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A new extracellular glutaminase and urease-free l-asparaginase from Meyerozyma guilliermondii

  • Biotechnology and Industrial Microbiology - Research Paper
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Abstract

l-Asparaginase (l-ASNase) is a potent chemotherapeutic drug employed to treat leukemia and lymphoma. Currently, l-ASNases for therapeutic use are obtained from Escherichia coli and Dickeya chrysanthemi (Erwinia chrysanthemi). Despite their therapeutic potential, enzymes from bacteria are subject to inducing immune responses, resulting in a higher number of side effects. Eukaryote producers, such as fungi, may provide therapeutic alternatives through enzymes that induce relatively less toxicity and immune responses. Additional expected benefits from yeast-derived enzymes include higher activity and stability in physiological conditions. This work describes the new potential therapeutic candidate l-ASNase from the yeast Meyerozyma guilliermondii. A statistical approach (full factorial central composite design) was used to optimize l-ASNase production, considering l-asparagine and glucose concentration, pH of the medium, and cultivation time as independent factors. In addition, the crude enzymes were biochemically characterized, in terms of temperature and optimal pH, thermostability, pH stability, and associated glutaminase or urease activities. Our results showed that enzyme production increased after supplementing a pH 4.0 medium with 1.0% l-asparagine and 0.5% glucose during 75 h of cultivation. Under these optimized conditions, l-ASNase production reached 26.01 U mL−1, which is suitable for scale-up studies. The produced l-ASNase exhibits maximal activity at 37 °C and pH 7.0 and is highly stable under physiological conditions. In addition, M. guilliermondii l-ASNase has no associated glutaminase or urease activities, demonstrating its potential as a promising antineoplastic agent.

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Funding

This study was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Grant No. 443953/2014–7) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES) (Finance Code 001).

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Authors and Affiliations

  1. Departamento de Ciências Biológicas, Universidade Estadual Do Centro-Oeste, Camargo Varela de Sá Street, 03, Guarapuava, Paraná State, 85040-080, Brazil

    Aline Ratuchne, Simone Cristine Izidoro, Susan Michelz Beitel & Adriana Knob

  2. Departamento de Biologia Geral E Aplicada, Universidade Estadual Paulista (UNESP), 24A Avenue, 1515, Rio Claro, São Paulo State, 13506-752, Brazil

    Lorena Tigre Lacerda

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  1. Aline Ratuchne
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  2. Simone Cristine Izidoro
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Correspondence to Adriana Knob.

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Ratuchne, A., Izidoro, S.C., Beitel, S.M. et al. A new extracellular glutaminase and urease-free l-asparaginase from Meyerozyma guilliermondii. Braz J Microbiol 54, 715–723 (2023). https://doi.org/10.1007/s42770-023-00939-x

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