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In Situ Tracking of Enzymatic Breakdown of Starch Granules by Synchrotron UV Fluorescence Microscopy

Abstract : Synchrotron UV fluorescence microscopy was used for the first time to visualize the adsorption and diffusion of an enzyme while degrading a solid substrate. The degradation pathway of single starch granules by two amylases, optimized for biofuel production and industrial starch hydrolysis, was followed by tryptophan fluorescence (excitation at 280 nm, emission filter at 300-400 nm) and visible light imaging. Thus, both the adsorption of enzyme onto starch granules at 283 nm resolution and the resulting morphological changes,were recorded at different stages of hydrolysis. It is the first time that amylases were localized on starch without staining or adding a fluorescent probe at such high resolution. This technique presents a very high potential for imaging proteins in complex systems. Its sensitivity was demonstrated by the detection of GBSS (the granular bound starch synthase) at high recording times, GBSS being present at very low levels in maize starch granules.
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https://hal.inrae.fr/hal-02649349
Contributor : Migration Prodinra <>
Submitted on : Friday, May 29, 2020 - 11:51:55 AM
Last modification on : Saturday, December 12, 2020 - 8:32:06 AM

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Georges Tawil, Frederic Jamme, Matthieu Refregiers, Anders Vikso-Nielsen, Paul Colonna, et al.. In Situ Tracking of Enzymatic Breakdown of Starch Granules by Synchrotron UV Fluorescence Microscopy. Analytical Chemistry, American Chemical Society, 2011, 83 (3), pp.989 - 993. ⟨10.1021/ac1027512⟩. ⟨hal-02649349⟩

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