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

Authors
  • Leis, Benedikt1
  • Heinze, Simon1
  • Angelov, Angel1
  • Pham, Vu Thuy Trang1
  • Thürmer, Andrea2
  • Jebbar, Mohamed3
  • Golyshin, Peter N4
  • Streit, Wolfgang R5
  • Daniel, Rolf2
  • Liebl, Wolfgang1
  • 1 Department of Microbiology, School of Life Sciences Weihenstephan, Technische Universität München , Freising-Weihenstephan , Germany. , (Germany)
  • 2 Göttingen Genomics Laboratory, Department of Genomic and Applied Microbiology, Georg-August University Göttingen , Göttingen , Germany. , (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. , (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. , (Germany)
Type
Published Article
Journal
Frontiers in Bioengineering and Biotechnology
Publisher
Frontiers Media SA
Publication Date
Jan 01, 2015
Volume
3
Pages
95–95
Identifiers
DOI: 10.3389/fbioe.2015.00095
PMID: 26191525
Source
Medline
Keywords
License
Unknown

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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