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.authorWierzbicka-Woś, Anna
dc.contributor.authorHenneberger, Ruth
dc.contributor.authorBatista-García, Ramón Alberto
dc.contributor.authorMartínez-Ávila, Liliana
dc.contributor.authorJackson, Stephen A.
dc.contributor.authorKennedy, Jonathan
dc.contributor.authorDobson, Alan D. W.
dc.contributor.funderFederation of European Microbiological Societiesen
dc.contributor.funderScience Foundation Irelanden
dc.contributor.funderConsejo Nacional de Ciencia y Tecnologíaen
dc.date.accessioned2019-11-20T05:46:24Z
dc.date.available2019-11-20T05:46:24Z
dc.date.issued2019-06-14
dc.description.abstractCellulases 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.sponsorshipFederation of European Microbiological Societies (FEMSRG-2014-0007.R1); CONACyT-SEP (285816); Science Foundation Ireland (SSPC-2, 12/RC/2275)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleid1342en
dc.identifier.citationWierzbicka-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.2019.01342en
dc.identifier.doi10.3389/fmicb.2019.01342en
dc.identifier.eissn1664-302X
dc.identifier.endpage19en
dc.identifier.journaltitleFrontiers in Microbiologyen
dc.identifier.startpage1en
dc.identifier.urihttps://hdl.handle.net/10468/9133
dc.identifier.volume10en
dc.language.isoenen
dc.publisherFrontiers Mediaen
dc.relation.urihttps://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.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectCellulaseen
dc.subjectEndo-glucanaseen
dc.subjectGlycosyl hydrolase (GH)en
dc.subjectFunctional metagenomicsen
dc.subjectBiochemical characterisationen
dc.titleBiochemical characterization of a novel monospecific endo-β-1,4-glucanase belonging to GH Family 5 from a rhizosphere metagenomic libraryen
dc.typeArticle (peer-reviewed)en
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