Open AccessActivity-Based Metagenomic Screening and Biochemical Characterization of Bovine Ruminal Protozoan Glycoside Hydrolases
Author(s) -
Seth D. Findley,
Melanie R. Mormile,
Andrea Sommer-Hurley,
Xuecheng Zhang,
Peter A. Tipton,
Krista L. Arnett,
Julie Porter,
M. S. Kerley,
Gary Stacey
Publication year - 2011
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.05925-11
Subject(s) - rumen , biology , metagenomics , axenic , ruminant , complementary dna , gene , cdna library , glycoside hydrolase , biochemistry , fermentation , microbiology and biotechnology , bacteria , genetics , pasture , ecology
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.