title: Carbohydrate-active enzymes identified by metagenomic analysis of deep-sea sediment bacteria.
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.
authors.affiliation: Institute of Technical Microbiology, Hamburg University of Technology, Kasernenstr. 12, 21073, Hamburg, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Klippel
authors.forename: Kerstin
authors.initials: K
authors.lastname: Sahm
authors.forename: Alexander
authors.initials: A
authors.lastname: Basner
authors.forename: Sigrid
authors.initials: S
authors.lastname: Wiebusch
authors.forename: Patrick
authors.initials: P
authors.lastname: John
authors.forename: Ute
authors.initials: U
authors.lastname: Lorenz
authors.forename: Anke
authors.initials: A
authors.lastname: Peters
authors.forename: Fumiyoshi
authors.initials: F
authors.lastname: Abe
authors.forename: Kyoma
authors.initials: K
authors.lastname: Takahashi
authors.forename: Olaf
authors.initials: O
authors.lastname: Kaiser
authors.forename: Alexander
authors.initials: A
authors.lastname: Goesmann
authors.forename: Sebastian
authors.initials: S
authors.lastname: Jaenicke
authors.forename: Ralf
authors.initials: R
authors.lastname: Grote
authors.forename: Koki
authors.initials: K
authors.lastname: Horikoshi
authors.forename: Garabed
authors.initials: G
authors.lastname: Antranikian
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D044602:Cellulases / Q000235:genetics* / Q000378:metabolism; D041965:Flavobacteriaceae / Q000201:enzymology / Q000235:genetics*; D020563:Gammaproteobacteria / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D019015:Geologic Sediments / Q000382:microbiology*; D054892:Metagenome; D012623:Seawater / Q000382:microbiology; D014995:Xylosidases / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D044602:Cellulases; D014995:Xylosidases
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