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

• Grant number: 21/09659-1
• Support type: Scholarships abroad - Research Internship - Doctorate (Direct)
• Effective date (Start): November 01, 2021
• Effective date (End): September 01, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal researcher: Leticia Maria Zanphorlin
• Grantee: Renan Yuji Miyamoto
• Supervisor abroad: Volker Sieber
• Home Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovações (Brasil). Campinas , SP, Brazil
• Research place: 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 othervitamin. However, although vitamin B6 is essential, humans cannot synthesize it, so thismicronutrient 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 biologicalroutes still suffer to produce high titers due to toxic features of its intermediates, and itsfinal product can interact with a wide range of metabolic reactions. Thus, we propose abio-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 thiscan be achieved through mutual knowledge exchange, where the Brazilian enzymaticbackground (Mr. Miyamoto and Dr. Zanphorlin) will be deeply enhanced by the Germanknow-how on the cell-free bioprocess design (Dr. Sieber group). For this project, it wasset 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 thebioprocess through Placket-Burmann analysis and rotatable central composite design.Therefore, we expect to apply novel synthetic biochemistry techniques to develop a novelapproach for vitamin B6 production to overcome the major drawbacks faced by state-ofarttechnologies using xylose as raw-material.