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Title: L-Asparaginase type II from fusarium proliferatum : heterologous expression and in silico analysis
Authors: Cardoso, Samuel Leite
Souza, Paula Monteiro de
Rodrigues, Kelly Barreto
Mota, Isabella de Souza
Ferreira Filho, Edivaldo Ximenes
Fávaro, Léia Cecilia de Lima
Araujo, Felipe Saldanha de
Homem de Mello, Mauricio
Pessoa, Adalberto
Duarte, Dâmaris Silveira
Bazzo, Yris Maria Fonseca
Batista, Pérola de Oliveira Magalhães Dias
metadata.dc.identifier.orcid: https://orcid.org/0000-0003-1824-2211
https://orcid.org/0009-0002-9014-3488
https://orcid.org/0000-0003-4947-8753
https://orcid.org/0000-0003-3260-0310
https://orcid.org/0000-0002-4541-9177
https://orcid.org/0000-0003-3260-0310
https://orcid.org/0000-0002-4541-9177
https://orcid.org/0000-0002-5268-8690
https://orcid.org/0000-0003-1851-5224
metadata.dc.contributor.affiliation: University of Brasilia, Health Science School
University of Brasilia, Health Science School
Brazilian Agricultural Research Corporation—EMBRAPA Agroenergia
University of Brasilia, Health Science School
University of Brasilia, Institute of Biological Sciences
Brazilian Agricultural Research Corporation—EMBRAPA Agroenergia
University of Brasilia, Health Science School
University of São Paulo, Department of Biochemical and Pharmaceutical Technology
University of Brasilia, Health Science School
University of Brasilia, Health Science School
University of Brasilia, Health Science School
University of Brasilia, Health Science School
Assunto:: L -asparaginase
Silos graneleiros - análise
Fungos fitopatogênicos
Issue Date: 20-Sep-2023
Publisher: MDPI
Citation: CARDOSO, Samuel Leite et al. L-Asparaginase type II from fusarium proliferatum: heterologous expression and in silico analysis. Pharmaceutics, [S. l.], v. 15, n. 9, 2352, 2023. DOI: https://doi.org/10.3390/pharmaceutics15092352. Disponível em: https://www.mdpi.com/1999-4923/15/9/2352. Acesso em: 20 set. 2024.
Abstract: 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.
metadata.dc.description.unidade: Faculdade de Ciências da Saúde (FS)
Instituto de Ciências Biológicas (IB)
Licença:: Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).
DOI: https://doi.org/10.3390/pharmaceutics15092352
Appears in Collections:Artigos publicados em periódicos e afins

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