Development of a microbial platform for a new L-Asparaginase production

• Grant number: 21/01284-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2022
• End date: August 31, 2025
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Danielle Biscaro Pedrolli
• Grantee: Gabriela Barbosa de Paiva
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Biopharmaceuticals production has grown rapidly in the World. It's predicted the market for therapeutic enzymes will reach $ 6.3 billion by 2021. However, the production and purification process of these biomolecules is still costly. A solution to this problem may be the development of a new strain that generates high production yields and facilitates the extraction and purification process of the biomolecule by exporting it to the extracellular environment. In addition, a standardized platform may be useful for the production of a wide variety of biopharmaceuticals with reduced costs and individual development time. Within this context, this project proposes the development of a standardized platform for the production of biopharmaceuticals, using the bacterium Bacillus subtilis as chassis. As the therapeutic protein, the L-Asparaginase II from Aliivibrio fischeri will be produced. This enzyme has not yet been produced or characterized but has anti-leukemic potential based on its amino acid sequence. In this project, two release systems will be constructed. One of these is the L-Asparaginase II secretion mediated by a signal peptide and the other is the programmed lysis of the bacteria after consumption of the entire carbon source. These systems will be compared and the most efficient will be submitted to a scale up in bioreactors. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CRISTOVAO, RAQUEL O.; BARROS, RITA A. M.; MARRAMAQUE, TERESA P.; AGUIAR, GONCALO G.; ALMEIDA, MAFALDA R.; CARABINEIRO, SONIA A. C.; DE PAIVA, GABRIELA B.; PEDROLLI, DANIELLE B.; FREIRE, MARA G.; FARIA, JOAQUIM L.; et al. One-step purification of L-asparaginase from cell extracts using carbon xerogels. Separation and Purification Technology, v. 354, p. 9-pg., 2024-08-01. (21/06686-8, 18/06908-8, 21/01284-9)

NUNES, JOAO C. F.; ALMEIDA, MAFALDA R.; DE PAIVA, GABRIELA B.; PEDROLLI, DANIELLE B.; SANTOS-EBINUMA, VALERIA C.; NEVES, MARCIA C.; FREIRE, MARA G.; TAVARES, ANA P. M.. A flow-through strategy using supported ionic liquids for L-asparaginase purification. Separation and Purification Technology, v. 315, p. 11-pg., 2023-04-05. (21/01284-9)

BENTO, HEITOR B. S.; PAIVA, GABRIELA B.; ALMEIDA, MAFALDA R.; SILVA, CLAUDIA G.; CARVALHO, PEDRO J.; TAVARES, ANA P. M.; PEDROLLI, DANIELLE B.; SANTOS-EBINUMA, VALERIA C.. Aliivibrio fischeri L-Asparaginase production by engineered Bacillus subtilis: a potential new biopharmaceutical. Bioprocess and Biosystems Engineering, v. 45, n. 10, p. 10-pg., 2022-08-16. (18/06908-8, 20/15513-7, 21/01284-9, 21/06686-8)