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.
Journal
Microbial biotechnology
ISSN: 1751-7915
Titre abrégé: Microb Biotechnol
Pays: United States
ID NLM: 101316335
Informations de publication
Date de publication:
Jun 2024
Jun 2024
Historique:
revised:
12
03
2024
received:
26
01
2024
accepted:
20
03
2024
medline:
3
6
2024
pubmed:
3
6
2024
entrez:
3
6
2024
Statut:
ppublish
Résumé
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
Identifiants
pubmed: 38829370
doi: 10.1111/1751-7915.14466
doi:
Substances chimiques
Detergents
0
Cellulase
EC 3.2.1.4
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e14466Subventions
Organisme : Danmarks Frie Forskningsfond
ID : 0217-00052B
Organisme : Fundação de Amparo à Pesquisa do Estado de São Paulo
ID : 2019/06663-8
Organisme : Fundação de Amparo à Pesquisa do Estado de São Paulo
ID : 2022/01756-0
Organisme : Novo Nordisk Fonden
ID : NNFSA170028392
Informations de copyright
© 2024 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd.
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