Title: Broadband laser-based mid-IR spectroscopy for analysis of proteins and monitoring of enzyme activity
Authors: Schwaighofer, Andreas  
Akhgar, Christopher Karim 
Lendl, Bernhard  
Category: Original Research Article
Issue Date: 15-May-2021
Journal: Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 
ISSN: 1386-1425
Laser-based infrared (IR) spectroscopy is an emerging key technology for the analysis of solutes and for real-time reaction monitoring in liquids. Larger applicable pathlengths compared to the traditional gold standard Fourier transform IR (FTIR) spectroscopy enable robust measurements of analytes in a strongly absorbing matrix such as water. Recent advancements in laser development also provide large accessible spectral coverage thus overcoming an inherent drawback of laser-based IR spectroscopy. In this work, we benchmark a commercial room temperature operated broadband external cavityquantum cascade laser (EC-QCL)-IR spectrometer with a spectral coverage of 400 cm 1 against FTIR spectroscopy and showcase its application for measuring the secondary structure of proteins in water, and for monitoring the lipase-catalyzed saponification of triacetin. Regarding the obtained limit of detection (LOD), the laser-based spectrometer compared well to a research-grade FTIR spectrometer employing a liquid nitrogen cooled detector. With respect to a routine FTIR spectrometer equipped with a room temperature operated pyroelectric detector, a 15-fold increase in LOD was obtained in the spectral range of 1600–1700 cm 1. Characteristic spectral features in the amide I and amide II region of three representative proteins with different secondary structures could be measured at concentrations as low as 0.25 mg mL 1. Enzymatic hydrolysis of triacetin by lipase was monitored, demonstrating the advantage of a broad spectral coverage for following complex chemical reactions. The obtained results in combination with the portability and small footprint of the employed spectrometer opens a wide range of future applications in protein analysis and industrial process control, which cannot be readily met by FTIR spectroscopy without recurring to liquid nitrogen cooled detectors.
Keywords: Mid-infrared; Quantum Cascade Laser; Protein secondary structure; Enzyme activity monitoring
DOI: 10.1016/j.saa.2021.119563
Organisation: E164-02-1 - Forschungsgruppe Prozessanalytik 
License: CC BY 4.0 CC BY 4.0
Publication Type: Article
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