Biochemical characterization of a novel monospecific endo-β-1,4-glucanase belonging to GH Family 5 from a rhizosphere metagenomic library

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dc.contributor.author Wierzbicka-Woś, Anna
dc.contributor.author Henneberger, Ruth
dc.contributor.author Batista-García, Ramón Alberto
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 2019-11-20T05:46:24Z
dc.date.available 2019-11-20T05:46:24Z
dc.date.issued 2019-06-14
dc.identifier.citation Wierzbicka-Wos, A., Henneberger, R., Batista-García, R.A., Martinez-Avila, L., Jackson, S.A., Kennedy, J. and Dobson, A.D. (2019) 'Biochemical characterization of a novel monospecific endo-Beta-1, 4-glucanase belonging to GH family 5 from a rhizosphere metagenomic library.' Frontiers in Microbiology, 10, 1342. (19pp.) doi:10.3389/fmicb.2019.01342 en
dc.identifier.volume 10 en
dc.identifier.startpage 1 en
dc.identifier.endpage 19 en
dc.identifier.uri http://hdl.handle.net/10468/9133
dc.identifier.doi 10.3389/fmicb.2019.01342 en
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 modelling 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 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 in presence of 10 mM ions Zn2+, Ni2+, Co2+, Fe3+ and reagents such as 1 M guanidine HCl, 0.1% sodium dodecyl sulphate (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
dc.description.sponsorship Federation of European Microbiological Societies (FEMSRG-2014-0007.R1); CONACyT-SEP (285816); Science Foundation Ireland (SSPC-2, 12/RC/2275) en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Frontiers Media en
dc.relation.uri https://www.frontiersin.org/articles/10.3389/fmicb.2019.01342
dc.rights © 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. © 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. 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) and the copyright owner(s) 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. en
dc.rights.uri https://creativecommons.org/licenses/by/4.0/ en
dc.subject Cellulase en
dc.subject Endo-glucanase en
dc.subject Glycosyl hydrolase (GH) en
dc.subject Functional metagenomics en
dc.subject Biochemical characterisation en
dc.title Biochemical characterization of a novel monospecific endo-β-1,4-glucanase belonging to GH Family 5 from a rhizosphere metagenomic library en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Stephen Jackson, Environmental Research Institute, University College Cork, Cork, Ireland. +353-21-490-3000 Email: sjackson@ucc.ie en
dc.internal.availability Full text available en
dc.description.version Published Version en
dc.contributor.funder Federation of European Microbiological Societies en
dc.contributor.funder Science Foundation Ireland en
dc.contributor.funder Consejo Nacional de Ciencia y Tecnología en
dc.description.status Peer reviewed en
dc.identifier.journaltitle Frontiers in Microbiology en
dc.internal.IRISemailaddress sjackson@ucc.ie en
dc.identifier.articleid 1342 en
dc.identifier.eissn 1664-302X


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© 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. © 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. 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) and the copyright owner(s) 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. Except where otherwise noted, this item's license is described as © 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. © 2019 Wierzbicka-Woś, Henneberger, Batista-García, Martínez-Ávila, Jackson, Kennedy and Dobson. 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) and the copyright owner(s) 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.
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