Activity-based metagenomic screening and biochemical characterization of bovine ruminal protozoan glycoside hydrolases

基于活性的牛瘤胃原生动物糖苷水解酶的宏基因组筛选和生化表征

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作者:Seth D Findley, Melanie R Mormile, Andrea Sommer-Hurley, Xue-Cheng Zhang, Peter Tipton, Krista Arnett, James H Porter, Monty Kerley, Gary Stacey

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.

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