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Leis B, Heinze S, Angelov A, et al. Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon. Front Bioeng Biotechnol. 2015;3:95doi: 10.3389/fbioe.2015.00095.
Leis, B., Heinze, S., Angelov, A., Pham, V. T., Thürmer, A., Jebbar, M., Golyshin, P. N., Streit, W. R., Daniel, R., & Liebl, W. (2015). Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon. Frontiers in bioengineering and biotechnology, 395. https://doi.org/10.3389/fbioe.2015.00095
Leis, Benedikt, et al. "Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon." Frontiers in bioengineering and biotechnology vol. 3 (2015): 95. doi: https://doi.org/10.3389/fbioe.2015.00095
Leis B, Heinze S, Angelov A, Pham VT, Thürmer A, Jebbar M, Golyshin PN, Streit WR, Daniel R, Liebl W. Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon. Front Bioeng Biotechnol. 2015 Jul 01;3:95. doi: 10.3389/fbioe.2015.00095. eCollection 2015. PMID: 26191525; PMCID: PMC4486847.
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Front Bioeng Biotechnol. 2015 Jul 01;3:95. doi: 10.3389/fbioe.2015.00095. eCollection 2015.

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

Frontiers in bioengineering and biotechnology

Benedikt Leis, Simon Heinze, Angel Angelov, Vu Thuy Trang Pham, Andrea Thürmer, Mohamed Jebbar, Peter N Golyshin, Wolfgang R Streit, Rolf Daniel, Wolfgang Liebl

Affiliations

  1. Department of Microbiology, School of Life Sciences Weihenstephan, Technische Universität München , Freising-Weihenstephan , Germany.
  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.

PMID: 26191525 PMCID: PMC4486847 DOI: 10.3389/fbioe.2015.00095

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.

Keywords: archaeal endoglucanase; enzymatic characterization; extreme thermostable protein; functional screenings

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