Arabinan saccharification by biogas reactor metagenome-derived arabinosyl hydrolases

Yajing Liu, Angel Angelov, Werner Feiler, Melanie Baudrexl, Vladimir Zverlov, Wolfgang Liebl*, Sonja Vanderhaeghen*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Background: Plant cell walls represent the most plentiful renewable organic resource on earth, but due to their heterogeneity, complex structure and partial recalcitrance, their use as biotechnological feedstock is still limited. Results: In order to identify efficient enzymes for polysaccharide breakdown, we have carried out functional screening of metagenomic fosmid libraries from biogas fermenter microbial communities grown on sugar beet pulp, an arabinan-rich agricultural residue, or other sources containing microbes that efficiently depolymerize polysaccharides, using CPH (chromogenic polysaccharide hydrogel) or ICB (insoluble chromogenic biomass) labeled polysaccharide substrates. Seventy-one depolymerase-encoding genes were identified from 55 active fosmid clones by using Illumina and Sanger sequencing and dbCAN CAZyme (carbohydrate-active enzyme) annotation. An around 56 kb assembled DNA fragment putatively originating from Xylanivirga thermophila strain or a close relative was analyzed in detail. It contained 48 ORFs (open reading frames), of which 31 were assigned to sugar metabolism. Interestingly, a large number of genes for enzymes putatively involved in degradation and utilization of arabinose-containing carbohydrates were found. Seven putative arabinosyl hydrolases from this DNA fragment belonging to glycoside hydrolase (GH) families GH51 and GH43 were biochemically characterized, revealing two with endo-arabinanase activity and four with exo-α-l-arabinofuranosidase activity but with complementary cleavage properties. These enzymes were found to act synergistically and can completely hydrolyze SBA (sugar beet arabinan) and DA (debranched arabinan). Conclusions: We screened 32,776 fosmid clones from several metagenomic libraries with chromogenic lignocellulosic substrates for functional enzymes to advance the understanding about the saccharification of recalcitrant lignocellulose. Seven putative X. thermophila arabinosyl hydrolases were characterized for pectic substrate degradation. The arabinosyl hydrolases displayed maximum activity and significant long-term stability around 50 °C. The enzyme cocktails composed in this study fully degraded the arabinan substrates and thus could serve for arabinose production in food and biofuel industries.

    Original languageEnglish (US)
    Article number121
    JournalBiotechnology for Biofuels and Bioproducts
    Volume15
    Issue number1
    DOIs
    StatePublished - Dec 2022

    Keywords

    • Arabinosyl hydrolases
    • Metagenomics
    • Pentose metabolism
    • Sugar beet pulp degradation
    • Xylanivirga thermophila

    ASJC Scopus subject areas

    • Biotechnology
    • Renewable Energy, Sustainability and the Environment
    • Applied Microbiology and Biotechnology
    • Energy (miscellaneous)
    • Management, Monitoring, Policy and Law

    Fingerprint

    Dive into the research topics of 'Arabinan saccharification by biogas reactor metagenome-derived arabinosyl hydrolases'. Together they form a unique fingerprint.

    Cite this