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α-Fucosidases with different substrate specificities from two species of Fusarium

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Abstract

Two fungal-secreted α-fucosidases and their genes were characterized. FoFCO1 was purified from culture filtrates of Fusarium oxysporum strain 0685 grown on l-fucose and its encoding gene identified in the sequenced genome of strain 4287. FoFCO1 was active on p-nitrophenyl-α-fucoside (pNP-Fuc), but did not defucosylate a nonasaccharide (XXFG) fragment of pea xyloglucan. A putative α-fucosidase gene (FgFCO1) from Fusarium graminearum was expressed in Pichia pastoris. FgFCO1 was ∼1,800 times less active on pNP-Fuc than FoFCO1, but was able to defucosylate the XXFG nonasaccharide. Although FgFCO1 and FoFCO1 both belong to Glycosyl Hydrolase family 29, they share <25 % overall amino acid identity. Alignment of all available fungal orthologs of FoFCO1 and FgFCO1 indicated that these two proteins belong to two subfamilies of fungal GH29 α-fucosidases. Fungal orthologs of subfamily 1 (to which FoFCO1 belongs) are taxonomically more widely distributed than subfamily 2 (FgFCO1), but neither was universally present in the sequenced fungal genomes. Trichoderma reesei and most species of Aspergillus lack genes for either GH29 subfamily.

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Acknowledgments

This work was funded in part by the U.S. Department of Energy Great Lakes Bioenergy Research Center (DOE Office of Science BER DE-FC02-07ER64494) and in part by grant DE-FG02-91ER200021 to the MSU-Plant Research Laboratory from the U.S. Department of Energy, Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences and Biosciences. Research in A.F.’s laboratory was supported in part by National Research Initiative Competitive Grants 2008-35318-04563 and 2008-35318-04572 from the United States Department of Agriculture National Institute of Food and Agriculture. We thank Ken Keegstra (Michigan State University and DOE-Great Lakes Bioenergy Research Center) for the gift of Arabidopsis FUT1, Cliff Foster (Great Lakes Bioenergy Research Center) for cell wall analysis, and Doug Whitten (Michigan State University Research Technology Support Facility) for proteomics.

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Correspondence to Jonathan D. Walton.

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Paper, J.M., Scott-Craig, J.S., Cavalier, D. et al. α-Fucosidases with different substrate specificities from two species of Fusarium . Appl Microbiol Biotechnol 97, 5371–5380 (2013). https://doi.org/10.1007/s00253-012-4423-3

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