Additionally, selleck chem several GT, PL and CE domains were identi fied as relevant. These CAZy families, as well as GH115 and CBM56, are not included in Figure 2, as they are not annotated for all sequences. Identification of plant biomass degraders from a cow rumen metagenome We used our method to predict the plant biomass degrading capabilities for 15 draft genomes of uncul tured microbes reconstructed from the metagenome of a microbial community adherent to switchgrass in cow rumen. The draft genomes repre sent genomes with more than 50% of the sequence reconstructed by taxonomic binning of the metagenome sample. The microbial community adherent to switch grass is likely to be enriched with plant biomass degraders, as it was found to differ from the rumen fluid community in its taxonomic composition and degrad ation of switch grass after incubation in cow rumen had occurred.

For identification of plant biomass degrading microbes, we classified each draft genome individually with the eSVMbPFAM and eSVMCAZY B models, which had the highest macro accuracy based on Pfam domain or CAZy family annotations, respectively. The eSVMbPFAM classifier assigned seven of the draft genomes to plant bio mass degraders. One of these, genome APb, was found by 16S rRNA analysis to be related to the fibrolytic species Butyrivibrio fibrisolvens. Four others are of the order of Bacteroidales, and include all but one draft genomes affiliated to the Bacteroidales. The 6th and 7th predicted degrader, represented by gen ome AIa and AWa, belong to the Clostridiales, like genome APb.

The eSVMCAZY B classifier also assigned five of these genomes to the plant biomass degraders. Add itionally it classified genome AH as plant biomass degrading, while being ambiguous in the assignment of AFa. To validate these predictions, we searched the draft genomes for genes encoding 51 enzymatically active glycoside hydrolases AV-951 characterized from the same rumen dataset. Genomes AGa, AC2a, AJ and AIa were all linked to different enzymes of varying specificities. AC2a was linked to cellulose deg radation, specifically to a carboxymethyl cellulose degrading GH5 endoglucanase as well as GH9 enzyme capable of degrading insoluble cellulosic substrates such as AvicelW. AIa demonstrated capabilities towards xylan and soluble cellulosic substrates with affiliations to four GH10 xylanases. Both AGa and AJ demonstrated broader substrate versatility and were linked to enzymes Dorsomorphin ALK with capabilities towards cellulosic substrates CMC and AvicelW, hemicellulosic substrates lichenan and xylan, as well as the natural feedstocks miscanthus and switchgrass. Import antly, no carbohydrate active enzymes were affiliated to draft genomes that were predicted to not possess plant biomass degrading capabilities.