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Engineering of an endopolygalacturonase by insertion of different Carbohydrate Binding Modules (CBMs): chimerogenesis and proximity effects

Grant number: 17/13734-3
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): February 01, 2018
Effective date (End): October 31, 2020
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Richard John Ward
Grantee:Sibeli de Carli
Home Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated scholarship(s):19/26891-5 - Effects of the pectinolytic activity of endopolygacturonase-CBMs chimeras on the saccharification of different biomass feedstocks, BE.EP.DR


The large surplus of biomass resulting from processing sugarcane for 1st generation ethanol production, can be used for cogeneration of electricity or for the production of 2nd generation ethanol. Strategies to reduce the costs related to the enzymatic hydrolysis of these lignocellulosic residues include the identification and heterologous expression of enzymes with interesting catalytic characteristics, the use of protein engineering to improve their catalytic performance, and finally, the formulation of more efficient enzymatic cocktails. The controlled deconstruction of the plant cell wall lignocellulosic matrix by enzymes is challenging and constitutes a major barrier to achieve high-yield production of bioethanol at competitive prices. It is known that the presence of pectins, one of the main components of the middle lamella in plants, hinders the hydrolysis of the plant cell wall and prevents the efficient release of sugars. With the goal of developing technologies to optimize the hydrolysis of this polysaccharide, we intend to evaluate the effects of the proximity of different polysaccharides present in the plant cell wall on the activity of an endopolygalacturonase (endoPG I), engineered by the addition of a series of Carbohydrate Binding Modules (CBMs). Protein chimeras will be constructed from the fusion of endoPG I and different CBMs with specific affinities to cellulose, xylan, pectin, ²-glucan and xyloglucan. Following a biochemical characterization using a range of analytic techniques, these chimeras will be used to supplement enzymatic cocktails for the hydrolysis of lignocellulosic residues, such as straw and sugarcane bagasse, with the goal of increasing the yields of fermentable sugars. (AU)

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Scientific publications (4)
(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)
CARLI, SIBELI; MELEIRO, LUANA PARRAS; WARD, RICHARD JOHN. Biochemical and kinetic characterization of the recombinant GH28 Stereum purpureum endopolygalacturonase and its biotechnological application. International Journal of Biological Macromolecules, v. 137, p. 469-474, SEP 15 2019. Web of Science Citations: 0.
CARLI, SIBELI; BUFFONI DE CAMPOS CARNEIRO, LARA APARECIDA; WARD, RICHARD JOHN; MELEIRO, LUANA PARRAS. Immobilization of a beta-glucosidase and an endoglucanase in ferromagnetic nanoparticles: A study of synergistic effects. Protein Expression and Purification, v. 160, p. 28-35, AUG 2019. Web of Science Citations: 2.
SANTOS SALGADO, JOSE CARLOS; MELEIRO, LUANA PARRAS; CARLI, SIBELI; WARD, RICHARD JOHN. Glucose tolerant and glucose stimulated beta-glucosidases - A review. Bioresource Technology, v. 267, p. 704-713, NOV 2018. Web of Science Citations: 15.
DE CARVALHO, DANIELLA ROMANO; CARLI, SIBELI; MELEIRO, LUANA PARRAS; ROSA, JOSE CESAR; CAVALCANTE DE OLIVEIRA, ARTHUR HENRIQUE; JORGE, JOAO ATILIO; MELO FURRIEL, ROSA PRAZERES. A halotolerant bifunctional beta-xylosidase/alpha-L-arabinofuranosidase from Colletotrichum graminicola: Purification and biochemical characterization. International Journal of Biological Macromolecules, v. 114, p. 741-750, JUL 15 2018. Web of Science Citations: 7.

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