Gomes José Gabriel da Silva, Brandão Larisse Cadeira, Pinheiro Daniel Pascoalino, Pontes Larissa Queiroz, Carneiro Rômulo Farias, Quintela Bárbara Cibelle Soares Farias, Marinho Anna Carolina Machado, Furtado Gilvan Pessoa, Rocha Bruno Anderson Matias
Department of Biochemistry and Molecular Biology, Federal University of Ceara, Fortaleza, Brazil.
Departament of Fishing Engineering, Federal University of Ceara, Fortaleza, Brazil; Oswaldo Cruz Foundation - Fiocruz Ceara, Eusebio, Ceara, Brazil.
Int J Biol Macromol. 2024 Aug;275(Pt 2):133731. doi: 10.1016/j.ijbiomac.2024.133731. Epub 2024 Jul 8.
l-asparaginases play a crucial role in the treatment of acute lymphoblastic leukemia (ALL), a type of cancer that mostly affects children and teenagers. However, it is common for these molecules to cause adverse reactions during treatment. These downsides ignite the search for novel asparaginases to mitigate these problems. Thus, this work aimed to produce and characterize a recombinant asparaginase from Phaseolus vulgaris (Asp-P). In this study, Asp-P was expressed in Escherichia coli with high yields and optimum activity at 40 °C, pH 9.0. The enzyme K and V values were 7.05 mM and 1027 U/mg, respectively. Asp-P is specific for l-asparagine, showing no activity against l-glutamine and other amino acids. The enzyme showed a higher cytotoxic effect against Raji than K562 cell lines, but only at high concentrations. In silico analysis indicated that Asp-P has lower immunogenicity than a commercial enzyme. Asp-P induced biofilm formation by Candida sp. due to sublethal dose, showing an underexplored potential of asparaginases. The absence of glutaminase activity, lower immunogenicity and optimal activity similar to physiological temperature conditions are characteristics that indicate Asp-P as a potential new commercial enzyme in the treatment of ALL and its underexplored application in the treatment of other diseases.
L-天冬酰胺酶在急性淋巴细胞白血病（ALL）的治疗中起着关键作用，ALL是一种主要影响儿童和青少年的癌症。然而，这些分子在治疗过程中引起不良反应是很常见的。这些缺点引发了对新型天冬酰胺酶的探索，以减轻这些问题。因此，这项工作旨在生产并表征来自菜豆的重组天冬酰胺酶（Asp-P）。在本研究中，Asp-P在大肠杆菌中高效表达，在40°C、pH 9.0时具有最佳活性。该酶的K值和V值分别为7.05 mM和1027 U/mg。Asp-P对L-天冬酰胺具有特异性，对L-谷氨酰胺和其他氨基酸无活性。该酶对Raji细胞系的细胞毒性作用高于K562细胞系，但仅在高浓度时如此。计算机分析表明，Asp-P的免疫原性低于一种商业酶。Asp-P由于亚致死剂量诱导念珠菌属形成生物膜，显示出天冬酰胺酶尚未被充分探索的潜力。缺乏谷氨酰胺酶活性、较低的免疫原性以及与生理温度条件相似的最佳活性，这些特征表明Asp-P是治疗ALL的一种潜在新型商业酶，并且在治疗其他疾病方面具有尚未被充分探索的应用。
