Application of infrared spectroscopy (attenuated total reflection) for monitoring enzymatic activity on substrate films

Journal of Biotechnology
Torben Snabe, Steffen Bjørn Petersen

Abstract

Infrared film analysis, a method based on infrared spectroscopy in the mode of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), is demonstrated as a novel analytical method for monitoring enzymatic activity on surface-attached substrate films in the mid infrared range (400-4000 cm(-1)). The ATR-FTIR technique is sensitive to molecules within a distance of approximately 1 microm from the ATR sampling unit surface (a 7 cm(2) hydrophobic ZnSe crystal). Applying a 0.2-0.3 microm thick film on the ATR unit surface, any chemical changes within this film as well as at the interface can be continuously monitored, even having an aqueous phase on top of the film. Infrared film analysis is considered especially useful for studying detergent enzymes, which act on surface bound films consisting of food component like vegetable oils (triacylglycerols) and carbohydrates (e.g. starch). Experimental data are presented for hydrolysis of a triacylglycerol film (triolein) by use of a triacylglycerol lipase (cutinase), and starch film degradation by use of an alpha-amylase.

References

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Citations

Nov 20, 2003·Chemistry and Physics of Lipids·Torben Snabe, Steffen Bjørn Petersen
Sep 25, 2015·The Plant Journal : for Cell and Molecular Biology·Frederick J WarrenPeter R Ellis
Jul 14, 2012·Biotechnology Journal·Antonino NatalelloFrancesco Secundo
Dec 14, 2004·Chemistry and Physics of Lipids·Torben SnabeSteffen Bjørn Petersen
Apr 14, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Kenan P FearsHyun Koo
Aug 24, 2005·Biochemistry·Kristian R PoulsenSteffen B Petersen

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