Thermostability modulation of a ß-glucosidase stimulated by glucose and xylose using site directed glycosylation

• Grant number: 16/17582-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: April 01, 2017
• End date: January 31, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Richard John Ward
• Grantee: Luana Parras Meleiro Garcia
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 19/17958-9 - Thermostability modulation of a ²-glucosidase stimulated by glucose and xylose using site directed glycosylation: study of glycosylation pattern by LC-MS/MS, BE.EP.PD

Abstract

The depletion of global fossil fuel reserves together with increasing environmental concerns arising from use of these fuels has stimulated considerable interest in the production of cellulosic ethanol. After decades of research, however, the knowledge required to develop a process for the total enzymatic saccharification of lignocellulosic residues is still incomplete. One of the principal challenges is to overcome the costs associated with the use of enzymes in the process, thereby improving its economic viability. This is due to the fact that most enzymes do not satisfy all the characteristics necessary for the process, such as optimum temperature and pH reaction, thermal and pH stability, high catalytic efficiency for hydrolysis of the natural substrate, the possibility of high level expression of active enzymes in appropriate hosts and tolerance to products and reaction byproducts. Protein engineering has been widely employed for enzyme improvement, and in this project, we intend to use a relatively new approach, known as site directed glycosylation to improve the characteristics of a well studied enzyme that presents many of the desirable catalytic characteristics outlined above. With the aim of improving the thermostability of the ß-glucosidase from Humicola insolens (Bglhi), we intend to engineer the enzyme by modulation of its carbohydrate content. The expected results will help to clarify the factors that affect the thermal stabilization of proteins, and contribute to the development of more efficient enzyme cocktails for hydrolysis of lignocellulosic biomass. (AU)

Scientific publications

(11)

(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; 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. (17/13734-3, 16/17582-0)

CARLI, SIBELI; SALGADO, JOSE CARLOS SANTOS; MELEIRO, LUANA PARRAS; WARD, RICHARD JOHN. ovalent Immobilization of Chondrostereum purpureum Endopolygalacturonase on Ferromagnetic Nanoparticles: Catalytic Properties and Biotechnological Applicatio. Applied Biochemistry and Biotechnology, v. 194, n. 2, SEP 2021. (19/17958-9, 14/50884-5, 17/13734-3, 19/21989-7, 16/17582-0)

FURTADO, GILVAN PESSOA; CARLI, SIBELI; MELEIRO, LUANA PARRAS; SANTOS SALGADO, JOSE CARLOS; WARD, RICHARD JOHN. Enhanced hydrolytic efficiency of an engineered CBM11-glucanase enzyme chimera against barley beta-D-glucan extracts. Food Chemistry, v. 365, DEC 15 2021. (16/17582-0, 16/24139-6, 17/13734-3, 14/50884-5, 19/21989-7)

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. (17/13734-3, 16/17582-0)

ALVES, LUANA DE FATIMA; MELEIRO, LUANA PARRAS; SILVA, ROBERTO N.; WESTMANN, CAUA ANTUNES; GUAZZARONI, MARIA-EUGENIA. Novel Ethanol- and 5-Hydroxymethyl Furfural-Stimulated beta-Glucosidase Retrieved From a Brazilian Secondary Atlantic Forest Soil Metagenome. FRONTIERS IN MICROBIOLOGY, v. 9, OCT 29 2018. (16/05472-6, 15/04309-1, 16/06323-4, 16/17582-0)

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. (17/13734-3, 14/50884-5, 16/17582-0)

MELEIRO, LUANA PARRAS; SANTOS SALGADO, JOSE CARLOS; MALDONADO, RAQUEL FONSECA; CARLI, SIBELI; BERALDO MORAES, LUIZ ALBERTO; WARD, RICHARD JOHN; JORGE, JOAO ATILIO; MELO FURRIEL, ROSA PRAZERES. Engineering the GH1 beta-glucosidase from Humicola insolens: Insights on the stimulation of activity by glucose and xylose. PLoS One, v. 12, n. 11, NOV 16 2017. (10/18850-2, 16/17582-0, 14/14415-0)

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. (17/13734-3, 14/14415-0, 16/17582-0)

BUFFONI DE CAMPOS CARNEIRO, LARA APARECIDA; FUZO, CARLOS ALESSANDRO; MELEIRO, LUANA PARRAS; CARLI, SIBELI; BARRETO, MATHEUS QUINTANA; LOURENZONI, MARCOS ROBERTO; BUCKERIDGE, MARCOS SILVEIRA; WARD, RICHARD JOHN. Selective xyloglucan oligosaccharide hydrolysis by a GH31 alpha-xylosidase from Escherichia coli. Carbohydrate Polymers, v. 284, p. 12-pg., 2022-05-15. (12/24147-8, 17/13734-3, 16/24139-6, 16/17582-0, 14/50884-5, 16/15708-7)

TRINCA, VITOR; CARLI, SIBELI; ULIANA, JOAO VITOR CARDOSO; GARBELOTTI, CAROLINA VICTAL; DA SILVA, MARIANA MENDES; KUNES, VITOR; MELEIRO, LUANA PARRAS; BRANCINI, GUILHERME THOMAZ PEREIRA; MENZEL, FRANK; ANDRIOLI, LUIZ PAULO MOURA; et al. Biocatalytic potential of Pseudolycoriella CAZymes (Sciaroidea, Diptera) in degrading plant and cell wall. ISCIENCE, v. 26, n. 4, p. 27-pg., 2023-04-21. (14/14318-5, 20/05636-4, 16/17582-0, 16/24139-6, 16/25325-8, 17/13734-3, 18/25664-2)

CARLI, SIBELI; PARRAS MELEIRO, LUANA; SALGADO, JOSE CARLOS SANTOS; WARD, RICHARD JOHN. Synthetic carbohydrate-binding module-endogalacturonase chimeras increase catalytic efficiency and saccharification of lignocellulose residues. BIOMASS CONVERSION AND BIOREFINERY, v. N/A, p. 12-pg., 2022-05-03. (16/24139-6, 16/17582-0, 17/13734-3, 19/21989-7, 14/50884-5)