L-Asparaginase from Penicillium sizovae produced by a recombinant Komagataella phaffii strain
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| Main Author: | |
|---|---|
| Publication Date: | 2022 |
| Other Authors: | , , , , , , , , , |
| Format: | Article |
| Language: | eng |
| Source: | Repositório Institucional da UnB |
| Download full: | 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 |
Summary: | : 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. |
Internet
http://repositorio2.unb.br/jspui/handle/10482/48896https://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