Genome editing of Bacillus subtilis for production of glutamine

• Grant number: 22/13103-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2022
• End date: February 29, 2024
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Danielle Biscaro Pedrolli
• Grantee: Leonardo Ferro Tavares
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The textile industry is one of the most profitable industries in the world. In its variety lines of manufactured types are pieces of fabrics and many types of dyes in the confections of types. One of the most famous and used blue dyes is indigo, produced from a reduction reaction through a process known as hydrosulfite vat solutions. The fast and efficient process is harmful to the environment, in particular due to the generation of toxic by-products. Not being the only major environmental problem caused by its use. Indigo itself is a harmful environment, especially to protected environments. Much of it is lost in the process and ends up in effluents going to water bodies and sources of all kinds. In them, it causes color changes that are responsible for reducing the incidence of sunlight in the water, compromising photosynthesis, increasing the COD of these ecosystems, fed by the heat available also for organized organisms. An alternative to indigo is the also natural blue dye, indigodine, naturally synthesized by some microorganisms. This presents problems associated with toxic by-products being a more sustainable and ecologically correct alternative. The obstacle to its large-scale production is the low natural productivity. As an alternative, there is heterologous production. The biobiointant is modified by the conversion of the enzyme by the modification of the enzyme that is altered through other processes of cells-glutamine in the cells, from a self-regulation by the change of cells-glutamine in the cells, from a self-regulation of its synthesis to the other cell-glutamine regulatory factors in cells, from a self-regulation of their synthesis to the regulation of the enzyme. The synthesizer is built and simplified by manufacturing the product without the need for mutation to target glutamine, the glnA protection for this project by the product and action on cell regulatory factors. The system in question will be built and cloned in Bacillus subtilis using the CRISPR-Cas9 gene editing technique by replacing the rocG gene. The execution of this is important in terms of several technical aspects associated with it, but also for the advancement of other projects related to it.