L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.

Detalhes bibliográficos
Autor(a) principal: CARDOSO, S. L.
Data de Publicação: 2023
Outros Autores: SOUZA, P. M., RODRIGUES, K., MOTA, I. de S., FERREIRA FILHO, E., FAVARO, L. C. de L., ARAUJO, F. S., HOMEM-DE-MELLO, M., PESSOA, A., SILVEIRA, D., BAZZO, Y. M. F., MAGALHÃES, P. O.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
Texto Completo: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156949
https://doi.org/10.3390/pharmaceutics15092352
Resumo: The search for new drug-producing microorganisms is one of the most promising situations in current world scientific scenarios. The use of molecular biology as well as the cloning of protein and compound genes is already well established as the gold standard method of increasing productivity. Aiming at this increase in productivity, this work aims at the cloning, purification and in silico analysis of L-asparaginase from Fusarium proliferatum in Komagataella phaffii (Pichia pastoris) protein expression systems. The L-asparaginase gene (NCBI OQ439985) has been cloned into Pichia pastoris strains. Enzyme production was analyzed via the quantification of aspartic B-hydroxamate, followed by purification on a DEAE FF ion exchange column. The in silico analysis was proposed based on the combined use of various technological tools. The enzymatic activity found intracellularly was 2.84 IU/g. A purification factor of 1.18 was observed. The in silico analysis revealed the position of five important amino acid residues for enzymatic activity, and likewise, it was possible to predict a monomeric structure with a C-score of 1.59. The production of the enzyme L-asparaginase from F. proliferatum in P. pastoris was demonstrated in this work, being of great importance for the analysis of new methodologies in search of the production of important drugs in therapy.
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spelling L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.In silico analysisAsparaginaseHeterologous gene expressionFusarium proliferatumThe search for new drug-producing microorganisms is one of the most promising situations in current world scientific scenarios. The use of molecular biology as well as the cloning of protein and compound genes is already well established as the gold standard method of increasing productivity. Aiming at this increase in productivity, this work aims at the cloning, purification and in silico analysis of L-asparaginase from Fusarium proliferatum in Komagataella phaffii (Pichia pastoris) protein expression systems. The L-asparaginase gene (NCBI OQ439985) has been cloned into Pichia pastoris strains. Enzyme production was analyzed via the quantification of aspartic B-hydroxamate, followed by purification on a DEAE FF ion exchange column. The in silico analysis was proposed based on the combined use of various technological tools. The enzymatic activity found intracellularly was 2.84 IU/g. A purification factor of 1.18 was observed. The in silico analysis revealed the position of five important amino acid residues for enzymatic activity, and likewise, it was possible to predict a monomeric structure with a C-score of 1.59. The production of the enzyme L-asparaginase from F. proliferatum in P. pastoris was demonstrated in this work, being of great importance for the analysis of new methodologies in search of the production of important drugs in therapy.SAMUEL LEITE CARDOSO, UNIVERSITY OF BRASILIA; PAULA MONTEIRO SOUZA, UNIVERSITY OF BRASILIA; KELLY RODRIGUES, EMBRAPA AGROENERGIA; ISABELLA DE SOUZA MOTA, UNIVERSITY OF BRASILIA; EDIVALDO FERREIRA FILHO, UNIVERSITY OF BRASÍLIA; LEIA CECILIA DE LIMA FAVARO, CNPAE; FELIPE SALDANHA ARAUJO, UNIVERSITY OF BRASILIA; MAURICIO HOMEM-DE-MELLO, UNIVERSITY OF BRASILIA; ADALBERTO PESSOA, UNIVERSITY OF SÃO PAULO; DÂMARIS SILVEIRA, UNIVERSITY OF BRASILIA; YRIS MARIA FONSECA-BAZZO, UNIVERSITY OF BRASILIA; PÉROLA OLIVEIRA MAGALHÃES, UNIVERSITY OF BRASILIA.CARDOSO, S. L.SOUZA, P. M.RODRIGUES, K.MOTA, I. de S.FERREIRA FILHO, E.FAVARO, L. C. de L.ARAUJO, F. S.HOMEM-DE-MELLO, M.PESSOA, A.SILVEIRA, D.BAZZO, Y. M. F.MAGALHÃES, P. O.2023-09-27T20:24:15Z2023-09-27T20:24:15Z2023-09-272023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlePharmaceutics, v. 15, 2352, 2023.http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156949https://doi.org/10.3390/pharmaceutics15092352enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2023-09-27T20:24:15Zoai:www.alice.cnptia.embrapa.br:doc/1156949Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542023-09-27T20:24:15falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542023-09-27T20:24:15Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false
dc.title.none.fl_str_mv L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
title L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
spellingShingle L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
CARDOSO, S. L.
In silico analysis
Asparaginase
Heterologous gene expression
Fusarium proliferatum
title_short L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
title_full L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
title_fullStr L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
title_full_unstemmed L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
title_sort L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
author CARDOSO, S. L.
author_facet CARDOSO, S. L.
SOUZA, P. M.
RODRIGUES, K.
MOTA, I. de S.
FERREIRA FILHO, E.
FAVARO, L. C. de L.
ARAUJO, F. S.
HOMEM-DE-MELLO, M.
PESSOA, A.
SILVEIRA, D.
BAZZO, Y. M. F.
MAGALHÃES, P. O.
author_role author
author2 SOUZA, P. M.
RODRIGUES, K.
MOTA, I. de S.
FERREIRA FILHO, E.
FAVARO, L. C. de L.
ARAUJO, F. S.
HOMEM-DE-MELLO, M.
PESSOA, A.
SILVEIRA, D.
BAZZO, Y. M. F.
MAGALHÃES, P. O.
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv SAMUEL LEITE CARDOSO, UNIVERSITY OF BRASILIA; PAULA MONTEIRO SOUZA, UNIVERSITY OF BRASILIA; KELLY RODRIGUES, EMBRAPA AGROENERGIA; ISABELLA DE SOUZA MOTA, UNIVERSITY OF BRASILIA; EDIVALDO FERREIRA FILHO, UNIVERSITY OF BRASÍLIA; LEIA CECILIA DE LIMA FAVARO, CNPAE; FELIPE SALDANHA ARAUJO, UNIVERSITY OF BRASILIA; MAURICIO HOMEM-DE-MELLO, UNIVERSITY OF BRASILIA; ADALBERTO PESSOA, UNIVERSITY OF SÃO PAULO; DÂMARIS SILVEIRA, UNIVERSITY OF BRASILIA; YRIS MARIA FONSECA-BAZZO, UNIVERSITY OF BRASILIA; PÉROLA OLIVEIRA MAGALHÃES, UNIVERSITY OF BRASILIA.
dc.contributor.author.fl_str_mv CARDOSO, S. L.
SOUZA, P. M.
RODRIGUES, K.
MOTA, I. de S.
FERREIRA FILHO, E.
FAVARO, L. C. de L.
ARAUJO, F. S.
HOMEM-DE-MELLO, M.
PESSOA, A.
SILVEIRA, D.
BAZZO, Y. M. F.
MAGALHÃES, P. O.
dc.subject.por.fl_str_mv In silico analysis
Asparaginase
Heterologous gene expression
Fusarium proliferatum
topic In silico analysis
Asparaginase
Heterologous gene expression
Fusarium proliferatum
description The search for new drug-producing microorganisms is one of the most promising situations in current world scientific scenarios. The use of molecular biology as well as the cloning of protein and compound genes is already well established as the gold standard method of increasing productivity. Aiming at this increase in productivity, this work aims at the cloning, purification and in silico analysis of L-asparaginase from Fusarium proliferatum in Komagataella phaffii (Pichia pastoris) protein expression systems. The L-asparaginase gene (NCBI OQ439985) has been cloned into Pichia pastoris strains. Enzyme production was analyzed via the quantification of aspartic B-hydroxamate, followed by purification on a DEAE FF ion exchange column. The in silico analysis was proposed based on the combined use of various technological tools. The enzymatic activity found intracellularly was 2.84 IU/g. A purification factor of 1.18 was observed. The in silico analysis revealed the position of five important amino acid residues for enzymatic activity, and likewise, it was possible to predict a monomeric structure with a C-score of 1.59. The production of the enzyme L-asparaginase from F. proliferatum in P. pastoris was demonstrated in this work, being of great importance for the analysis of new methodologies in search of the production of important drugs in therapy.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-27T20:24:15Z
2023-09-27T20:24:15Z
2023-09-27
2023
dc.type.driver.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv Pharmaceutics, v. 15, 2352, 2023.
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156949
https://doi.org/10.3390/pharmaceutics15092352
identifier_str_mv Pharmaceutics, v. 15, 2352, 2023.
url http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156949
https://doi.org/10.3390/pharmaceutics15092352
dc.language.iso.fl_str_mv eng
language eng
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