Molecular investigation of new xylose isomerases for application to lignocellulosic materials fermentation

• Grant number: 18/02865-2
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: August 01, 2018
• End date: August 31, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Leticia Maria Zanphorlin
• Grantee: Renan Yuji Miyamoto
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 15/26982-0 - Exploring novel strategies for depolymerization of plant cell-wall polysaccharides: from structure, function and rational design of glycosyl hydrolases to biological implications and potential biotechnological applications, AP.TEM
• Associated scholarship(s): 21/09659-1 - Enzymatic cascade synthesis of vitamin B6 through a xylose-based route, BE.EP.DD

Abstract

To become the second-generation ethanol production economically viable, it is crucial the hemicellulose fraction utilization, composed by pentoses, mainly xylose. Xylose, obtained by saccharification of hemicellulosic structures, is currently underappreciated, chiefly because the industrial yeasts, Saccharomyces cerevisiae, can not metabolize this sugar to produce second generation ethanol (E2G). Thus, a strategy to improve xylose utilization by S. cerevisiae consists in isomerization of xylose to xylulose by enzymes named Xylose Isomerase (XylA). However, most of XylA presents optimum pH and temperatures incompatible with physiological condition of S. cerevisiae, besides present very inefficient expression levels. In this scenario, this project aims the investigation of new xylose isomerase that was previously selected by in silico approach, to operate in fermentation processes with industrial yeast Saccharomyces cerevisiae. The XylA will be expressed in E. coli BL21 and will be evaluated using a multidisciplinary approach that includes biochemical and classical biophysical assays, as well as cristalization, and solve the tertiary structure, besides ex-situ Simultaneous Isomerization and Fermentation (SIF). In the end, the XylA that presents more efficient and compatible with fermentation conditions will be had it codon optimized and transformed in S. cerevisiae. The Genetically Modified Microrganism (GMO) Saccharomyces cerevisiae developed will be evaluated against fermentation assays with xylose and then compared with the one already used industrially. (AU)

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)

DE SOUSA, AMANDA S.; DE MELO, RICARDO R.; MIYAMOTO, RENAN Y.; MORAIS, MARIANA A. B.; ANDRADE, LILIANE P.; MILAN, NATALIA; DE AVILA, MAYARA C.; DE SOUZA, CLAUDIA M.; ADAO, REGINA C.; SCARPASSA, JOSIANE A.; et al. A rationally identified marine GH1 beta-glucosidase has distinguishing functional features for simultaneous saccharification and fermentation. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, AUG 2020. (16/19995-0, 17/14253-9, 18/02865-2, 15/26982-0, 16/06509-0, 19/08855-1)

MIYAMOTO, RENAN YUJI; DE MELO, RICARDO RODRIGUES; DE MESQUITA SAMPAIO, ISABELLE LOBO; DE SOUSA, AMANDA SILVA; MORAIS, EDVALDO RODRIGO; SARGO, CINTIA REGINA; ZANPHORLIN, LETICIA MARIA. Paradigm shift in xylose isomerase usage: a novel scenario with distinct applications. CRITICAL REVIEWS IN BIOTECHNOLOGY, OCT 2021. (19/08855-1, 17/14253-9, 18/02865-2)

XAVIER GUILHERME, EDERSON PAULO; ZANPHORLIN, LETICIA MARIA; SOUSA, AMANDA SILVA; MIYAMOTO, RENAN YUJI; OLIVEIRA BRUZIQUESI, CARLOS GIOVANI; APARECIDA DE CARVALHO MESQUITA, BRUNA MARA; SOUSA SANTOS, SERGIO HENRIQUE; AGUIAR-OLIVEIRA, ELIZAMA; COTA, JUNIO. Simultaneous saccharification isomerization and Co-fermentation - SSICF: A new process concept for second-generation ethanol biorefineries combining immobilized recombinant enzymes and non-GMO Saccharomyces. RENEWABLE ENERGY, v. 182, p. 274-284, JAN 2022. (18/02865-2, 19/08855-1)

DE SOUSA, AMANDA S.; DE MELO, RICARDO R.; MIYAMOTO, RENAN Y.; MORAIS, MARIANA A. B.; ANDRADE, LILIANE P.; MILAN, NATALIA; DE AVILA, MAYARA C.; DE SOUZA, CLAUDIA M.; ADAO, REGINA C.; SCARPASSA, JOSIANE A.; et al. A rationally identified marine GH1 beta-glucosidase has distinguishing functional features for simultaneous saccharification and fermentation. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, v. 14, n. 6, p. 17-pg., 2020-08-20. (19/08855-1, 17/14253-9, 18/02865-2, 15/26982-0, 16/06509-0, 16/19995-0)