High-throughput generation of product profiles for arabinoxylan-active enzymes from metagenomes.

High-throughput generation of product profiles for arabinoxylan-active enzymes from metagenomes. Appl Environ Microbiol. 2020 Sep 18;: Authors: Fonseca MJMD, Armstrong Z, Withers SG, Briers Y Abstract Metagenomics is an exciting alternative to seek for carbohydrate-active enzymes from a range of sources. Typically, metagenomics reveals dozens of putative catalysts that require functional characterization for further application in industrial processes. High-throughput screening methods compatible with adequate natural substrates are crucial for an accurate functional elucidation of substrate preferences. Based on DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) analysis of enzymatic reaction products, we generated product profiles to consequently infer substrate cleavage positions, resulting in the generation of enzymatic degradation maps. Product profiles were produced in high-throughput for arabinoxylan (AX)-active enzymes belonging to the glycoside hydrolase families GH43 (subfamilies 2 (MG432), 7 (MG437) and 28 (MG4328)) and GH8 (MG8) starting from twelve (arabino)xylo-oligosaccharides. These enzymes were discovered through functional metagenomic studies of faeces from the North American beaver (Castor canadensis). This work shows how enzyme loading alters the product profiles produced by all enzymes studied and gives insight into AX degradation patterns revealing sequential substrate preferences o...
Source: Applied and Environmental Microbiology - Category: Microbiology Authors: Tags: Appl Environ Microbiol Source Type: research