Real-time monitoring of enzyme-catalysed reactions using deep UV resonance Raman spectroscopy

Research output: Contribution to journalArticle

  • External authors:
  • Chloe Westley
  • Heidi Fisk
  • Yunji Xu
  • Katherine Hollywood
  • Andrew J. Carnell

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.

Bibliographical metadata

Original languageEnglish
Pages (from-to)6983-6987
Number of pages5
JournalChemistry: A European Journal
Volume23
Issue number29
Early online date2 May 2017
DOIs
StatePublished - 23 May 2017