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Issue 8, 2017
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High-throughput analysis of endogenous fruit glycosyl hydrolases using a novel chromogenic hydrogel substrate assay

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Abstract

A broad range of enzyme activities can be found in a wide range of different fruits and fruiting bodies but there is a lack of methods where many samples can be handled in a high-throughput and efficient manner. In particular, plant polysaccharide degrading enzymes – glycosyl hydrolases (GHs) play an important role in fruit development and ripening processes by modulating the plant cell wall. Knowledge about these enzymes is important for research in fruit development and also important for industry regarding postharvest properties. Although advances in genetic control and cell wall biochemistry have led to a more profound understanding of the importance of GH activity and regulation, current methods for determining glycosyl hydrolase activity are lacking in throughput and fail to keep up with data output from transcriptome research. Here we present the use of a versatile, easy-to-handle, multiplexed and highly reproducible method using CPH assays where different fruits have been screened for enzyme activity. Additionally, the importance and impact of the extraction method and buffer conditions on the assay are investigated. We will show that one experimental setup can be used for testing all enzymes.

Graphical abstract: High-throughput analysis of endogenous fruit glycosyl hydrolases using a novel chromogenic hydrogel substrate assay

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Supplementary files

Article information


Submitted
23 Dec 2016
Accepted
05 Feb 2017
First published
06 Feb 2017

Anal. Methods, 2017,9, 1242-1247
Article type
Communication

High-throughput analysis of endogenous fruit glycosyl hydrolases using a novel chromogenic hydrogel substrate assay

J. Schückel, S. K. Kračun, T. F. Lausen, W. G. Tycho Willats and B. Jørgensen, Anal. Methods, 2017, 9, 1242
DOI: 10.1039/C6AY03431D

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