Development of sustainable nanomaterials for the purification of antileukemic drugs

• Grant number: 18/06908-8
• Support Opportunities: Regular Research Grants
• Start date: September 01, 2018
• End date: August 31, 2022
• Field of knowledge: Engineering - Chemical Engineering - Chemical Process Industries
• Agreement: Fundação para a Ciência e a Tecnologia (FCT)
• Principal Investigator: Valéria de Carvalho Santos Ebinuma
• Grantee: Valéria de Carvalho Santos Ebinuma
• Principal researcher abroad: Ana Paula Mora Tavares
• Institution abroad: Universidade de Aveiro (UA), Portugal
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated researchers: Adalberto Pessoa Junior ; Cláudia Sofia Castro Gomes da Silva ; Danielle Biscaro Pedrolli ; Jorge Pereira
• Associated scholarship(s): 20/15513-7 - Scale up of L-asparaginase production employing genetically modified microorganism, BP.PD; 19/01757-4 - Production of L-asparaginase using genetically modified microorganism, BP.PD

Abstract

Getting older is the biggest risk factor for most fatal diseases, including cancer, heart disease and Alzheimer's. The advances in nanomedicine comprise the development of nanoparticles either for drug delivering or as platforms for biomolecules purification. Simultaneously, continuous progresses have been made for the development of recombinant therapeutic enzymes, namely L-asparaginase (LA), which presents antineoplastic properties for thetreatment of leukaemia's, in particular for Acute Lymphoblastic Leukemia (ALL). Currently, the LA available in medicalapplications is produced from bacteria. However, this type of LA may activate the immunological system causinghuman hypersensitivity reactions. Therefore, the search for novel microorganisms able to produce LA with no sideeffects has been the main focus of many researches.Considering the great demand for the development of new technologies for the purification of therapeutic enzymes,still limited by their high cost, this project aims to produce recombinant LA by bacteria fermentation and to developa cost-effective and sustainable platform for the biopharmaceutical purification (downstream processing) from the fermentation broth using nanomaterials. The main goal is to obtain recoveries of LA above 90% and purity levels over95%, with a cost reduction around 20%. A critical evaluation of the best strategies and conditions for the integrationprocesses and scale-up will be explored. The involved research teams (CICECO, FEUP and UNESP) have shownto be successful in providing pioneering scientific results on the production of enzymes and on the developmentof separation and purification techniques for biopharmaceuticals. The UNESP team from Brazil will optimise noveltechniques of heterologous enzyme production by bacterial fermentation, which is generally regarded as safe (GRAS)and easy to manipulate, in order to achieve higher enzyme yields. The Portuguese teams (CICECO and FEUP) willbe responsible for the preparation of functionalisation of silica and carbon nanomaterials, which will be used as anefficient and fast strategy for LA purification from the fermentation medium. NanoPurAsp aims to meet the approval of biopharmaceuticals by regulatory authorities which require effective andhigh quality biopharmaceuticals, i.e., no side-effects, high purity and biological activity. These can only be attained through in-depth knowledge of the products and of their manufacturing processes. The advanced research here produced will certainly allow the success of therapeutic enzymes production and its easy scalability up to industrial levels at a lower cost. The integrated process developed will have a great impact in economy with a long-term impact in human healthcare. This project aspires the optimization of resources and materials which will contribute to a greener and sustainable growth, which in turn will be a source of economic valorisation for the future industrial manufacturing. (AU)

Scientific publications

(20)

(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)

MUSSAGY, CASSAMO U.; PEREIRA, JORGE F. B.; DUFOSSE, LAURENT; RAGHAVAN, VIJAYA; SANTOS-EBINUMA, VALERIA C.; PESSOA JR, ADALBERTO. Advances and trends in biotechnological production of natural astaxanthin by Phaffia rhodozyma yeast. CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION, AUG 2021. (17/50303-0, 15/50058-0, 14/19793-3, 14/16424-7, 18/06908-8, 20/08655-0)

TACIN, MARIANA VENDRASCO; COSTA-SILVA, TALES A.; DE PAULA, ARIELA VELOSO; PALOMO, JOSE M.; SANTOS-EBINUMA, VALERIA DE CARVALHO. Microbial lipase: a new approach for a heterogeneous biocatalyst. PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, v. 51, n. 8, p. 749-760, SEP 1 2021. (14/01580-3, 18/06908-8)

CRISTOVAO, RAQUEL O.; BARROS, RITA A. M.; PINHO, JOAO G.; TEIXEIRA, LILIA S.; NEVES, MARCIA C.; FREIRE, MARA G.; FARIA, JOAQUIM L.; SANTOS-EBINUMA, VALERIA C.; TAVARES, ANA P. M.; SILVA, CLAUDIA G.. Unveiling the Influence of Carbon Nanotube Diameter and Surface Modification on the Anchorage of L-Asparaginase. APPLIED SCIENCES-BASEL, v. 12, n. 17, p. 17-pg., 2022-09-01. (18/06908-8)

BARROS, RITA A. M.; CRISTOVAO, RAQUEL O.; CARABINEIRO, SONIA A. C.; NEVES, MARCIA C.; FREIRE, MARA G.; FARIA, JOAQUIM L.; SANTOS-EBINUMA, VALERIA C.; TAVARES, ANA P. M.; SILVA, CLAUDIA G.. Immobilization and Characterization of L-Asparaginase over Carbon Xerogels. BIOTECH, v. 11, n. 2, p. 20-pg., 2022-04-14. (18/06908-8)

MUSSAGY, CASSAMO U.; FARIAS, FABIANE O.; SANTOS-EBINUMA, VALERIA C.; PEREIRA, JORGE F. B.; PESSOA JR, ADALBERTO. Sustainable one-pot platform for the green recovery of carotenoids from Phaffia rhodozyma yeast and their use as natural additives in soap formulation. ENVIRONMENTAL TECHNOLOGY & INNOVATION, v. 29, p. 12-pg., 2023-01-23. (18/06908-8, 20/08655-0, 21/06686-8)

VERISSIMO, NATHALIA VIEIRA; MUSSAGY, CASSAMO USSEMANE; OSHIRO, ARIANE ALVES; NOBREGA MENDONCA, CARLOS MIGUEL; SANTOS-EBINUMA, VALERIA DE CARVALHO; PESSOA JUNIOR, ADALBERTO; DE SOUZA OLIVEIRA, RICARDO PINHEIRO; BRANDAO PEREIRA, JORGE FERNANDO. From green to blue economy: Marine biorefineries for a sustainable ocean-based economy. GREEN CHEMISTRY, v. 23, n. 23, NOV 2021. (18/06908-8, 20/13271-6, 20/14144-8, 19/15493-9, 18/25511-1, 20/08655-0)

MUSSAGY, CASSAMO U.; SANTOS-EBINUMA, VALERIA C.; HERCULANO, RONDINELLI D.; COUTINHO, JOAO A. P.; PEREIRA, JORGE F. B.; PESSOA JR, ADALBERTO. Ionic liquids or eutectic solvents? Identifying the best solvents for the extraction of astaxanthin and beta-carotene from Phaffia rhodozyma yeast and preparation of biodegradable films. GREEN CHEMISTRY, v. 24, n. 1, NOV 2021. (19/15493-9, 15/11759-3, 18/06908-8, 20/08655-0)

REMONATTO, DANIELA; FERRARI, BARBARA RIBEIRO; BASSAN, JULIANA CRISTINA; MUSSAGY, CASSAMO USSEMANE; SANTOS-EBINUMA, VALERIA DE CARVALHO; DE PAULA, ARIELA VELOSO. Utilization of Clay Materials as Support for Aspergillus japonicus Lipase: An Eco-Friendly Approach. CATALYSTS, v. 11, n. 10, OCT 2021. (19/15493-9, 20/08655-0, 18/06908-8, 20/09592-1, 17/11482-7)

DE OLIVEIRA, FERNANDA; LIMA, CAIO DE AZEVEDO; LOPES, ANDRE MORENI; VIANA MARQUES, DANIELA DE ARAUJO; DRUZIAN, JANICE IZABEL; PESSOA JUNIOR, ADALBERTO; SANTOS-EBINUMA, VALERIA CARVALHO. Microbial Colorants Production in Stirred-Tank Bioreactor and Their Incorporation in an Alternative Food Packaging Biomaterial. JOURNAL OF FUNGI, v. 6, n. 4, DEC 2020. (18/06908-8, 18/10799-0, 17/10789-1, 14/01580-3, 13/05039-2)

MUSSAGY, CASSAMO U.; GONZALEZ-MIQUEL, MARIA; SANTOS-EBINUMA, VALERIA C.; PEREIRA, JORGE F. B.. Microbial torularhodin - a comprehensive review. CRITICAL REVIEWS IN BIOTECHNOLOGY, v. N/A, p. 19-pg., 2022-03-09. (18/06908-8, 20/08655-0, 21/06686-8)

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)

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)

VERISSIMO, NATHALIA VIEIRA; VICENTE, FILIPA A.; DE OLIVEIRA, RODRIGO CARDOSO; LIKOZAR, BLAZ; OLIVEIRA, RICARDO PINHEIRO DE SOUZA; PEREIRA, JORGE FERNANDO BRANDAO. Ionic liquids as protein stabilizers for biological and biomedical applications: A review. BIOTECHNOLOGY ADVANCES, v. 61, p. 29-pg., 2022-11-11. (18/25511-1, 19/15493-9, 20/14144-8, 20/10676-5, 18/06908-8)

TACIN, MARIANA VENDRASCO; COSTA-SILVA, TALES A.; DE PAULA, ARIELA VELOSO; PALOMO, JOSE M.; SANTOS-EBINUMA, VALERIA DE CARVALHO. Microbial lipase: a new approach for a heterogeneous biocatalyst. PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, NOV 2020. (18/06908-8, 14/01580-3)

CRISTOVAO, RAQUEL O.; ALMEIDA, MAFALDA R.; BARROS, MARIA A.; NUNES, JOAO C. F.; BOAVENTURA, RUI A. R.; LOUREIRO, JOSE M.; FARIA, JOAQUIM L.; NEVES, MARCIA C.; FREIRE, MARA G.; EBINUMA-SANTOS, VALERIA C.; et al. Development and characterization of a novell-asparaginase/MWCNT nanobioconjugate. RSC ADVANCES, v. 10, n. 52, p. 9-pg., 2020-08-25. (18/06908-8)

MUSSAGY, CASSAMO U.; KURNIA, KIKI A.; DIAS, ANA C. R. V.; RAGHAVAN, VIJAYA; SANTOS-EBINUMA, VALERIA C.; PESSOA JR, ADALBERTO. n eco-friendly approach for the recovery of astaxanthin and 8-carotene from Phaffia rhodozyma biomass using bio-based solvent. Bioresource Technology, v. 345, FEB 2022. (20/08655-0, 19/15493-9, 18/06908-8, 21/06686-8)

NUNES, JOAO C. F.; ALMEIDA, MAFALDA R.; BENTO, RUI M. F.; PEREIRA, MATHEUS M.; SANTOS-EBINUMA, VALERIA C.; NEVES, MARCIA C.; FREIRE, MARA G.; TAVARES, ANA P. M.. nhanced Enzyme Reuse through the Bioconjugation of L-Asparaginase and Silica-Based Supported Ionic Liquid-like Phase Material. OLECULE, v. 27, n. 3, FEB 2022. (18/06908-8)

ALMEIDA, MAFALDA R.; CRISTOVAO, RAQUEL O.; BARROS, MARIA A.; NUNES, JOAO C. F.; BOAVENTURA, RUI A. R.; LOUREIRO, JOSE M.; FARIA, JOAQUIM L.; NEVES, MARCIA C.; FREIRE, MARA G.; SANTOS-EBINUMA, VALERIA C.; et al. Superior operational stability of immobilized l-asparaginase over surface-modified carbon nanotubes. SCIENTIFIC REPORTS, v. 11, n. 1, NOV 2 2021. (18/06908-8)

DE OLIVEIRA, FERNANDA; ROCHA, INES L. D.; GOUVEIA ALVES PINTO, DIANA CLAUDIA; VENTURA, SONIA P. M.; DOS SANTOS, ANDRE GONZAGA; CREVELIN, EDUARDO JOSE; SANTOS EBINUMA, VALERIA DE CARVALHO. Identification of azaphilone derivatives of Monascus colorants from Talaromyces amestolkiae and their halochromic properties. Food Chemistry, v. 372, MAR 15 2022. (14/01580-3, 18/06908-8, 19/15493-9)

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)