzyme immobilisation at room temperature instead of 4 °C, further enhancing the obtained activity. Finally, the popular method which consists of stabilizing the covalent attachment of the enzyme by reducing the imine bonds formed between the enzyme and the functionalised surface was investigated. We highlight a strong enzyme deactivation caused by cyanoborohydride, making this strategy irrelevant in this case. The improvements presented here led to more active macrocellular monoliths, of general interest for continuous flow mode biocatalysis.

Graphical abstract: Biocatalytic transamination in a monolithic flow reactor: improving enzyme grafting for enhanced performance

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Publication details

The article was received on 31 Mar 2019, accepted on 05 Jun 2019 and first published on 12 Jun 2019


Article type: Paper
DOI: 10.1039/C9RA02433F
RSC Adv., 2019,9, 18538-18546
  • Open access: Creative Commons BY license
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    Biocatalytic transamination in a monolithic flow reactor: improving enzyme grafting for enhanced performance

    L. van den Biggelaar, P. Soumillion and D. P. Debecker, RSC Adv., 2019, 9, 18538
    DOI: 10.1039/C9RA02433F

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