Development and in silico evaluation of an expression platform based in E. coli for the production of enzymes involved in lignocellulosic biomass degradation

• Grant number: 14/13974-6
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: November 01, 2014
• End date: August 31, 2018
• Field of knowledge: Engineering - Chemical Engineering - Chemical Process Industries
• Principal Investigator: Adriano Rodrigues Azzoni
• Grantee: Rafael da Gama Ferreira
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Conversion of lignocellulosic biomass into fermentable sugars by enzymatic hydrolysis still represents a high cost step for production of second generation ethanol. To overcome this drawback, it is important to increase the enzyme efficiency and decrease production costs by using genetic engineering tools and process optimization. Hence, the development of more efficient heterologous expression systems that complement conventional systems (usually filamentous fungi) represents a subject of interest for making cellulosic ethanol viable. The present project aims to develop and evaluate an expression platform of recombinant enzymes using E. coli. Through molecular biology techniques, new strains that dispense with using antibiotics in the medium will be developed. The cell culture free of antibiotics will employ an E. coli strain (SE1) which is capable of maintaining the plasmid segregational stability by means of an "antidote-toxin" system. Using modified strains capable of expressing model enzymes (endo-1,4-beta-glucanase, endo-1,4-beta-xylanase, beta-xylosidase), critical parameters of fermentation will be obtained, such as the specific growth rate and the product (enzyme) biosynthesis rate, under high cell density. These parameters will be compared with those of conventional strains (BL21(DE3)), enabling us to carry out a detailed techno-economic and environmental evaluation using simulation software SuperPro Designer". Therefore, we expect to achieve, besides developing new E. coli strains capable of producing enzymes without antibiotics, a rigorous evaluation of economic and environmental impact of this new technology in the production of recombinant proteins, thus generating relevant knowledge for making second generation ethanol viable in Brazil. (AU)

Scientific publications

(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)

FERREIRA, RAFAEL G.; AZZONI, ADRIANO R.; FREITAS, SINDELIA. On the production cost of lignocellulose-degrading enzymes. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, v. 15, n. 1, p. 85-99, JAN 2021. (14/13974-6)

FERREIRA, RAFAEL DA GAMA; AZZONI, ADRIANO RODRIGUES; FREITAS, SINDELIA. Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant beta-glucosidase. BIOTECHNOLOGY FOR BIOFUELS, v. 11, MAR 29 2018. (10/08089-2, 14/13974-6)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

FERREIRA, Rafael da Gama. Development and in silico evaluation of an expression platform based on E.coli for the production of a recombinant beta-glucosidase.. 2019. Doctoral Thesis - Universidade de São Paulo (USP). Escola Politécnica (EP/BC) São Paulo.