Recombinant expression of a Citrus miraculin in Pichia pastoris

• Grant number: 21/00718-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2021
• End date: December 31, 2022
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Andrea Soares da Costa Fuentes
• Grantee: Isabela Giacomeli de Deus
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

The high consumption of sugar in Brazil and in the world has had drastic consequences for human health, bringing diseases such as obesity and related diseases such as diabetes and metabolic syndrome. In the search for alternatives to decrease in sugar consumption, several studies have emerged related to the development of caloric and non-caloric sweeteners, of natural or artificial origin. Among the natural compounds there is a group of proteins that have the property of modifying the taste of food. These proteins are called Taste-Modifying Proteins (TMP). Within this class of molecules, there is the protein called miraculin, typical of the Richardella dulcifica plant, native to Africa, which has the property of modifying the acidic flavor of sweet foods. Due to this potential, several trials have been reported carried out in an attempt to produce miraculin through recombinant expression in other organisms, not obtaining considerable success for commercialization. However, the discovery of miraculin-like proteins in other plants has enabled further studies, although there is still no description of miraculin-like proteins with taste-modifying properties. In contrast, in silico analyzes allowed to find in Citrus paradisi, a protein with a gene sequence with high similarity to that found in the original miraculin. Therefore, the characterization of this protein and the exploration of its potential taste modification capacity can result in non-caloric alternatives to sweeten acidic foods, thus contributing to an improvement in human health. Therefore, the aim was the recombinant expression of a miraculin from Citrus paradisi, called Citros_Mir, in Pichia pastoris.