In: Sydowia 74, (2022): 303-313; ISSN 0082-0598, DOI 10.12905/0380.sydowia74-2021-0303, Published online on January 21, 2022

Characterization and partial purification of
L-asparaginase produced by endophytic fungi from
cactus Cereus jamacaru

Marília Gomes da Silva Santos, Marília de Holanda Cavalcanti Maciel*, Minelli Albuquerque Sousa,
Aline Gleyce Julião Bomfim, Jadson Diogo Pereira Bezerra, Adalberto Pessoa Júnior &
Cristina Maria de Souza-Motta


Departamento de Micologia Prof. Chaves Batista, Universidade Federal de Pernambuco – UFPE. Rua Prof. Nelson Chaves,
s/n – Cidade Universitária. CEP: 50670-901. Recife, Pernambuco, Brazil
Setor de Micologia, Departamento de Biociências e Tecnologia, Instituto de Patologia Tropical e Saúde Pública (IPTSP),
Universidade Federal de Goiás (UFG), Rua 235, s/n, Setor Universitário, CEP: 74605-050. Goiânia, GO, Brazil
Departamento de Tecnologia Bioquímico-Farmacêutica, Universidade de São Paulo (USP), Av. Prof. Lineu Prestes, n. 580,
Bloco 16, Butantan, CEP: 05508000. São Paulo, SP, Brazil

* e-mail: macielmhc@gmail.com

Santos M.G.S., Maciel M.H.C., Sousa M.A., Bomfim A.G.J., Bezerra J.D.P., Júnior A.P. & Souza-Motta C.M. (2022) Characterisation
and partial purification of L-asparaginase produced by endophytic fungi from cactus Cereus jamacaru. – Sydowia 74:
303–313.

L-asparaginase (L-ASNase) is used in the treatment of acute lymphoblastic leukaemia and in food processing. This paper
evaluates the production, purification, and characterisation of L-ASNase from five endophytic fungi isolated from the cactus
Cereus jamacaru. All five filamentous fungi showed the ability to produce L-ASNase; however, Aspergillus flavus URM 6886
showed the highest enzymatic activity (0.12 U/ml). The L-ASNase produced by this fungus exhibited highest enzymatic activity
at pH 4.0 and 50 °C, and showed enzymatic activity over a wide range of pH values (4.0–9.2) at temperatures of 20 °C and 30 °C.
The activity of L-ASNase showed lower values in the presence of Co2+, Zn2+, Mn2+, and Ca2+ ions and was completely inactivated
in the presence of Mg2+ and Cu2+. Phenylmethanesulfonyl fluoride and iodoacetic acid caused significant increases of 122.9 % and
168.7 %, respectively, in enzyme activity. The optimal conditions for the purification of L-ASNase using a two-phase aqueous
system were 24.0 % (w/w) of 6000 g/mol PEG and 20.0 % (w/w) of citrate. Aspergillus flavus URM 6886 is indicated for future
studies to optimize the production of L-ASNase and its biotechnological applications, be it in the food or medicine industry.

Keywords: aqueous two-phase systems, asparagine, Aspergillus flavus, endophytic fungus, leukaemia.