Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability

Detalhes bibliográficos
Autor(a) principal: Ghosh, Arabinda
Data de Publicação: 2013
Outros Autores: Luís, Ana Sofia, Brás, Joana L. A., Pathaw, Neeta, Chrungoo, Nikhil K., Fontes, Carlos M. G. A., Goyal, Arun
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.5/7208
Resumo: Articles in International Journals
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spelling Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stabilityAdsorptionBinding analysisCarbohydratesClostridiumFluorescenceHydrodynamicsMeltingPolysaccharidesArticles in International JournalsThis study investigated the role of CBM35 from Clostridium thermocellum (CtCBM35) in polysaccharide recognition. CtCBM35 was cloned into pET28a (+) vector with an engineered His6 tag and expressed in Escherichia coli BL21 (DE3) cells. A homogenous 15 kDa protein was purified by immobilized metal ion chromatography (IMAC). Ligand binding analysis of CtCBM35 was carried out by affinity electrophoresis using various soluble ligands. CtCBM35 showed a manno-configured ligand specific binding displaying significant association with konjac glucomannan (Ka = 14.3×104 M−1), carob galactomannan (Ka = 12.4×104 M−1) and negligible association (Ka = 12 µM−1) with insoluble mannan. Binding of CtCBM35 with polysaccharides which was calcium dependent exhibited two fold higher association in presence of 10 mM Ca2+ ion with konjac glucomannan (Ka = 41×104 M−1) and carob galactomannan (Ka = 30×104 M−1). The polysaccharide binding was further investigated by fluorescence spectrophotometric studies. On binding with carob galactomannan and konjac glucomannan the conformation of CtCBM35 changed significantly with regular 21 nm peak shifts towards lower quantum yield. The degree of association (Ka) with konjac glucomannan and carob galactomannan, 14.3×104 M−1 and 11.4×104 M−1, respectively, corroborated the findings from affinity electrophoresis. The association of CtCBM35with konjac glucomannan led to higher free energy of binding (ΔG) −25 kJ mole−1 as compared to carob galactomannan (ΔG) −22 kJ mole−1. On binding CtCBM35 with konjac glucomannan and carob galactomannan the hydrodynamic radius (RH) as analysed by dynamic light scattering (DLS) study, increased to 8 nm and 6 nm, respectively, from 4.25 nm in absence of ligand. The presence of 10 mM Ca2+ ions imparted stiffer orientation of CtCBM35 particles with increased RH of 4.52 nm. Due to such stiffer orientation CtCBM35 became more thermostable and its melting temperature was shifted to 70°C from initial 50°C.Michael Massiah, George Washington University, United States of AmericaRepositório da Universidade de LisboaGhosh, ArabindaLuís, Ana SofiaBrás, Joana L. A.Pathaw, NeetaChrungoo, Nikhil K.Fontes, Carlos M. G. A.Goyal, Arun2014-09-25T15:32:57Z2013-12-062013-12-06T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.5/7208engGhosh, A., et al. (2013). Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability. PLoS ONE, 8(12): e80415, 1-11. doi: 10.1371/ journal.pone.00804151932-6203 (online)10.1371/journal.pone.0080415info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-06T14:37:57Zoai:www.repository.utl.pt:10400.5/7208Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T16:54:25.967644Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
title Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
spellingShingle Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
Ghosh, Arabinda
Adsorption
Binding analysis
Carbohydrates
Clostridium
Fluorescence
Hydrodynamics
Melting
Polysaccharides
title_short Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
title_full Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
title_fullStr Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
title_full_unstemmed Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
title_sort Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability
author Ghosh, Arabinda
author_facet Ghosh, Arabinda
Luís, Ana Sofia
Brás, Joana L. A.
Pathaw, Neeta
Chrungoo, Nikhil K.
Fontes, Carlos M. G. A.
Goyal, Arun
author_role author
author2 Luís, Ana Sofia
Brás, Joana L. A.
Pathaw, Neeta
Chrungoo, Nikhil K.
Fontes, Carlos M. G. A.
Goyal, Arun
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Ghosh, Arabinda
Luís, Ana Sofia
Brás, Joana L. A.
Pathaw, Neeta
Chrungoo, Nikhil K.
Fontes, Carlos M. G. A.
Goyal, Arun
dc.subject.por.fl_str_mv Adsorption
Binding analysis
Carbohydrates
Clostridium
Fluorescence
Hydrodynamics
Melting
Polysaccharides
topic Adsorption
Binding analysis
Carbohydrates
Clostridium
Fluorescence
Hydrodynamics
Melting
Polysaccharides
description Articles in International Journals
publishDate 2013
dc.date.none.fl_str_mv 2013-12-06
2013-12-06T00:00:00Z
2014-09-25T15:32:57Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.5/7208
url http://hdl.handle.net/10400.5/7208
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Ghosh, A., et al. (2013). Deciphering ligand specificity of a Clostridium thermocellum family 35 carbohydrate binding module (CtCBM35) for Gluco- and Galacto- Substituted mannans and Its calcium induced stability. PLoS ONE, 8(12): e80415, 1-11. doi: 10.1371/ journal.pone.0080415
1932-6203 (online)
10.1371/journal.pone.0080415
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Michael Massiah, George Washington University, United States of America
publisher.none.fl_str_mv Michael Massiah, George Washington University, United States of America
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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