Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland
Abstract Microbial communities from extreme environments are largely understudied, but are essential as producers of metabolites, including enzymes, for industrial processes. As cultivation of most microorganisms remains a challenge, culture‐independent approaches for enzyme discovery in the form of...
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Online Access: | http://dx.doi.org/10.1111/1751-7915.14466 |
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crwiley:10.1111/1751-7915.14466 2024-09-15T18:09:33+00:00 Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland Oliva, Bianca Zervas, Athanasios Stougaard, Peter Westh, Peter Thøgersen, Mariane Schmidt Danmarks Frie Forskningsfond Fundação de Amparo à Pesquisa do Estado de São Paulo Novo Nordisk Fonden 2024 http://dx.doi.org/10.1111/1751-7915.14466 en eng Wiley http://creativecommons.org/licenses/by-nc-nd/4.0/ Microbial Biotechnology volume 17, issue 6 ISSN 1751-7915 1751-7915 journal-article 2024 crwiley https://doi.org/10.1111/1751-7915.14466 2024-07-02T04:14:04Z Abstract Microbial communities from extreme environments are largely understudied, but are essential as producers of metabolites, including enzymes, for industrial processes. As cultivation of most microorganisms remains a challenge, culture‐independent approaches for enzyme discovery in the form of metagenomics to analyse the genetic potential of a community are rapidly becoming the way forward. This study focused on analysing a metagenome from the cold and alkaline ikaite columns in Greenland, identifying 282 open reading frames (ORFs) that encoded putative carbohydrate‐modifying enzymes with potential applications in, for example detergents and other processes where activity at low temperature and high pH is desired. Seventeen selected ORFs, representing eight enzyme families were synthesized and expressed in two host organisms, Escherichia coli and Aliivibrio wodanis . Aliivibrio wodanis demonstrated expression of a more diverse range of enzyme classes compared to E. coli , emphasizing the importance of alternative expression systems for enzymes from extremophilic microorganisms. To demonstrate the validity of the screening strategy, we chose a recombinantly expressed cellulolytic enzyme from the metagenome for further characterization. The enzyme, Cel240, exhibited close to 40% of its relative activity at low temperatures (4°C) and demonstrated endoglucanase characteristics, with a preference for cellulose substrates. Despite low sequence similarity with known enzymes, computational analysis and structural modelling confirmed its cellulase‐family affiliation. Cel240 displayed activity at low temperatures and good stability at 25°C, activity at alkaline pH and increased activity in the presence of CaCl 2 , making it a promising candidate for detergent and washing industry applications. Article in Journal/Newspaper Greenland Wiley Online Library Microbial Biotechnology 17 6 |
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Wiley Online Library |
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English |
description |
Abstract Microbial communities from extreme environments are largely understudied, but are essential as producers of metabolites, including enzymes, for industrial processes. As cultivation of most microorganisms remains a challenge, culture‐independent approaches for enzyme discovery in the form of metagenomics to analyse the genetic potential of a community are rapidly becoming the way forward. This study focused on analysing a metagenome from the cold and alkaline ikaite columns in Greenland, identifying 282 open reading frames (ORFs) that encoded putative carbohydrate‐modifying enzymes with potential applications in, for example detergents and other processes where activity at low temperature and high pH is desired. Seventeen selected ORFs, representing eight enzyme families were synthesized and expressed in two host organisms, Escherichia coli and Aliivibrio wodanis . Aliivibrio wodanis demonstrated expression of a more diverse range of enzyme classes compared to E. coli , emphasizing the importance of alternative expression systems for enzymes from extremophilic microorganisms. To demonstrate the validity of the screening strategy, we chose a recombinantly expressed cellulolytic enzyme from the metagenome for further characterization. The enzyme, Cel240, exhibited close to 40% of its relative activity at low temperatures (4°C) and demonstrated endoglucanase characteristics, with a preference for cellulose substrates. Despite low sequence similarity with known enzymes, computational analysis and structural modelling confirmed its cellulase‐family affiliation. Cel240 displayed activity at low temperatures and good stability at 25°C, activity at alkaline pH and increased activity in the presence of CaCl 2 , making it a promising candidate for detergent and washing industry applications. |
author2 |
Danmarks Frie Forskningsfond Fundação de Amparo à Pesquisa do Estado de São Paulo Novo Nordisk Fonden |
format |
Article in Journal/Newspaper |
author |
Oliva, Bianca Zervas, Athanasios Stougaard, Peter Westh, Peter Thøgersen, Mariane Schmidt |
spellingShingle |
Oliva, Bianca Zervas, Athanasios Stougaard, Peter Westh, Peter Thøgersen, Mariane Schmidt Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
author_facet |
Oliva, Bianca Zervas, Athanasios Stougaard, Peter Westh, Peter Thøgersen, Mariane Schmidt |
author_sort |
Oliva, Bianca |
title |
Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
title_short |
Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
title_full |
Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
title_fullStr |
Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
title_full_unstemmed |
Metagenomic exploration of cold‐active enzymes for detergent applications: Characterization of a novel, cold‐active and alkali‐stable GH8 endoglucanase from ikaite columns in SW Greenland |
title_sort |
metagenomic exploration of cold‐active enzymes for detergent applications: characterization of a novel, cold‐active and alkali‐stable gh8 endoglucanase from ikaite columns in sw greenland |
publisher |
Wiley |
publishDate |
2024 |
url |
http://dx.doi.org/10.1111/1751-7915.14466 |
genre |
Greenland |
genre_facet |
Greenland |
op_source |
Microbial Biotechnology volume 17, issue 6 ISSN 1751-7915 1751-7915 |
op_rights |
http://creativecommons.org/licenses/by-nc-nd/4.0/ |
op_doi |
https://doi.org/10.1111/1751-7915.14466 |
container_title |
Microbial Biotechnology |
container_volume |
17 |
container_issue |
6 |
_version_ |
1810447119791685632 |