Please use this identifier to cite or link to this item: http://hdl.handle.net/10553/51925
DC FieldValueLanguage
dc.contributor.authorZafra, Olga
dc.contributor.authorFraile, Sofía
dc.contributor.authorGutierrez, Carlos
dc.contributor.authorHaro, Amparo
dc.contributor.authorPáez-Espino, A. David
dc.contributor.authorJiménez, Jose I.
dc.contributor.authorDe Lorenzo, Víctor
dc.contributor.otherZafra, Olga
dc.contributor.otherGutierrez, Carlos
dc.contributor.otherJimenez, Jose
dc.contributor.otherde Lorenzo, Victor
dc.date.accessioned2018-11-25T05:28:27Z-
dc.date.available2018-11-25T05:28:27Z-
dc.date.issued2011
dc.identifier.issn1462-2912
dc.identifier.urihttp://hdl.handle.net/10553/51925-
dc.description.abstractFunctional studies of biodegradative activities in environmental microorganisms require molecular tools for monitoring catabolic enzymes in the members of the native microbiota. To this end, we have generated repertories of single-domain VHH fragments of camel immunoglobulins (nanobodies) able to interact with multiple proteins that are descriptors of environmentally relevant processes. For this, we immunized Camelus dromedarius with a cocktail of up to 12 purified enzymes that are representative of major types of detoxifying activities found in aerobic and anaerobic microorganisms. Following the capture of the antigen-binding modules from the mRNA of the camel lymphocytes and the selection of sub-libraries for each of the enzymes in a phage display system we found a large number of VHH modules that interacted with each of the antigens. Those associated to the enzyme 2,3 dihydroxybiphenyl dioxygenase of Burkholderia xenovorans LB400 (BphC) and the arsenate reductase of Staphylococcus aureus (ArsC) were examined in detail and found to hold different qualities that were optimal for distinct protein recognition procedures. The repertory of anti-BphC VHHs included variants with a strong affinity and specificity for linear epitopes of the enzyme. When the anti-BphC VHH library was recloned in a prokaryotic intracellular expression system, some nanobodies were found to inhibit the dioxygenase activity in vivo. Furthermore, anti-ArsC VHHs were able to discriminate between proteins stemming from different enzyme families. The easiness of generating large collections of binders with different properties widens considerably the molecular toolbox for analysis of biodegradative bacteria and opens fresh possibilities of monitoring protein markers and activities in the environment. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.
dc.publisher1462-2912
dc.relation.ispartofEnvironmental Microbiology
dc.sourceEnvironmental Microbiology[ISSN 1462-2912],v. 13 (4), p. 960-974
dc.titleMonitoring biodegradative enzymes with nanobodies raised in Camelus dromedarius with mixtures of catabolic proteins
dc.typeinfo:eu-repo/semantics/Articlees
dc.typeArticlees
dc.identifier.doi10.1111/j.1462-2920.2010.02401.x
dc.identifier.scopus79951933331
dc.identifier.isi000289116700011
dcterms.isPartOfEnvironmental Microbiology
dcterms.sourceEnvironmental Microbiology[ISSN 1462-2912],v. 13 (4), p. 960-974
dc.contributor.authorscopusid6507568708
dc.contributor.authorscopusid6508063810
dc.contributor.authorscopusid7202545218
dc.contributor.authorscopusid57213261824
dc.contributor.authorscopusid57198222052
dc.contributor.authorscopusid46761391400
dc.contributor.authorscopusid15922896500
dc.contributor.authorscopusid7005588312
dc.description.lastpage974
dc.description.firstpage960
dc.relation.volume13
dc.type2Artículoes
dc.identifier.wosWOS:000289116700011
dc.contributor.daisngid2540831
dc.contributor.daisngid3004896
dc.contributor.daisngid475608
dc.contributor.daisngid7513671
dc.contributor.daisngid7010607
dc.contributor.daisngid1243081
dc.contributor.daisngid50093
dc.identifier.investigatorRIDK-8110-2014
dc.identifier.investigatorRIDE-2989-2012
dc.identifier.investigatorRIDNo ID
dc.identifier.investigatorRIDNo ID
dc.date.coverdateAbril 2011
dc.identifier.ulpgces
dc.description.sjr3,33
dc.description.jcr5,843
dc.description.sjrqQ1
dc.description.jcrqQ1
dc.description.scieSCIE
item.fulltextSin texto completo-
item.grantfulltextnone-
crisitem.author.deptGIR IUIBS: Medicina Veterinaria e Investigación Terapéutica-
crisitem.author.deptIU de Investigaciones Biomédicas y Sanitarias-
crisitem.author.orcid0000-0003-0764-7408-
crisitem.author.parentorgIU de Investigaciones Biomédicas y Sanitarias-
crisitem.author.fullNameGutiérrez Cabrera,Carlos Javier-
Appears in Collections:Artículos
Show simple item record

SCOPUSTM   
Citations

26
checked on Mar 24, 2024

WEB OF SCIENCETM
Citations

18
checked on Oct 2, 2022

Page view(s)

26
checked on Jan 20, 2024

Google ScholarTM

Check

Altmetric


Share



Export metadata



Items in accedaCRIS are protected by copyright, with all rights reserved, unless otherwise indicated.