Título:	L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis.
Autoria:	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.
Afiliação:	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.
Ano de publicação:	2023
Referência:	Pharmaceutics, v. 15, 2352, 2023.
Conteúdo:	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.
NAL Thesaurus:	Asparaginase Heterologous gene expression Fusarium proliferatum
Palavras-chave:	In silico analysis
Digital Object Identifier:	https://doi.org/10.3390/pharmaceutics15092352
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CNPAE)