Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
Type | Article | ||||||||||||||||
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Date | 2015-07 | ||||||||||||||||
Language | English | ||||||||||||||||
Author(s) | Leis Benedikt1, Heinze Simon1, Angelov Angel1, Pham Vu Thuy Trang1, Thürmer Andrea2, Jebbar Mohamed3, Golyshin Peter N.4, Streit Wolfgang R.5, Daniel Rolf2, Liebl Wolfgang1 | ||||||||||||||||
Affiliation(s) | 1 : Department of Microbiology, School of Life Sciences Weihenstephan, Technische Universität München, Freising- Weihenstephan, German 2 : Göttingen Genomics Laboratory, Department of Genomic and Applied Microbiology, Georg- August University Göttingen, Göttingen, Germany 3 : Laboratoire de Microbiologie des Environnements Extrêmes-UMR 6197 (CNRS-Ifremer-UBO), Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, Plouzané, France 4 : School of Biological Sciences, Bangor University, Bangor, UK 5 : Fakultät für Mathematik, Informatik und Naturwissenschaften Biologie, Biozentrum Klein Flottbek, Universität Hamburg, Hamburg, Germany |
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Source | Frontiers in Bioengineering and Biotechnology (2296-4185) (Frontiers Media SA), 2015-07 , Vol. 3 , N. 95 , P. 10p. | ||||||||||||||||
DOI | 10.3389/fbioe.2015.00095 | ||||||||||||||||
WOS© Times Cited | 23 | ||||||||||||||||
Keyword(s) | functional screenings, extreme thermostable protein, archaeal endoglucanase, enzymatic characterization | ||||||||||||||||
Abstract | Extreme habitats serve as a source of enzymes that are active under extreme conditions and are candidates for industrial applications. In this work, six large-insert mixed genomic libraries were screened for hydrolase activities in a broad temperature range (8–70°C). Among a variety of hydrolytic activities, one fosmid clone, derived from a library of pooled isolates of hyperthermophilic archaea from deep sea vents, displayed hydrolytic activity on carboxymethyl cellulose substrate plates at 70°C but not at lower temperatures. Sequence analysis of the fosmid insert revealed a gene encoding a novel glycoside hydrolase family 12 (GHF12) endo-1,4-β-glucanase, termed Cel12E. The enzyme shares 45% sequence identity with a protein from the archaeon Thermococcus sp. AM4 and displays a unique multidomain architecture. Biochemical characterization of Cel12E revealed a remarkably thermostable protein, which appears to be of archaeal origin. The enzyme displayed maximum activity at 92°C and was active on a variety of linear 1,4-β-glucans like carboxymethyl cellulose, β-glucan, lichenan, and phosphoric acid swollen cellulose. The protein is able to bind to various insoluble β-glucans. Product pattern analysis indicated that Cel12E is an endo-cleaving β-glucanase. Cel12E expands the toolbox of hyperthermostable archaeal cellulases with biotechnological potential. |
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