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Carbohydrate-active enzymes identified by metagenomic analysis of deep-sea sediment bacteria |
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| Authors: | Barbara Klippel Kerstin Sahm Alexander Basner Sigrid Wiebusch Patrick John Ute Lorenz Anke Peters Fumiyoshi Abe Kyoma Takahashi Olaf Kaiser Alexander Goesmann Sebastian Jaenicke Ralf Grote Koki Horikoshi Garabed Antranikian |
| Affiliation: | 1. Institute of Technical Microbiology, Hamburg University of Technology, Kasernenstr. 12, 21073, Hamburg, Germany 2. Extremobiosphere Research Center (XBR), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka, Kanagawa, 237-0061, Japan 3. Department of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin University, 5-10-1 Fuchinobe, Chuo-ku, Sagamihara, 252-5258, Japan 4. Center for Deep Earth Exploration (CDEX), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 3173-25, Showa-machi, Kanazawa-ku, Yokohama, Kanagawa, 236-0001, Japan 5. Roche Diagnostics GmbH, Nonnenwald 2, 82377, Penzberg, Germany 6. Center for Biotechnology, CeBiTec, Universitaetsstrasse 25, 33615, Bielefeld, Germany |
| Abstract: | Subseafloor sediment samples derived from a sediment core of 60 m length were used to enrich psychrophilic aerobic bacteria on cellulose, xylan, chitin, and starch. A variety of species belonging to Alpha- and Gammaproteobacteria and to Flavobacteria were isolated from sediment depths between 12 and 42 mbsf. Metagenomic DNA purified from the pooled enrichments was sequenced and analyzed for phylogenetic composition and presence of genes encoding carbohydrate-active enzymes. More than 200 open reading frames coding for glycoside hydrolases were identified, and more than 60 of them relevant for enzymatic degradation of lignocellulose. Four genes encoding β-glucosidases with less than 52 % identities to characterized enzymes were chosen for recombinant expression in Escherichia coli. In addition one endomannanase, two endoxylanases, and three β-xylosidases were produced recombinantly. All genes could be actively expressed. Functional analysis revealed discrepancies and additional variability for the recombinant enzymes as compared to the sequence-based predictions. |
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