Real‐Time Monitoring of Enzyme‐Catalysed Reactions using Deep UV Resonance Raman Spectroscopy



Westley, C, Fisk, H, Xu, Y ORCID: 0000-0003-3228-5111, Hollywood, KA, Carnell, AJ ORCID: 0000-0002-6353-403X, Micklefield, J, Turner, NJ and Goodacre, R ORCID: 0000-0003-2230-645X
(2017) Real‐Time Monitoring of Enzyme‐Catalysed Reactions using Deep UV Resonance Raman Spectroscopy. Chemistry: A European Journal, 23 (29). 6983 - 6987.

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Abstract

For enzyme‐catalysed biotransformations, continuous in situ detection methods minimise the need for sample manipulation, ultimately leading to more accurate real‐time kinetic determinations of substrate(s) and product(s). We have established for the first time an on‐line, real‐time quantitative approach to monitor simultaneously multiple biotransformations based on UV resonance Raman (UVRR) spectroscopy. To exemplify the generality and versatility of this approach, multiple substrates and enzyme systems were used involving nitrile hydratase (NHase) and xanthine oxidase (XO), both of which are of industrial and biological significance, and incorporate multistep enzymatic conversions. Multivariate data analysis of the UVRR spectra, involving multivariate curve resolution‐alternating least squares (MCR‐ALS), was employed to effect absolute quantification of substrate(s) and product(s); repeated benchmarking of UVRR combined with MCR‐ALS by HPLC confirmed excellent reproducibility.

Item Type: Article
Uncontrolled Keywords: biotransformations, quantification, on-line monitoring, reaction monitoring, UV resonance Raman (UVRR) spectroscopy
Depositing User: Symplectic Admin
Date Deposited: 30 Mar 2017 13:43
Last Modified: 01 Sep 2021 07:12
DOI: 10.1002/chem.201701388
Open Access URL: https://chemistry-europe.onlinelibrary.wiley.com/d...
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URI: https://livrepository.liverpool.ac.uk/id/eprint/3006725