Filamentous fungi producing l-asparaginase with low glutaminase activity isolated from Brazilian savanna soil

Marcela Freitas, Paula Souza, Samuel Cardoso, Kellen Cruvinel, Letícia Santos Abrunhosa, Edivaldo X. Ferreira Filho, João Inácio, Danilo Batista Pinho, Adalberto Pessoa, Pérola O. Magalhães

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L-asparaginase is an enzyme used as treatment for acute lymphoblastic leukemia (ALL) due to its ability to hydrolyze L-asparagine, an essential amino acid synthesized by normal cells unlike neo-plastic cells. The 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-producing eukaryotic microorganisms with low glutaminase activity. This work evaluated the biotechnological potential of filamentous fungi isolated from Brazilian Savanna soil and plants for L-asparaginase production. Thirty-nine isolates were screened for enzyme production using the plate assay, followed by measuring enzymatic activity in cells after submerged fermentation. The variables influencing L-asparaginase production were evaluated using Plackett–Burman design. Cell disruption methods were evaluated for L-asparaginase release. Penicillium sizovae 2DSST1 and Fusarium proliferatum DCFS10 showed the highest L-asparaginase activity levels and the lowest glutaminase activity levels. Penicillium sizovae L-asparaginase was repressed by carbon sources, whereas higher carbon concentrations enhanced L-asparaginase by F. proliferatum. Maximum enzyme productivity, specific enzyme yield and the biomass conversion factor in the enzyme increased after Plackett–Burman design. Freeze-grinding released 5-fold more L-asparaginase from cells than sonication. This study shows two species, which have not yet been reported, as sources of L-asparaginase with possible reduced immunogenicity for ALL therapy.

Original languageEnglish
Article number1268
Issue number8
Publication statusPublished - 17 Aug 2021

Bibliographical note

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© 2021 by the authors. Licensee MDPI, Basel, Switzerland.


  • Acute lymphoblastic leukemia
  • Filamentous fungi
  • Fusarium fujikuroi species complex
  • L-asparaginase
  • Penicillium
  • Plackett–Burman design


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