Functional metagenomics unveils a multifunctional glycosyl hydrolase from the family 43 catalysing the breakdown of plant polymers in the calf rumen
Vieites, Jose Marıa
Nechitaylo, Taras Y.
Chernikova, Tatyana N.
Golyshina, Olga V.
Golyshin, Peter N.
Contributor(s)Reva, Oleg N.
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AbstractMicrobial communities from cow rumen are known for their ability to degrade diverse plant polymers at high rates. In this
work, we identified 15 hydrolases through an activity-centred metagenome analysis of a fibre-adherent microbial
community from dairy cow rumen. Among them, 7 glycosyl hydrolases (GHs) and 1 feruloyl esterase were successfully
cloned, expressed, purified and characterised. The most striking result was a protein of GH family 43 (GHF43), hereinafter
designated as R_09-02, which had characteristics very distinct from the other proteins in this family with mono-functional bxylosidase,
a-xylanase, a-L-arabinase and a-L-arabinofuranosidase activities. R_09-02 is the first multifunctional enzyme to
exhibit b-1,4 xylosidase, a-1,5 arabinofur(pyr)anosidase, b-1,4 lactase, a-1,6 raffinase, a-1,6 stachyase, b-galactosidase and a-
1,4 glucosidase activities. The R_09-02 protein appears to originate from the chromosome of a member of Clostridia, a class
of phylum Firmicutes, members of which are highly abundant in ruminal environment. The evolution of R_09-02 is
suggested to be driven from the xylose- and arabinose-specific activities, typical for GHF43 members, toward a broader
specificity to the glucose- and galactose-containing components of lignocellulose. The apparent capability of enzymes from
the GHF43 family to utilise xylose-, arabinose-, glucose- and galactose-containing oligosaccharides has thus far been
neglected by, or could not be predicted from, genome and metagenome sequencing data analyses. Taking into account the
abundance of GHF43-encoding gene sequences in the rumen (up to 7% of all GH-genes) and the multifunctional phenotype
herein described, our findings suggest that the ecological role of this GH family in the digestion of ligno-cellulosic matter
should be significantly reconsidered.
The Spanish CDTI (research projects CENIT 2007-1031 and Cenit BioSos, I+DEA), the Ministry of Economy and Competitiveness «Fondo de inversion local para el empleo-Gobierno de Espana». AB thanks the
Spanish MEC for a FPU fellowship. M-EG thanks the CSIC for a JAE-Doc fellowship. TYN, PNG, and OVG acknowledge the support of grant 0313751K from the
Federal Ministry for Science and Education (BMBF) within the GenoMikPlus initiative.
Ferrer M, Ghazi A, Beloqui A, Vieites JM, Lopez-Cortes N, et al. (2012) Functional Metagenomics Unveils a Multifunctional Glycosyl Hydrolase from the Family 43 Catalysing the Breakdown of Plant Polymers in the Calf Rumen. PLoS ONE 7(6): e38134. DOI: 10.1371/journal.pone.0038134.