Sequencing and characterization of an L-asparaginase gene from a new species of Penicillium section Citrina isolated from Cerrado

Abstract

The enzyme L-asparaginase (L-ASNase) is used in the treatment of Acute Lymphoblastic Leukemia. The preparations of this enzyme for clinical use are derived from bacterial sources and its use is associated with serious adverse reactions. In this context, it is important to find new sources of L-ASNase. In this work, the Placket-Burman Experimental Design (PBD) was used to determine the influence of the variables on the L-ASNase production then it was followed by a 28–4 Factorial Fractional Design (FFD). The results obtained from PBD have shown a range of L-ASNase activity, from 0.47 to 1.77 U/gcell and the results obtained from FFD have showed a range of L-ASNase activity, from 1.10 to 2.36 U/gcell. L-proline and ammonium sulfate were identified as of significant positive variables on this production enzyme by Penicillium cerradense sp. nov. The precise identification of this new species was confirmed by morphological characteristics and sequence comparisons of the nuclear 18S-5.8S-28S partial nrDNA including the ITS1 and ITS2 regions, RNA polymerase II, β-tubulin and calmodulin genomic regions. The genetic sequence coding for the L-ASNase was obtained after carrying out a full genome sequencing. The L-ASNase expressed by P. cerradense sp. nov may have promising antineoplastic properties.

L-Asparaginase from Penicillium sizovae Produced by a Recombinant Komagataella phaffii Strain

“…The phylogenetic proximity of these Penicillium species that belong to section Citrina led to the identification of L-asparaginase from P. sizovae, a filamentous fungus isolated from the soil of the global biodiversity hotspot Cerrado in Brazil. This sequence showed homology to another L-asparaginase gene from a novel P. cerradense, a filamentous fungus isolated from the Brazilian Savanna, recently identified by this research group [45].…”

Desirable L-asparaginases for treating cancer and current research trends

“…L-asparaginases must have a micromolar K m for asparagine in order to remove it from the blood ( Rigouin et al, 2017 ). To get rid of the circulating L-asparagine in ALL patients, type II L-asparaginases from E. coli or E. chrysanthemi are utilized ( Radadiya et al, 2020 ; Andrade et al, 2021 ). Wild type II L-asparaginases exhibit high selectivity for L-asparagine and low glutaminase side activity (2%–10%).…”

Morphological and phylogenetic analyses reveal two new Penicillium species isolated from the ancient Great Wall loess in Beijing, China

“…Furthermore, members of this section have a high potential to produce the mycotoxins citrinin (Houbraken et al, 2011b;Dutra-Silva et al, 2021;Yin et al, 2021). Currently, this section includes 47 species (Houbraken et al, 2020;Andrade et al, 2021;Nguyen et al, 2021;Ashtekar et al, 2022;Tan and Shivas, 2022;Visagie and Yilmaz, 2023). Penicillium sect.…”