Generation of a library of carbohydrate-active enzymes for plant biomass deconstruction
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , |
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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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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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:RCAAP2024-11-20T18:36:17Zoai:repositorio.ul.pt:10400.5/24345Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-11-20T18:36:17Repositório 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 |
mluisa.alvim@gmail.com |
_version_ |
1817549321420144640 |