L-Asparaginase from Penicillium sizovae produced by a recombinant Komagataella phaffii strain

• Autor(a) principal: Freitas, Marcela Medeiros de
• Data de Publicação: 2022
• Outros Autores: Souza, Paula Monteiro de, Homem de Mello, Mauricio, Fonseca-Bazzo, Yris Maria, Silveira, Dâmaris, Ferreira Filho, Edivaldo Ximenes, Pessoa Junior, Adalberto, Sarker, Dipak, Timson, David, Inácio, João, Magalhães, Pérola de Oliveira
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UnB
• Texto Completo: http://repositorio2.unb.br/jspui/handle/10482/48896; https://doi.org/10.3390/ph15060746; https://orcid.org/0000-0002-6112-9903; https://orcid.org/0000-0003-1824-2211; https://orcid.org/0000-0002-4541-9177; https://orcid.org/0000-0002-1230-3207; https://orcid.org/0000-0003-1851-5224; https://orcid.org/0000-0003-4947-8753; https://orcid.org/0000-0002-5268-8690; https://orcid.org/0000-0002-2912-415X; https://orcid.org/0000-0002-0985-8818; https://orcid.org/0000-0001-8011-6940
• topic: L-Asparaginase; Leucemia linfoblástica aguda (LLA); Clonagem; Fungos

: L-asparaginase is an important enzyme in the pharmaceutical field used as treatment for acute lymphoblastic leukemia due to its ability to hydrolyze L-asparagine, an essential amino acid synthesized by normal cells, but not by neoplastic cells. Adverse effects of L-asparaginase formulations are associated with its glutaminase activity and bacterial origin; therefore, it is important to find new sources of L-asparaginase produced by eukaryotic microorganisms with low glutaminase activity. This work aimed to identify the L-asparaginase gene sequence from Penicillium sizovae, a filamentous fungus isolated from the Brazilian Savanna (Cerrado) soil with low glutaminase activity, and to biosynthesize higher yields of this enzyme in the yeast Komagataella phaffii. The L-asparaginase gene sequence of P. sizovae was identified by homology to L-asparaginases from species of Penicillium of the section Citrina: P. citrinum and P. steckii. Partial L-asparaginase from P. sizovae, lacking the periplasmic signaling sequence, was cloned, and expressed intracellularly with highest enzymatic activity achieved by a MUT+ clone cultured in BMM expression medium; a value 5-fold greater than that obtained by native L-asparaginase in P. sizovae cells. To the best of our knowledge, this is the first literature report of the heterologous production of an L-asparaginase from a filamentous fungus by a yeast.