Design and Characterization of Multifunctional Xylanolytic Enzymes

• Grant number: 16/24139-6
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2017
• End date: September 30, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Richard John Ward
• Grantee: Richard John Ward
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 19/05276-0 - Technical laboratory support for the production and purification of proteins, BP.TT; 18/02786-5 - Design and characterization of multifunctional xylanolytic enzymes, BP.TT

Abstract

Biodegradation of lignocellulose has been the focus of intense study because of its applicability in sustainable biotechnological processes. The use of enzymes for the processing of lignocellulosic material is a key step not only in the production of second generation bioethanol, but also presents an opportunity to reduce input and energy costs in the pulp and paper industry, in the food industry and in biorefinery. Enzymes represents a significant fraction of the costs associated with the saccharification of lignocellulosic material, and protein engineering strategies can be applied to improve the performance of these biocatalysts. The current project proposes to use rational and semi-rational methods of protein design to create multifunctional chimeric enzymes to act synergistically in the degradation of arabinoxylan, one of the most abundant polysaccharides in the cell wall of plants and a source of C5 and C6 sugars. The project involves a two-stage experimental strategy; 1) evaluate the products generated from the action of mixtures of xylyanalytic enzymes against the arabinoxylan substrate using LC-MS and HPAEC-PAD. This step will identify complementary enzymatic activities, as well as the combinations of enzymes that demonstrate synergistic activities, and 2) application of rational and semi-rational protein design methodologies to create multifunctional enzymes using a beta-prism scaffold protein designed ab initio in the LBBP, together with the characterization of the catalytic properties of the chimeric enzymes. (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)

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)

BARRETO, MATHEUS QUINTANA; GARBELOTTI, CAROLINA VICTAL; SOARES, JESSICA DE MOURA; GRANDIS, ADRIANA; BUCKERIDGE, MARCOS SILVEIRA; LEONE, FRANCISCO ASSIS; WARD, RICHARD JOHN. Xylose isomerase from Piromyces sp. E2 is a promiscuous enzyme with epimerase activity. Enzyme and Microbial Technology, v. 166, p. 9-pg., 2023-03-24. (16/24139-6, 14/50884-5, 18/25664-2)

PINHEIRO, P. MATHEUS; REIS, A. G. RENATA; DUPREE, PAUL; WARD, J. RICHARD. Plant cell wall architecture guided design of CBM3-GH11 chimeras with enhanced xylanase activity using a tandem repeat left-handed beta-3-prism scaffold. COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL, v. 19, p. 1108-1118, 2021. (12/01066-2, 16/24139-6, 14/13956-8, 14/50884-5)

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)

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)

GARBELOTTI, CAROLINA VICTAL; GRANDIS, ADRIANA; CREVELIN, EDUARDO; BUCKERIDGE, MARCOS SILVEIRA; DE MORAES, LUIZ ALBERTO BERALDO; WARD, RICHARD JOHN. Glycomic profiling identifies key-structural differences in three arabinoxylan fractions from sugarcane culms. Carbohydrate Polymers, v. 310, p. 13-pg., 2023-02-24. (14/50884-5, 16/24139-6, 19/13936-0)

SOARES, JESSICA M.; CARNEIRO, LARA A. B. C.; BARRETO, MATHEUS Q.; WARD, RICHARD J.. Co-immobilization of multiple enzymes on ferromagnetic nanoparticles for the depolymerization of xyloglucan. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, v. 16, n. 6, p. 14-pg., 2022-08-02. (16/24139-6, 17/14452-1, 14/50884-5, 12/24147-8, 16/15708-7)

BUFFONI DE CAMPOS CARNEIRO, LARA APARECIDA; WURMAN, JOEL; DUPREE, PAUL; WARD, RICHARD JOHN. The Glycoside Hydrolase Family 35 beta-galactosidase from Trichoderma reesei debranches xyloglucan oligosaccharides from tamarind and jatoba. Biochimie, v. 211, p. 9-pg., 2023-02-27. (14/50884-5, 16/24139-6, 12/24147-8, 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)