Issue 17, 2023

FTIR microscopy for direct observation of conformational changes on immobilized ω-transaminase: effect of water activity and organic solvent on biocatalyst performance

Abstract

Enzyme immobilization is a key strategy to expand the scope of enzyme applications and to enable the recycling of biocatalysts, resulting in greener and more cost-efficient processes. The full exploitation of the technology advantages is strictly connected to the optimal selection of the carriers and the rational development of the immobilization protocol. The present study achieved such objectives by investigating the activity of a ω-transaminase in organic solvent (toluene) upon immobilization on commercially controlled porosity glass carriers (EziG™) with diverse porosity and surface functionalization. In addition to more conventional wet-chemistry approaches and confocal microscopy, infrared microspectroscopy and imaging were exploited to highlight the enzyme distribution in a label-free manner and provide details on the immobilized enzyme's conformation with respect to the native form. Contrary to what could be expected, the highest activity of the enzyme in organic solvent was achieved for the immobilization protocol on the most hydrophilic support that more severely affects the enzyme secondary structure, promoting a beta-sheet rich folding. Experimental data show that values of water activity above 0.90 in the reaction system had a positive effect on the efficiency of the transaminase reaction. The present study represents the first example of rational development of immobilization protocols relying on direct observation of the enzyme conformation upon immobilization, shedding light on the mutual interaction between the diverse process parameters and the carrier properties.

Graphical abstract: FTIR microscopy for direct observation of conformational changes on immobilized ω-transaminase: effect of water activity and organic solvent on biocatalyst performance

Supplementary files

Article information

Article type
Paper
Submitted
13 Nov. 2022
Accepted
23 Maijs 2023
First published
22 Jūn. 2023
This article is Open Access
Creative Commons BY-NC license

Catal. Sci. Technol., 2023,13, 4955-4967

FTIR microscopy for direct observation of conformational changes on immobilized ω-transaminase: effect of water activity and organic solvent on biocatalyst performance

I. Rădoi, D. E. Bedolla, L. Vaccari, A. Todea, F. Zappaterra, A. Volkov and L. Gardossi, Catal. Sci. Technol., 2023, 13, 4955 DOI: 10.1039/D2CY01949C

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