Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon

Type Article
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
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 21
Keyword(s) functional screenings, extreme thermostable protein, archaeal endoglucanase, enzymatic characterization

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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Leis Benedikt, Heinze Simon, Angelov Angel, Pham Vu Thuy Trang, Thürmer Andrea, Jebbar Mohamed, Golyshin Peter N., Streit Wolfgang R., Daniel Rolf, Liebl Wolfgang (2015). Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon. Frontiers in Bioengineering and Biotechnology, 3(95), 10p. Publisher's official version : , Open Access version :