Enzymatic cascade synthesis of vitamin B6 through a xylose-based route

• Grant number: 21/09659-1
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: November 01, 2021
• End date: September 01, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Leticia Maria Zanphorlin
• Grantee: Renan Yuji Miyamoto
• Supervisor: Volker Sieber
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Campinas , SP, Brazil
• Institution abroad: Technical University of Munich, Straubing (TUM), Germany
• Associated to the scholarship: 18/02865-2 - Molecular investigation of new xylose isomerases for application to lignocellulosic materials fermentation, BP.DD

Abstract

Vitamin B6 is a fundamental nutrient involved in more bodily processes than any other vitamin. However, although vitamin B6 is essential, humans cannot synthesize it, so this micronutrient must be obtained from foods or isolated supplements. Currently, vitaminB6 may be produced by technological routes through chemical or fermentative processes. Chemical synthesis employs expensive and/or hazardous resources while biological routes still suffer to produce high titers due to toxic features of its intermediates, and its final product can interact with a wide range of metabolic reactions. Thus, we propose a bio-based production strategy via a synthetic biochemistry approach to make vitamin B6in vitro from a xylose-conversion enzyme cascade through six steps. We hope that this can be achieved through mutual knowledge exchange, where the Brazilian enzymatic background (Mr. Miyamoto and Dr. Zanphorlin) will be deeply enhanced by the German know-how on the cell-free bioprocess design (Dr. Sieber group). For this project, it was set three main objectives (I) synthesize all six genes and express in the soluble fraction;(II) Test all purified enzymes in one-pot reaction; and (III) evaluate and optimize the bioprocess through Placket-Burmann analysis and rotatable central composite design. Therefore, we expect to apply novel synthetic biochemistry techniques to develop a novel approach for vitamin B6 production to overcome the major drawbacks faced by state-of art technologies using xylose as raw-material. (AU)