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Xyloglucan breakdown by endo-xyloglucanase family 74 from Aspergillus fumigatus

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Abstract

Xyloglucan is the most abundant hemicellulose in primary walls of spermatophytes except for grasses. Xyloglucan-degrading enzymes are important in lignocellulosic biomass hydrolysis because they remove xyloglucan, which is abundant in monocot-derived biomass. Fungal genomes encode numerous xyloglucanase genes, belonging to at least six glycoside hydrolase (GH) families. GH74 endo-xyloglucanases cleave xyloglucan backbones with unsubstituted glucose at the −1 subsite or prefer xylosyl-substituted residues in the −1 subsite. In this work, 137 GH74-related genes were detected by examining 293 Eurotiomycete genomes and Ascomycete fungi contained one or no GH74 xyloglucanase gene per genome. Another interesting feature is that the triad of tryptophan residues along the catalytic cleft was found to be widely conserved among Ascomycetes. The GH74 from Aspergillus fumigatus (AfXEG74) was chosen as an example to conduct comprehensive biochemical studies to determine the catalytic mechanism. AfXEG74 has no CBM and cleaves the xyloglucan backbone between the unsubstituted glucose and xylose-substituted glucose at specific positions, along the XX motif when linked to regions deprived of galactosyl branches. It resembles an endo-processive activity, which after initial random hydrolysis releases xyloglucan-oligosaccharides as major reaction products. This work provides insights on phylogenetic diversity and catalytic mechanism of GH74 xyloglucanases from Ascomycete fungi.

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Acknowledgements

We are grateful to the State of São Paulo Research Foundation (FAPESP) for its financial support (2012/20549-4 to ARLD; 2008/58037-9 to FMS; 2014/18714-2 to FS). ARLD also received a FAPESP fellowship (2011/02169-7). This work was also financially supported by CNPq: 442333/2014-5, 310186/2014-5, 304445/2014-2, and 441912/2014-1. APS and MSB thank the Instituto Nacional de Ciência e Tecnologia do Bioetanol – INCT do Bioetanol (FAPESP 2008/57908-6 and CNPq 574002/2008-1). We would like to thank the entire team of the molecular biology laboratory from CTBE/CNPEM.

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Authors and Affiliations

  1. Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil

    André Ricardo de Lima Damasio, Marcelo Ventura Rubio, Thiago Augusto Gonçalves, Gabriela Felix Persinoti, Fernando Segato, Fabiano Jares Contesini & Fabio Marcio Squina

  2. Department of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas, Campinas, SP, Brazil

    André Ricardo de Lima Damasio, Marcelo Ventura Rubio & Thiago Augusto Gonçalves

  3. Departamento de Biotecnologia, Escola de Engenharia de Lorena (EEL), Universidade de São Paulo (USP), Lorena, SP, Brazil

    Fernando Segato

  4. Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA

    Rolf Alexander Prade

  5. Laboratório de Fisiologia e Ecologia de Plantas (LAFIECO), Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo (USP), São Paulo, SP, Brazil

    Amanda Pereira de Souza & Marcos Silveira Buckeridge

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  1. André Ricardo de Lima Damasio
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  2. Marcelo Ventura Rubio
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  3. Thiago Augusto Gonçalves
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Correspondence to Fabio Marcio Squina.

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Damasio, A.R., Rubio, M.V., Gonçalves, T.A. et al. Xyloglucan breakdown by endo-xyloglucanase family 74 from Aspergillus fumigatus . Appl Microbiol Biotechnol 101, 2893–2903 (2017). https://doi.org/10.1007/s00253-016-8014-6

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