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Functional Annotation of Fibrobacter succinogenes S85 Carbohydrate Active Enzymes

Permanent URL:
http://handle.nal.usda.gov/10113/53697
Abstract:
Fibrobacter succinogenes is a cellulolytic bacterium that degrades plant cell wall biomass in ruminant animals and is among the most rapidly fibrolytic of all mesophilic bacteria. The complete genome sequence of Fisuc was completed by the DOE Joint Genome Institute in late 2009. Using new expression tools developed at Lucigen and C5-6 Technologies and a multi-substrate screen, 5,760 random shotgun expression clones were screened for biomass-degrading enzymes, representing 2× genome expression coverage. From the screen, 169 positive hits were recorded and 33 were unambiguously identified by sequence analysis of the inserts as belonging to CAZy family genes. Eliminating duplicates, 24 unique CAZy genes were found by functional screening. Several previously uncharacterized enzymes were discovered using this approach and a number of potentially mis-annotated enzymes were functionally characterized. To complement this approach, a high-throughput system was developed to clone and express all the annotated glycosyl hydrolases and carbohydrate esterases in the genome. Using this method, six previously described and five novel CAZy enzymes were cloned, expressed, and purified in milligram quantities.
Author(s):
Brumm, Phillip , Mead, David , Boyum, Julie , Drinkwater, Colleen , Gowda, Krishne , Stevenson, David , Weimer, Paul
Subject(s):
Fibrobacter succinogenes , animals , bacteria , biodegradation , biomass , carbohydrates , cell walls , clones , esterases , genes , plants , rumen microorganisms , screening , sequence analysis
Note:
Includes references
Source:
Applied biochemistry and biotechnology 2011 Mar., v. 163, no. 5
Language:
English
Publisher:
Humana Press Inc
Year:
2011
Collection:
Journal Articles, USDA Authors, Peer-Reviewed
File:
Download [PDF File]
Rights:
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.