Development of strategies for immobilization and co-immobilization of enzymes involved in biomass degradation

• Grant number: 20/00081-4
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): March 01, 2020
• Effective date (End): March 31, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Acordo de Cooperação: Fundação para a Ciência e a Tecnologia (FCT)
• Principal Investigator: Maria de Lourdes Teixeira de Moraes Polizeli
• Grantee: Robson Carlos Alnoch
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 18/07522-6 - Development of eco-friendly technologies for the hydrolysis of biomass and residues of cellulose pulp and paper industry, AP.R

Abstract

During the hydrolysis process of the biomass one of the critical points is the rapid degradation of the enzymes. Therefore, techniques to stabilize and reduce inactivation of enzymes, due to the distortion of their native structure by the influence of temperature, pH and organic solvents, are quite attractive. Immobilization of enzymes has been employed for this purpose, making the process less costly. This strategy is also an important tool to increase its bioactivity, allowing it to be reused. Immobilization is a method of fixing the enzyme to an immobilizing agent, and this method should be as smooth as possible, avoiding that the enzyme undergoes conformational change. The support of immobilization has an important role in maintaining the actual conformation and activity of the enzyme. Despite the great diversity of methods developed and applied in the immobilization of enzymes, there is no effective method for all of them, and the work of immobilization of cellulases and xylanases for the hydrolysis of lignocellulosic biomass is scarce. Among the different methods, immobilization by covalent bonding and ion exchange has been widely employed by increasing the resistance of enzymes to high temperatures and denaturing solvents. The best-known immobilization techniques are: glyoxyl-agarose, MANAE-agarose, glutaraldehyde-chitosan, betwixt others. Therefore, in this project will be used substrates based on agarose, porous supports, carbon nanotubes and supports produced from the biomass itself. In this context, it will evaluate covalent and ion exchange immobilization techniques that result in an enzyme-immobilizing agent complex with good activity retention and high operational stability. Co-immobilization processes will also be evaluated to immobilize two or more enzymes. The evaluation of the different treatments and the capacity of the immobilization techniques will be performed by determining the immobilization efficiency, stability and bioactivity of the immobilized enzymes and co-immobilized in different chemical supports, as well as to evaluate the biocatalytic reusability. This project opens the perspective to develop new methodologies for the immobilization and co-immobilization of enzymes that aim at the degradation of different biomasses. In this work, it will have demonstrated methods for immobilization, using different material for support. According to each enzymes characteristic it will be followed different strategies for immobilization and co-immobilization. (AU)

Scientific publications

(9)

(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)

COELHO MOREIRA, MICHELLE APARECIDA; ALNOCH, ROBSON CARLOS; DE LIMA LUZ JUNIOR, LUIZ FERNANDO; BARON, ALESSANDRA MACHADO; MITCHELL, DAVID ALEXANDER; KRIEGER, NADIA. Transesterification of castor oil catalyzed by a fermented solid produced by Burkholderia contaminans using a bioreactor coupled with ultrasound irradiation. Biotechnology and Applied Biochemistry, v. N/A, p. 11-pg., 2022-10-24. (20/00081-4)

MALDONADO, MARCOS RODRIGUES; ALNOCH, ROBSON CARLOS; DE ALMEIDA, JANAINA MARQUES; DOS SANTOS, LEANDRO ALVES; ANDRETTA, ANDRESSA TEDESCO; PILAR CUASPA ROPAIN, ROCIO DEL; DE SOUZA, EMANUEL MALTEMPI; MITCHELL, DAVID ALEXANDER; KRIEGER, NADIA. ey mutation sites for improvement of the enantioselectivity of lipases through protein engineerin. Biochemical Engineering Journal, v. 172, AUG 2021. (20/00081-4)

DESAGIACOMO, CARLA CRISTINA VILLELA; ALNOCH, ROBSON CARLOS; PINHEIRO, VANESSA ELISA; CEREIA, MARIANA; MACHADO, CARLA BOTELHO; DAMASIO, ANDRE; AUGUSTO, MARLEI JOSIELE; PEDERSOLI, WELLINGTON; SILVA, ROBERTO NASCIMENTO; POLIZELI, MARIA DE LOURDES TEIXEIRA DE MORAES. Structural model and functional properties of an exo-polygalacturonase from Neosartorya glabra. International Journal of Biological Macromolecules, v. 186, p. 909-918, SEP 1 2021. (14/50884-5, 20/00081-4, 18/07522-6, 17/22669-0)

DE FREITAS, EMANUELLE NEIVERTH; ALNOCH, ROBSON CARLOS; CONTATO, ALEX GRACA; NOGUEIRA, V, KAROLINE MARIA; CREVELIN, EDUARDO JOSE; BERALDO DE MORAES, LUIZ ALBERTO; SILVA, ROBERTO NASCIMENTO; MARTINEZ, CARLOS ALBERTO; POLIZELI, MARIA DE LOURDES T. M.. Enzymatic Pretreatment with Laccases from Lentinus sajor-caju Induces Structural Modification in Lignin and Enhances the Digestibility of Tropical Forage Grass (Panicum maximum) Grown under Future Climate Conditions. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 22, n. 17, SEP 2021. (18/07522-6, 19/07141-5, 08/57908-6, 17/23989-9, 08/58075-8, 20/00081-4)

ALNOCH, ROBSON CARLOS; DOS SANTOS, LEANDRO ALVES; DE ALMEIDA, JANAINA MARQUES; KRIEGER, NADIA; MATEO, CESAR. Recent Trends in Biomaterials for Immobilization of Lipases for Application in Non-Conventional Media. CATALYSTS, v. 10, n. 6, JUN 2020. (20/00081-4, 18/07522-6)

ALNOCH, ROBSON C.; ALVES, GABRIELA S.; SALGADO, JOSE C. S.; DE ANDRADES, DIANDRA; DE FREITAS, EMANUELLE N.; NOGUEIRA, KAROLINE M. V.; VICI, ANA C.; OLIVEIRA, DOUGLAS P.; CARVALHO-JR, VALDEMIRO P.; SILVA, ROBERTO N.; et al. Immobilization and Application of the Recombinant Xylanase GH10 of Malbranchea pulchella in the Production of Xylooligosaccharides from Hydrothermal Liquor of the Eucalyptus (Eucalyptus grandis) Wood Chips. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 23, n. 21, p. 18-pg., 2022-11-01. (14/50884-5, 18/07522-6, 19/21989-7, 20/15510-8, 20/00081-4)

MALDONADO, MARCOS RODRIGUES; ALNOCH, ROBSON CARLOS; SHIRATORI, GABRIELA YUMI YAMAMOTO; DE OLIVEIRA, CYNTHIA CHRISTINA; GONCALVES, MARCOS BROWN; MITCHELL, DAVID ALEXANDER; KRIEGER, NADIA. Modification of the properties of the metagenomic lipase LipC12 by engineering of the hydrophobic cavity. BIOCATALYSIS AND BIOTRANSFORMATION, v. N/A, p. 11-pg., 2023-05-02. (20/00081-4)

MALDONADO, MARCOS RODRIGUES; ALNOCH, ROBSON CARLOS; DE ALMEIDA, JANAINA MARQUES; DOS SANTOS, LEANDRO ALVES; ANDRETTA, ANDRESSA TEDESCO; PILAR CUASPA ROPAIN, ROCIO DEL; DE SOUZA, EMANUEL MALTEMPI; MITCHELL, DAVID ALEXANDER; KRIEGER, NADIA. Key mutation sites for improvement of the enantioselectivity of lipases through protein engineering. Biochemical Engineering Journal, v. 172, p. 13-pg., 2021-05-06. (20/00081-4)

DOS SANTOS, LEANDRO ALVES; ALNOCH, ROBSON CARLOS; SOARES, GLEYDISON AMARANTE; MITCHELL, DAVID ALEXANDER; KRIEGER, NADIA. Immobilization of Pseudomonas fluorescens lipase on chitosan crosslinked with polyaldehyde starch for kinetic resolution of sec-alcohols. Process Biochemistry, v. 122, p. 10-pg., 2022-10-20. (20/00081-4)