TY - JOUR
T1 - Biotechnological production of reduced and oxidized NAD+ precursors
AU - Zapata-Pérez, Rubén
AU - García-Saura, Antonio Ginés
AU - Scantlebery, Angelique M. L.
AU - Schomakers, Bauke V.
AU - Rabadán-Ros, Rubén
AU - van Weeghel, Michel
AU - Houtkooper, Riekelt H.
AU - Sánchez-Ferrer, Álvaro
N1 - Funding Information: RZP and RRR group is supported by grants from the Plan Propio de Ayuda a la Investigación 2020–21 – Programa de Apoyo a los Grupos de Investigación, Universidad Católica San Antonio de Murcia (PMAFI 25/21 and PMAFI 26/21), and from Fundación Séneca - Agencia de Ciencia y Tecnología de la Región de Murcia (22011/JLI/22, Ayudas a Proyectos para la Generación de Nuevo Liderazgo Científico Jóvenes Líderes en Investigación 2022). Work in the Houtkooper group is financially supported by an ERC Starting grant (no. 638290), a VIDI grant from ZonMw (no. 91715305), and a grant from the Velux Stiftung (no. 1063). ASF group is supported by Spanish grants from MINECO-FEDER (BIO2013-45336-R) and from Ayudas a los Grupos y Unidades de Excelencia Científica de la Región de Murcia, Fundación Séneca – Agencia de Ciencia y Tecnología de la Región de Murcia (19893/GERM/15, Programa de Apoyo a la Investigación 2014). Publisher Copyright: © 2023 Elsevier Ltd
PY - 2023/3/1
Y1 - 2023/3/1
N2 - Dysregulation of nicotinamide adenine dinucleotide (NAD+) homeostasis by increased activity of NAD+ consumers or reduced NAD+ biosynthesis plays an important role in the onset of prevalent, often age-related, diseases, such as diabetes, neuropathies or nephropathies. To counteract such dysregulation, NAD+ replenishment strategies can be used. Among these, administration of vitamin B3 derivatives (NAD+ precursors) has garnered attention in recent years. However, the high market price of these compounds and their limited availability, pose important limitations to their use in nutritional or biomedical applications. To overcome these limitations, we have designed an enzymatic method for the synthesis and purification of (1) the oxidized NAD+ precursors nicotinamide mononucleotide (NMN) and nicotinamide riboside (NR), (2) their reduced forms NMNH and NRH, and (3) their deaminated forms nicotinic acid mononucleotide (NaMN) and nicotinic acid riboside (NaR). Starting from NAD+ or NADH as substrates, we use a combination of three highly overexpressed soluble recombinant enzymes; (a) a NAD+ pyrophosphatase, (b) an NMN deamidase, and (c) a 5′-nucleotidase, to produce these six precursors. Finally, we validate the activity of the enzymatically produced molecules as NAD+ enhancers in cell culture.
AB - Dysregulation of nicotinamide adenine dinucleotide (NAD+) homeostasis by increased activity of NAD+ consumers or reduced NAD+ biosynthesis plays an important role in the onset of prevalent, often age-related, diseases, such as diabetes, neuropathies or nephropathies. To counteract such dysregulation, NAD+ replenishment strategies can be used. Among these, administration of vitamin B3 derivatives (NAD+ precursors) has garnered attention in recent years. However, the high market price of these compounds and their limited availability, pose important limitations to their use in nutritional or biomedical applications. To overcome these limitations, we have designed an enzymatic method for the synthesis and purification of (1) the oxidized NAD+ precursors nicotinamide mononucleotide (NMN) and nicotinamide riboside (NR), (2) their reduced forms NMNH and NRH, and (3) their deaminated forms nicotinic acid mononucleotide (NaMN) and nicotinic acid riboside (NaR). Starting from NAD+ or NADH as substrates, we use a combination of three highly overexpressed soluble recombinant enzymes; (a) a NAD+ pyrophosphatase, (b) an NMN deamidase, and (c) a 5′-nucleotidase, to produce these six precursors. Finally, we validate the activity of the enzymatically produced molecules as NAD+ enhancers in cell culture.
KW - Enzymatic synthesis
KW - NAD precursor
KW - NaMN
UR - http://www.scopus.com/inward/record.url?scp=85147604145&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.foodres.2023.112560
DO - https://doi.org/10.1016/j.foodres.2023.112560
M3 - Article
C2 - 36869544
SN - 0963-9969
VL - 165
JO - Food Research International
JF - Food Research International
M1 - 112560
ER -