Título:	L-Asparaginase type II from fusarium proliferatum : heterologous expression and in silico analysis
Autor(es):	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
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
Afiliação do autor:	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
Data de publicação:	20-Set-2023
Editora:	MDPI
Referência:	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.
Unidade Acadêmica:	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
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