title: Activity-based metagenomic screening and biochemical characterization of bovine ruminal protozoan glycoside hydrolases.
abstract: The rumen, the foregut of herbivorous ruminant animals such as cattle, functions as a bioreactor to process complex plant material. Among the numerous and diverse microbes involved in ruminal digestion are the ruminal protozoans, which are single-celled, ciliated eukaryotic organisms. An activity-based screen was executed to identify genes encoding fibrolytic enzymes present in the metatranscriptome of a bovine ruminal protozoan-enriched cDNA expression library. Of the four novel genes identified, two were characterized in biochemical assays. Our results provide evidence for the effective use of functional metagenomics to retrieve novel enzymes from microbial populations that cannot be maintained in axenic cultures.
authors.affiliation: Center for Sustainable Energy, National Center for Soybean Biotechnology, Division of Plant Sciences, University of Missouri, Columbia, Missouri, USA.
authors.forename: Seth D
authors.initials: SD
authors.lastname: Findley
authors.forename: Melanie R
authors.initials: MR
authors.lastname: Mormile
authors.forename: Andrea
authors.initials: A
authors.lastname: Sommer-Hurley
authors.forename: Xue-Cheng
authors.initials: XC
authors.lastname: Zhang
authors.forename: Peter
authors.initials: P
authors.lastname: Tipton
authors.forename: Krista
authors.initials: K
authors.lastname: Arnett
authors.forename: James H
authors.initials: JH
authors.lastname: Porter
authors.forename: Monty
authors.initials: M
authors.lastname: Kerley
authors.forename: Gary
authors.initials: G
authors.lastname: Stacey
mesh_terms: D000818:Animals; D002417:Cattle; D016000:Cluster Analysis; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism*; D054892:Metagenome; D008969:Molecular Sequence Data; D010802:Phylogeny; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism*; D012417:Rumen / Q000469:parasitology*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
chemical_list: D016054:DNA, Protozoan; D015800:Protozoan Proteins; D006026:Glycoside Hydrolases