Recombinantl-asparaginase production usingPichia pastoris(MUT(s)strain): establishment of conditions for growth and induction phases

Author(s):Show less -

Pillaca-Pullo, Omar ^{[1, 2]} ; Rodrigues, David ^[3] ; Sanchez-Moguel, Ignacio ^[1] ; Lopes, Andre ^[4] ; Pimenta, Marcela ^[1] ; Basi, Tajindar ^[5] ; Feitosa, Valker ^[1] ; Iris Zavaleta, Amparo ^[6] ; Monteiro, Gisele ^[1] ; Pessoa Jr, Adalberto ; Vitolo, Michele ^[7]

Affiliation

^[1] Univ Sao Paulo, Dept Tecnol Bioquim Farmaceut, Fac Ciencias Farmaceut, Sao Paulo - Brazil

^[2] Univ Privada San Juan Bautista, Escuela Med Humana, Lima - Peru

^[3] Univ Lisbon, Dept Bioengn, Inst Super Tecn, Lisbon - Portugal

^[4] Univ Estadual Campinas, Fac Pharmaceut Sci, Campinas - Brazil

^[5] Kings Coll London, Franklin Wilkins Bldg, London - England

^[6] Univ Nacl Mayor San Marcos, Mol Biol Lab, Fac Farm & Bioquim, Lima - Peru

^[7] Pessoa Jr, Jr., Adalberto, Univ Sao Paulo, Dept Tecnol Bioquim Farmaceut, Fac Ciencias Farmaceut, Sao Paulo - Brazil

• Document type: Journal article
• Source: JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY; v. 96, n. 1 AUG 2020.
• Web of Science Citations: 1

Abstract

BACKGROUND l-asparaginase (ASNase), a biopharmaceutical enzyme used in the treatment of childhood lymphoid malignancies, is commercially produced fromEscherichia coliandErwinia chrysanthemi.However, it causes severe adverse effects due to allergenic prokaryotic epitopes on the protein surface. ASNase II fromSaccharomyces cerevisiaecan be a promising alternative source of this enzyme. In this study, conditions to produce ASNase fromS. cerevisiaeexpressed inPichia pastorishave been investigated in shake flasks and 3L-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 degrees C and initial pH 6.0. CONCLUSION The experiments performed in shake flasks were scalable to a 3L-bioreactor, suggesting that this bioprocess could be scaled-up for industrial production. (c) 2020 Society of Chemical Industry (SCI) (AU)

• FAPESP's process: 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical
• Grantee: Adalberto Pessoa Junior
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/10789-1 - Combinatorial therapy using polymersomes decorated with transferrin and incorporated into chitosan hydrogels as smart drug delivery systems for melanoma tumor cells
• Grantee: André Moreni Lopes
• Support Opportunities: Research Grants - Young Investigators Grants