Scale up of L-asparaginase production employing genetically modified microorganism

• Grant number: 20/15513-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: January 01, 2021
• End date: May 31, 2022
• Field of knowledge: Engineering - Chemical Engineering
• Agreement: Fundação para a Ciência e a Tecnologia (FCT)
• Principal Investigator: Valéria de Carvalho Santos Ebinuma
• Grantee: Heitor Buzetti Simões Bento
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated research grant: 18/06908-8 - Development of sustainable nanomaterials for the purification of antileukemic drugs, AP.R

Abstract

In the health area, this project has a strong component of newbiotechnologies and advanced solutions for the production of productsbiopharmaceuticals to be used as therapeutic agents in various typesof cancer, specifically, in leukemia. L-asparaginase (E.C.3.5.1.1)(ASNase) is the enzyme that catalyzes the hydrolysis reaction of the amino group ofside chain of the amino acid L-asparagine, releasing aspartic acid and ammonia,being widely used as an anti-neoplastic agent. This enzyme can beproduced by a series of microorganisms, however, there is a need fornew producing sources with higher yields and less adverse effects.Thus, the objective of this project is to evaluate the production of L-asparaginase by astrain of Bacillus subtilis that produces Aliivibrio fischeri L-Asparaginase IIrotary shaker and scale up the process for tank bioreactoragitated. Initially, a study will be carried out on a rotary shaker evaluating,induction time, inductor concentration, inoculum size andcultivation. In parallel, cell disruption tests will be carried out toimprove extraction of the intracellular enzyme. Subsequently, scale up will be doneprocess for agitated tank bioreactor and some parameters will be evaluatedhydrodynamics related to the production process. The development ofnew L-asparaginase production lines and production process,will allow to obtain a therapy with higher yields, more effective and withreduced side effects.

Scientific publications

(10)

(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)

DE CASTRO, TARSILA F.; CORTEZ, DANIELA, V; GONCALVES, DANIEL B.; BENTO, HEITOR B. S.; GONCALVES, RHYAN L. N.; COSTA-SILVA, TALES A.; GAMBARATO, BRUNO C.; DE CASTRO, HEIZIR F.; DE CARVALHO, ANA KARINE F.. Biotechnological valorization of mycelium-bound lipase of Penicillium purpurogenum in hydrolysis of high content lauric acid vegetable oils. PROCESS SAFETY AND ENVIRONMENTAL PROTECTION, v. 161, p. 8-pg., 2022-05-01. (20/15513-7)

BENTO, HEITOR B. S.; REIS, CRISTIANO E. R.; CUNHA, PIETRO G.; CARVALHO, ANA K. F.; DE CASTRO, HEIZIR F.. One-pot fungal biomass-to-biodiesel process: Influence of the molar ratio and the concentration of acid heterogenous catalyst on reaction yield and costs. FUEL, v. 300, SEP 15 2021. (20/15513-7, 16/10636-8)

ZORN, SAVIENNE; CARVALHO, ANA; BENTO, HEITOR; GAMBARATO, BRUNO; PEDRO, GUILHERME; DA SILVA, ANA; GONCALVES, RHYAN; DA ROS, PATRICIA; SILVA, MESSIAS. Use of Fungal Mycelium as Biosupport in the Formation of Lichen-like Structure: Recovery of Algal Grown in Sugarcane Molasses for Lipid Accumulation and Balanced Fatty Acid Profile. MEMBRANES, v. 12, n. 3, p. 15-pg., 2022-03-01. (16/10636-8, 20/15513-7, 18/01386-3)

BARBIERI, GIORGIA S.; BENTO, HEITOR B. S.; DE OLIVEIRA, FERNANDA; PICHELI, FLAVIO P.; DIAS, LIDIA M.; MASARIN, FERNANDO; SANTOS-EBINUMA, VALERIA C.. Xylanase Production by Talaromyces amestolkiae Valuing Agroindustrial Byproducts. BIOTECH, v. 11, n. 2, p. 12-pg., 2022-05-17. (14/01580-3, 19/15493-9, 18/06908-8, 21/06686-8, 20/15513-7)

ZORN, SAVIENNE M. F. E.; DA SILVA, ANA PAULA T.; BREDDA, EDUARDO H.; BENTO, HEITOR B. S.; PEDRO, GUILHERME A.; CARVALHO, ANA KARINE F.; SILVA, MESSIAS BORGES; DA ROS, PATRICIA C. M.. In Situ Transesterification of Microbial Biomass for Biolubricant Production Catalyzed by Heteropolyacid Supported on Niobium. ENERGIES, v. 15, n. 4, p. 12-pg., 2022-02-01. (16/10636-8, 16/18909-3, 18/01386-3, 20/15513-7)

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)

DE LIMA, ROSEMAR; BENTO, HEITOR B. S.; REIS, CRISTIANO E. R.; VILAS BOAS, RENATA N.; DE FREITAS, LARISSA; CARVALHO, ANA K. F.; DE CASTRO, HEIZIR F.. Biolubricant Production from Stearic Acid and Residual Secondary Alcohols: System and Reaction Design for Lipase-Catalyzed Batch and Continuous Processes. CATALYSIS LETTERS, v. 152, n. 2, p. 547-558, FEB 2022. (16/10636-8, 20/15513-7)

CASTRO, DANIEL; MARQUES, ANA SOFIA C.; ALMEIDA, MAFALDA R.; DE PAIVA, GABRIELA B.; BENTO, HEITOR B. S.; PEDROLLI, DANIELLE B.; FREIRE, MARA G.; TAVARES, ANA P. M.; SANTOS-EBINUMA, VALERIA C.. L-asparaginase production review: bioprocess design and biochemical characteristics. Applied Microbiology and Biotechnology, v. 105, n. 11, p. 4515-4534, JUN 2021. (18/06908-8, 20/15513-7)

BENTO, HEITOR B. S.; REIS, CRISTIANO E. R.; PINTO, PEDRO A.; CORTEZ, DANIELA V.; VILAS BOAS, RENATA N.; COSTA-SILVA, TALES A.; CARVALHO, ANA K. F.; DE CASTRO, HEIZIR F.. Continuous Synthesis of Biodiesel from Outstanding Kernel Oil in a Packed Bed Reactor Using Burkholderia cepacia Lipase Immobilized on Magnetic Nanosupport. CATALYSIS LETTERS, OCT 2021. (20/15513-7)

MACHADO, SARA A.; RODRIGUES REIS, CRISTIANO E.; BENTO, HEITOR B. S.; CARVALHO, ANA K. F.; COSTA-SILVA, TALES A.; DA CONCEICAO, LEYVISON RAFAEL, V; GIORDANI, DOMINGOS S.; DE CASTRO, HEIZIR F.. Esterification of enzymatically treated macaw palm oil catalyzed by heteropoliacid supported onto Nb2O5. BIOFUELS-UK, v. 13, n. 8, p. 9-pg., 2022-04-27. (16/10636-8, 20/15513-7)