One-step purification of L-asparaginase from cell extracts using carbon xerogels

• Tipo: Artigo em Revista Científica Internacional
• Data: 2024

Autores

Raquel O. Cristóvão (Autor) FEUP; R.A.M. Barros (Autor) Outra; Teresa P. Marramaque (Autor) Outra; Gonçalo G. Aguiar (Autor) Outra; Mafalda R. Almeida (Autor) Outra; Sónia A.C. Carabineiro (Autor) Outra; Gabriela B. de Paiva (Autor) Outra; Danielle B. Pedrolli (Autor) Outra; Mara G. Freire (Autor) Outra; Joaquim Luís Faria (Autor) FEUP; Valéria C. Santos-Ebinuma (Autor) Outra; Ana P.M. Tavares (Autor) Outra; Claudia G Silva (Autor) FEUP

Revista

Título: Separation and Purification Technology

Vol. 354 Nº 4

Página Inicial: 128969

ISSN: 1383-5866

Editora: Elsevier

Projetos Associados

LA LSRE-LCM - Financiamento Plurianual 2020-2023 para a Unidade LA LSRE-LCM Laboratório de Processos de Separação e Reacção - Laboratório de Catálise e Materiais

Outras Informações

ID Authenticus: P-011-3F7

DOI: 10.1016/j.seppur.2024.128969

Abstract (EN): L-asparaginase (ASNase, EC 3.5.1.1) is an enzyme with wide applications in the pharmaceutical sector and food processing industries. It is mainly used as a biotherapeutic for treating Acute Lymphoblastic Leukemia (ALL) and to reduce acrylamide formation in starchy compounds. Despite its relevance, current purification methods for microbial enzymes involve complex and expensive techniques. To overcome this drawback, Carbon Xerogels (CXs) were here investigated as novel adsorbents to be applied in an one-step ASNase purification process, using a flow-through-like setup, from a cell extract of genetically engineered Bacillus subtilis. Different operating conditions were studied for optimizing the adsorption onto CXs, including total protein concentration (3¿15 mg/mL), CXs amount (12, 18 and 24 mg), and adsorption volume of cell extract (1.5, 2.0 and 15 mL). Ultimately, CXs were packed into a column to evaluate the feasibility of semi-continuous ASNase purification. CXs have high affinity for other proteins present in the cell extract, while leaving ASNase in the supernatant or eluted sample. Purification folds of 2.5 and 3.8 for ASNase were obtained in batch and semi-continuous experiments, respectively, revealing the potential of CXs as novel adsorbent materials for ASNase purification directly from a complex matrix.

Idioma: Inglês

Tipo (Avaliação Docente): Científica