Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction

Detalhes bibliográficos
Autor(a) principal: Cardoso, Vânia
Data de Publicação: 2022
Outros Autores: Bras, Joana L. A., Costa, Ines F., Ferreira, Luis M. A., Gama, Luis, Vincentelli, Renaud, Henrissat, Bernard, Fontes, Carlos M.G.A.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.5/24345
Resumo: Áreas de pesquisa: Biochemistry & Molecular Biology ; Chemistry
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spelling Generation of a library of carbohydrate-active enzymes for plant biomass deconstructionCarbohydrate-active enzymes (CAZymes)Plant biomassGene synthesisPCRHigh-throughput (HTP) cloningHTP expressionÁreas de pesquisa: Biochemistry & Molecular Biology ; ChemistryIn nature, the deconstruction of plant carbohydrates is carried out by carbohydrate-active enzymes (CAZymes). A high-throughput (HTP) strategy was used to isolate and clone 1476 genes obtained from a diverse library of recombinant CAZymes covering a variety of sequence-based families, enzyme classes, and source organisms. All genes were successfully isolated by either PCR (61%) or gene synthesis (GS) (39%) and were subsequently cloned into Escherichia coli expression vectors. Most proteins (79%) were obtained at a good yield during recombinant expression. A significantly lower number (p < 0.01) of proteins from eukaryotic (57.7%) and archaeal (53.3%) origin were soluble compared to bacteria (79.7%). Genes obtained by GS gave a significantly lower number (p = 0.04) of soluble proteins while the green fluorescent protein tag improved protein solubility (p = 0.05). Finally, a relationship between the amino acid composition and protein solubility was observed. Thus, a lower percentage of non-polar and higher percentage of negatively charged amino acids in a protein may be a good predictor for higher protein solubility in E. coli. The HTP approach presented here is a powerful tool for producing recombinant CAZymes that can be used for future studies of plant cell wall degradation. Successful production and expression of soluble recombinant proteins at a high rate opens new possibilities for the high-throughput production of targets from limitless sourcesMDPIRepositório da Universidade de LisboaCardoso, VâniaBras, Joana L. A.Costa, Ines F.Ferreira, Luis M. A.Gama, LuisVincentelli, RenaudHenrissat, BernardFontes, Carlos M.G.A.2022-05-22T22:38:43Z2022-04-052022-04-05T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.5/24345engCardoso V, Brás JLA, Costa IF, Ferreira LMA, Gama LT, Vincentelli R, Henrissat B, Fontes CMGA. 2022. Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction. Internal Journal of Molecular Sciences, 23(7):4024. DOI 10.3390/ ijms2307402410.3390/ ijms230740241422-0067info: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:RCAAP2023-03-06T14:54:00Zoai:www.repository.utl.pt:10400.5/24345Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T17:08:25.109843Repositório 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 Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
title Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
spellingShingle Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
Cardoso, Vânia
Carbohydrate-active enzymes (CAZymes)
Plant biomass
Gene synthesis
PCR
High-throughput (HTP) cloning
HTP expression
title_short Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
title_full Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
title_fullStr Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
title_full_unstemmed Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
title_sort Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
author Cardoso, Vânia
author_facet Cardoso, Vânia
Bras, Joana L. A.
Costa, Ines F.
Ferreira, Luis M. A.
Gama, Luis
Vincentelli, Renaud
Henrissat, Bernard
Fontes, Carlos M.G.A.
author_role author
author2 Bras, Joana L. A.
Costa, Ines F.
Ferreira, Luis M. A.
Gama, Luis
Vincentelli, Renaud
Henrissat, Bernard
Fontes, Carlos M.G.A.
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Cardoso, Vânia
Bras, Joana L. A.
Costa, Ines F.
Ferreira, Luis M. A.
Gama, Luis
Vincentelli, Renaud
Henrissat, Bernard
Fontes, Carlos M.G.A.
dc.subject.por.fl_str_mv Carbohydrate-active enzymes (CAZymes)
Plant biomass
Gene synthesis
PCR
High-throughput (HTP) cloning
HTP expression
topic Carbohydrate-active enzymes (CAZymes)
Plant biomass
Gene synthesis
PCR
High-throughput (HTP) cloning
HTP expression
description Áreas de pesquisa: Biochemistry & Molecular Biology ; Chemistry
publishDate 2022
dc.date.none.fl_str_mv 2022-05-22T22:38:43Z
2022-04-05
2022-04-05T00:00:00Z
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/10400.5/24345
url http://hdl.handle.net/10400.5/24345
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Cardoso V, Brás JLA, Costa IF, Ferreira LMA, Gama LT, Vincentelli R, Henrissat B, Fontes CMGA. 2022. Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction. Internal Journal of Molecular Sciences, 23(7):4024. DOI 10.3390/ ijms23074024
10.3390/ ijms23074024
1422-0067
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 MDPI
publisher.none.fl_str_mv MDPI
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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