Recombinant L-asparaginase production using Pichia pastoris (MUTs strain): establishment of conditions for growth and induction phases
| Autor: | André Moreni Lopes, Tajindar Basi, Ignacio Sánchez-Moguel, Omar Pillaca-Pullo, Marcela Pimenta, Valker Araujo Feitosa, Michele Vitolo, Amparo I. Zavaleta, Gisele Monteiro, Adalberto Pessoa, David Rodrigues |
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| Rok vydání: | 2021 |
| Předmět: |
General Chemical Engineering
Saccharomyces cerevisiae 02 engineering and technology 010501 environmental sciences medicine.disease_cause 01 natural sciences Pichia pastoris law.invention Inorganic Chemistry chemistry.chemical_compound law medicine Glycerol Inducer Bioprocess Waste Management and Disposal Escherichia coli 0105 earth and related environmental sciences chemistry.chemical_classification biology Renewable Energy Sustainability and the Environment Chemistry Organic Chemistry equipment and supplies 021001 nanoscience & nanotechnology biology.organism_classification Pollution Fuel Technology Enzyme Biochemistry Recombinant DNA 0210 nano-technology Biotechnology SACCHAROMYCES |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| Popis: | BACKGROUND: l‐asparaginase (ASNase), a biopharmaceutical enzyme used in the treatment of childhood lymphoid malignancies, is commercially produced from Escherichia coli and Erwinia chrysanthemi. However, it causes severe adverse effects due to allergenic prokaryotic epitopes on the protein surface. ASNase II from Saccharomyces cerevisiae can be a promising alternative source of this enzyme. In this study, conditions to produce ASNase from S. cerevisiae expressed in Pichia pastoris have been investigated in shake flasks and 3 L‐bioreactor. We evaluated if medium composition, concentration of carbon source (i.e. glycerol), growth time, concentration of inducer (i.e. methanol), temperature and initial pH influenced both biomass and ASNase expression. RESULTS: Biomass of around 53 g L–¹ and ASNase volumetric activity of 710 U L–¹ were achieved using the buffered glycerol‐complex medium (BMGY) containing 40 g L–¹ glycerol, with induction after 141 h using 3.0% (v/v) methanol, at 20 °C and initial pH 6.0. CONCLUSION: The experiments performed in shake flasks were scalable to a 3 L‐bioreactor, suggesting that this bioprocess could be scaled‐up for industrial production. © 2020 Society of Chemical Industry (SCI) |
| Databáze: | OpenAIRE |
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