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dc.contributor.author
Wierzbicka-Wos, Anna
dc.contributor.author
Henneberger, Ruth
dc.contributor.author
Alberto Batista-García, Ramón
dc.contributor.author
Martínez-Ávila, Liliana
dc.contributor.author
Jackson, Stephen A.
dc.contributor.author
Kennedy, Jonathan
dc.contributor.author
Dobson, Alan D.W.
dc.date.accessioned
2019-06-28T09:52:35Z
dc.date.available
2019-06-28T07:16:42Z
dc.date.available
2019-06-28T09:52:35Z
dc.date.issued
2019-06-14
dc.identifier.issn
1664-302X
dc.identifier.other
10.3389/fmicb.2019.01342
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/350289
dc.identifier.doi
10.3929/ethz-b-000350289
dc.description.abstract
Cellulases have a broad range of different industrial applications, ranging from food and beverages to pulp and paper and the biofuels area. Here a metagenomics based strategy was used to identify the cellulolytic enzyme CelRH5 from the rhizosphere. CelRH5 is a novel monospecific endo-β-1,4-glucanase belonging to the glycosyl hydrolase family 5 (GH5). Structural based modeling analysis indicated that CelRH5 is related to endo-β-1,4-glucanases derived from thermophilic microorganisms such as Thermotoga maritima, Fervidobacterium nodosum, and Ruminiclostridium thermocellum sharing 30-40% amino acid sequence identity. The molecular weight of the enzyme was determined as 40.5 kDa. Biochemical analyses revealed that the enzyme displayed good activity with soluble forms of cellulose as a substrate such as ostazin brilliant red hydroxyethyl cellulose (OBR-HEC), carboxymethylcellulose (CMC), hydroxyethyl cellulose (HEC), and insoluble azurine cross-linked hydroxyethylcellulose (AZCL-HEC). The enzyme shows highest enzymatic activity at pH 6.5 with high pH tolerance, remaining stable in the pH range 4.5–8.5. Highest activity was observed at 40°C, but CelRH5 is psychrotolerant being active and stable at temperatures below 30°C. The presence of the final products of cellulose hydrolysis (glucose and cellobiose) or metal ions such as Na+, K+, Li+, and Mg2+, as well as ethylenediaminetetraacetic acid (EDTA), urea, dithiothreitol (DTT), dimethyl sulfoxide (DMSO), 2-mercaptoethanol (2-ME) or glycerol, did not have a marked effect on CelRH5 activity. However, the enzyme is quite sensitive to the presence of 10 mM ions Zn2+, Ni2+, Co2+, Fe3+ and reagents such as 1 M guanidine HCl, 0.1% sodium dodecyl sulfate (SDS) and 20% ethanol. Given that it is psychrotolerant and retains activity in the presence of final cellulose degradation products, metal ions and various reagents, which are common in many technological processes; CelRH5 may be potential suitability for a variety of different biotechnological applications.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Frontiers Media
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject
Functional metagenomics
en_US
dc.subject
Cellulases
en_US
dc.subject
Glycosyl hydrolase family 5
en_US
dc.subject
Endoglucanase
en_US
dc.subject
Psychrotolerant cellulolytic enzyme
en_US
dc.title
Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
ethz.journal.title
Frontiers in Microbiology
ethz.journal.volume
10
en_US
ethz.journal.abbreviated
Front Microbiol
ethz.pages.start
1342
en_US
ethz.size
19 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.publication.place
Lausanne
ethz.publication.status
published
en_US
ethz.date.deposited
2019-06-28T07:16:48Z
ethz.source
WOS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2019-06-28T09:52:47Z
ethz.rosetta.lastUpdated
2024-02-02T08:24:15Z
ethz.rosetta.versionExported
true
ethz.COinS
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