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

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 ac...

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Autores principales: Leis, Benedikt, Heinze, Simon, Angelov, Angel, Pham, Vu Thuy Trang, Thürmer, Andrea, Jebbar, Mohamed, Golyshin, Peter N., Streit, Wolfgang R., Daniel, Rolf, Liebl, Wolfgang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4486847/
https://www.ncbi.nlm.nih.gov/pubmed/26191525
http://dx.doi.org/10.3389/fbioe.2015.00095
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author Leis, Benedikt
Heinze, Simon
Angelov, Angel
Pham, Vu Thuy Trang
Thürmer, Andrea
Jebbar, Mohamed
Golyshin, Peter N.
Streit, Wolfgang R.
Daniel, Rolf
Liebl, Wolfgang
author_facet Leis, Benedikt
Heinze, Simon
Angelov, Angel
Pham, Vu Thuy Trang
Thürmer, Andrea
Jebbar, Mohamed
Golyshin, Peter N.
Streit, Wolfgang R.
Daniel, Rolf
Liebl, Wolfgang
author_sort Leis, Benedikt
collection PubMed
description 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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spelling pubmed-44868472015-07-17 Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon Leis, Benedikt Heinze, Simon Angelov, Angel Pham, Vu Thuy Trang Thürmer, Andrea Jebbar, Mohamed Golyshin, Peter N. Streit, Wolfgang R. Daniel, Rolf Liebl, Wolfgang Front Bioeng Biotechnol Bioengineering and Biotechnology 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. Frontiers Media S.A. 2015-07-01 /pmc/articles/PMC4486847/ /pubmed/26191525 http://dx.doi.org/10.3389/fbioe.2015.00095 Text en Copyright © 2015 Leis, Heinze, Angelov, Pham, Thürmer, Jebbar, Golyshin, Streit, Daniel and Liebl. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Leis, Benedikt
Heinze, Simon
Angelov, Angel
Pham, Vu Thuy Trang
Thürmer, Andrea
Jebbar, Mohamed
Golyshin, Peter N.
Streit, Wolfgang R.
Daniel, Rolf
Liebl, Wolfgang
Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title_full Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title_fullStr Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title_full_unstemmed Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title_short Functional Screening of Hydrolytic Activities Reveals an Extremely Thermostable Cellulase from a Deep-Sea Archaeon
title_sort functional screening of hydrolytic activities reveals an extremely thermostable cellulase from a deep-sea archaeon
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4486847/
https://www.ncbi.nlm.nih.gov/pubmed/26191525
http://dx.doi.org/10.3389/fbioe.2015.00095
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