CHAPTER 7

The Chemistry and Biochemistry

of Niacin (B3)
ASDRUBAL AGUILERA-MÉNDEZ,a CYNTHIA
FERNÁNDEZ-LAINEZ,b ISABEL IBARRA-GONZÁLEZc
AND CRISTINA FERNANDEZ-MEJIA*c
a
Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria,
CP 58030 Morelia, Michoacan, Mexico; b Laboratorio de Errores Innatos del
Metabolismo y Tamiz, Instituto Nacional de Pediatrı́a, Av. del Iman #1, 9th
floor, Mexico City, CP 04530, Mexico; c Unidad de Genética de la Nutrición,
Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de
México/Instituto Nacional de Pediatrı́a, Av. del Iman #1, 4th floor, Mexico
City, CP 04530, Mexico
*Email: crisfern@biomedicas.unam.mx

7.1 Introduction
Niacin, also known as vitamin B3, nicotinic acid or vitamin PP, is a water-
soluble B-complex vitamin (Table 7.1). This vitamin is the generic descriptor
for two vitamers: niacin and niacinamide. In the research literature the terms
nicotinic acid/nicotinamide are most commonly used, while in medical practice
niacin/niacinamide are preferred. The vitamin is obtained from the diet in the
form of nicotinic acid, nicotinamide and tryptophan, which are transformed to
nicotinamide adenine dinucleotides, NAD and NADP. These compounds
participate in cellular oxidation–reduction reactions that are critical for
energy production. NAD and NADP also participate in a wide variety of

Food and Nutritional Components in Focus No. 4

B Vitamins and Folate: Chemistry, Analysis, Function and Effects
Edited by Victor R. Preedy
r The Royal Society of Chemistry 2013
Published by the Royal Society of Chemistry, www.rsc.org

108
The Chemistry and Biochemistry of Niacin (B3) 109
Table 7.1 The chemistry of niacin and niacinamide. Vitamin B3 vitamers:
niacin and niacinamide are composed of a pyrimidine ring bound
to a carboxylic group or to a carboxamide group.

IUPAC nomenclaturea Pyridine 3-carboxylic acid

CAS numberb 59-67-6
Molecular weighta 122.101 g/mol
NIACIN

IUPAC nomenclaturea Pyridine 3-carboxamide

CAS numberb 98-92-0
Molecular weighta 122.124 g/mol
NIACINAMIDE
a
Source: PubChem.
b
Source: ChemIndustry.

ADP-ribosylation reactions such as DNA repair, calcium mobilization and

deacetylation (Kirkland 2009). In addition, at pharmacological concentrations,
niacin is an effective agent for the treatment of dislipidemias and atherosclerosis
(Prousky et al. 2011). Furthermore, evidence exists that niacin ameliorates
acute migraine, chronic tension-type headaches, depression and schizophrenia
(Prousky et al. 2011). This chapter focuses on chemical and biochemical aspects
of the vitamin.

7.2 Niacin Chemistry

Nicotinic acid and nicotinamide (Table 7.1) are colourless crystalline sub-
stances; each is insoluble or only sparingly soluble in organic solvents. Nico-
tinic acid is slightly soluble in water and ethanol; nicotinamide is very soluble in
water and moderately soluble in ethanol. The two compounds have similar
absorption spectra in water, with absorption maxima at 262 nm.
Nicotinic acid is zwitterionic in nature; at high pH it is negatively
charged at the carboxylic function, while at low pH it is positively charged at
the pyridinyl nitrogen. Thus it is considered an amphoteric molecule because
it forms salts with acids as well as bases (Mullangi and Srinivas 2011)
(Figure 7.1).
Both nicotinic acid and nicotinamide are very stable in dry form (Gonçalves
et al. 2011); in solution nicotinamide is hydrolysed by acids and bases to yield
nicotinic acid. The standard molar enthalpy of formation of nicotinic acid
involved in protonation/deprotonation equilibrium of its three species at infi-
nite dilution is Df H1m (HN1 C5H4COOH NH2O, aq) ¼ (328.2  1.2) kJ/mol,
Df H1m (HN1 C5H4COO NH2O, aq) ¼ (325.0  1.2) kJ/mol, and Df H1m
(N1 C5H4COO NH2O, aq) ¼ (313.7  1.2) kJ/mol (Gonçalves et al. 2011).
110 Chapter 7

Figure 7.1 Nicotinic acid is an amphiprotic system composed of three species in

equilibrium. Nicotinic acid at high pH is negatively charged at the car-
boxylic function, while at low pH it is positively charged at the pyridinyl
nitrogen.

7.3 Niacin Daily Requirement, Food Sources and

Niacin Deficiency
The vitamin is obtained from the diet in the form of nicotinic acid, nicotina-
mide, NAD/NADP and tryptophan. The Recommended Dietary Allowance
(RDA) for adults is 16 mg/day of niacin equivalents for men and 14 mg/day for
women (Food and Nutrition Board 1998).
Niacin in mature cereal grains, particularly in corn, is largely bound and is
poorly available; alkali treatment of the grain increases the percentage absor-
bed. Meat and fish have the scarce free form of niacin and niacinamide but
contain high levels of NAD/ NADP, which are available as niacinamide after
digestion (Prousky et al. 2011). Fortification of flour and cereal products adds
up to 20 mg of the free form of niacin per serving to items such as breakfast
cereals (Food and Nutrition Board 1998).
Subclinical niacin deficiency is still present in developing countries and frank
deficiency occurs in cancer patients, alcoholics and anorexia nervosa (Kirkland
2007; Prousky et al. 2011).
Niacin deficiency, named Pellagra, is characterized by diarrhoea, dermatitis,
dementia and death, which usually appear in this order. The clinical expressions
of pellagra are diverse (Prousky et al. 2011). Diagnosis was, and still is, difficult
due to the unpredictable appearance of the different signs and symptoms in
individual patients (Prousky et al. 2011). Pellagra can be divided into primary
and secondary forms. Primary pellagra results from inadequate niacin and/or
tryptophan in the diet. Secondary pellagra occurs when other diseases or fac-
tors affect niacin requirements.

7.4 Factors and Diseases Affecting Niacin Requirement

The requirement for preformed niacin tends to be lower with higher tryptophan
intakes, while the requirement for preformed niacin is increased by factors that
reduce the conversion of tryptophan to niacin. These factors include low
tryptophan intake and inadequate iron, riboflavin or vitamin B6 status, which
participate in the conversion of tryptophan to niacin (Food and Nutrition
Board 1998). Other cases of reduced conversion of tryptophan to niacin are
The Chemistry and Biochemistry of Niacin (B3) 111
Hartnup disease, an autosomal recessive trait that interferes with the absorp-
tion of tryptophan, and carcinoid syndrome in which the amino acid is pre-
ferentially oxidized to 5-hydroxytryptophan and serotonin. Prolonged
treatment with the drug isoniazid, which competes with pyridoxal 5 0 -phosphate
(a vitamin B6-derived coenzyme required in the tryptophan-to-niacin pathway),
also reduces the conversion of tryptophan to niacin. Oral contraceptives that
contain high doses of estrogen increase tryptophan conversion efficiency
(Braidman and Rose 1971).
Niacin requirements are also affected by diseases or conditions interfering
with niacin absorption and/or processing, such as prolonged diarrhoea, chronic
dialysis treatment, chronic colitis (particularly ulcerative colitis), cirrhosis of
the liver, tuberculosis of the gastrointestinal tract, malignant carcinoid tumour
and chronic alcoholism (Food and Nutrition Board 1998). Substantial indivi-
dual differences (about 30%) in the conversion efficiency of tryptophan to
niacin have been reported (Horwitt et al. 1981).

7.5 Metabolism
To date there is no consensus with respect to the NAD/NADP preferred
synthesis substrate. Feeding of rats with tryptophan, nicotinic acid or nicotin-
amide showed that the first resulted in the highest NAD concentration in liver,
suggesting that this is the favoured substrate at least in this organ. In tissues
that lack the complete de novo NAD biosynthesis pathway, nicotinamide is
thought to be chosen over nicotinic acid as the main precursor for NAD
biosynthesis (Houtkooper et al. 2010). Excess niacin is methylated in the liver
to N1-methyl-nicotinamide, which is excreted in the urine along with the 2- and
4-pyridone oxidation products of N1-methyl-nicotinamide (Mrochek et al. 1976).

7.5.1 Niacin de novo Synthesis from Tryptophan

The kynurenine–anthranilate pathway, which is part of tryptophan catabolism,
is involved in the liver in the conversion of tryptophan to nicotinamide
(Figure 7.2). The first step is catalysed by either tryptophan 2,3 dioxygenase or
indolamine-pyrrole 2-3 dioxygenase—iron porphyrin metalloproteins which
oxidize the pyrrole moiety of L-tryptophan and represent the rate-limiting
enzyme of the kynurenine pathway (Thomas and Stocker 1999). In mammals,
tryptophan 2, 3 dioxygenase is the major enzyme contributing to NAD bio-
synthesis in the liver, while in extrahepatic tissues, indolamine-pyrrole 2,3
dioxygenase plays an important role. Dioxygenases catalyse cleavage of the
indole ring with incorporation of two atoms of molecular oxygen, forming
N-formylkynurenine. Tryptophan 2,3 dioxygenase is positively induced in liver
by cortisol and by tryptophan (Oxenkrug 2010). Tryptophan dioxygenase is
feedback-inhibited by nicotinic acid derivatives, including NADPH (Michal
1999). In a second step, the hydrolytic removal of the formyl group of N-
formylkynurenine, catalysed by arylformamidase, produces L-kynurenine,
which is then hydroxylated by kynurenine-3 monooxygenase resulting in
112 Chapter 7

Figure 7.2
The Chemistry and Biochemistry of Niacin (B3) 113
3-L-hydroxykynurenine. Hydroxylation requires molecular oxygen in a NADPH-
dependent reaction. Further transformation of 3-L-hydroxykynurenine in 3-
hydroxyanthranylate and alanine is catalysed by kynureninase, a pyridoxal
phosphate enzyme. A deficiency of vitamin B6 results in reduced niacin synthesis
due to the failure to catabolyse these kynurenine derivatives. Kynureninase is
positively regulated by tryptophan. In a subsequent step, the action of 3-
hydroxyanthranilate 3,4-dioxygenase causes ring opening. The resulting semi-
aldehyde undergoes a spontaneous condensation and rearrangement to form
quinolinate, which is the initial compound for NAD and NADP biosynthesis
(Figure 7.2).

7.5.2 NAD Biosynthesis

7.5.2.1 From Tryptophan
Quinolinate decarboxylation and conversion to nicotinic acid mononucleotide
is catalysed by quinolinate phosphoribosyltransferase, a rate-limiting enzyme
in the conversion of tryptophan to NAD; the reaction requires Mg21 and
is negatively regulated by nicotinamide. Next the transfer of adenylate from
ATP by an intermediate of nicotinamide/nicotinate-mononucleotide-adenyl-
transferases isoenzymes (NMNAT, see below) yields nicotinic acid adenine

Figure 7.2 Nicotinamide, nicotinic acid and tryptophan are utilized through distinct
metabolic pathways to form NAD and NADP. Tryptophan is converted
to NAD in the eight-step de novo pathway. First, tryptophan is converted
to N-formylkynurenine by the gene products of either indoleamine
dioxygenase (INDO) or tryptophan dioxygenase (TDO2). From N-L-
formylkynurenine, arylformamidase (AFMID) forms kynurenine.
Kynurenine is then used as a substrate of kynurenine monooxygenase
(KMO) and forms 3-hydroxykynurenine. Kynureninase (KYNU) then
forms 3-hydroxyanthranilate, which is converted to 2-amino-3-carboxy-
mucoaldehyde by 3-hydroxyanthranilate dioxygenase (HAAO). The
semialdehyde undergoes a spontaneous condensation and rearrangement
to form quinolinate, which is converted to nicotinic acid mononucleotide
by quinolinate phosphoribosyltransferase (QPRT). Nicotinic acid
mononucleotide is then adenylylated by NMNAT1-3 genes to form
nicotinic acid adenine dinucleotide, which is converted to NAD by
glutamine-dependent NAD synthetase (NADSYN1). Nicotinic acid is
utilized in the three-step Preiss–Handler pathway. Nicotinic acid phos-
phoribosyltransferase (NAPRT1) forms nicotinic acid mononucleotide
by addition of the 5-phosphoribose. In two steps shared with the
de novo pathway, nicotinic acid mononucleotide is then converted to
nicotinic acid adenine dinucleotide and finally to NAD via activity of
nicotinamide/nicotinate-mononucleotide-adenyltransferases isoenzymes:
NMNAT1-3 and NAD synthase-1 NADSYN1. Nicotinamide, from the
diet and from ADP transfer reactions, is converted to NAD via nicoti-
namide phosphoribosyltransferase (NAMPT/PBEF1), which catalyses the
addition of a phosphoribose moiety onto nicotinamide to form nicotina-
mide mononucleotide, this latter is subsequently converted to NAD by
NMNAT1-3.
114 Chapter 7
dinucleotide. NAD negatively regulates this step. Finally, nicotinic acid ade-
nine dinucleotide is transformed by the catalytic action of NAD synthase-1 in
the amide, NAD (Figure 7.2).

7.5.2.2 Salvage Pathways from Niacin and Niacinamide

Humans use both nicotinic acid and nicotinamide to synthesize NAD coenzymes,
but utilize different salvage pathways to achieve this. An enzyme in common
between the pathways is the adenylation enzyme, NMNAT. This enzyme has three
isoforms in humans (NMNAT-1, NMNAT-2 and NMNAT-3). NMNAT-1 is the
most proficient catalyst as determined by catalytic velocity (Vmax) and efficiency
(Vmax/Km); the enzyme is localized in the nuclei. Isoenzymes NMNAT-2 and -3 are
localized in the Golgi and in mitochondria, respectively (Sauve 2008). All isoforms
exhibit dual specificity for both nicotinic acid mononucleotide and nicotinamide
mononucleotide (Sauve 2008) (Figure 7.2).

7.5.2.2.1 Salvage Pathway from Niacin. In humans, nicotinic acid is con-

verted to nicotinic acid mononucleotide by the catalytic action of nicotin
phosphorybosyltransferase-1. By the action of NMNAT(s) and NAD syn-
thase-1, this compound is then transformed to NAD (Figure 7.2).

7.5.2.2.2 Salvage Pathway from Nicotinamide. Although an activity that

converted nicotinamide to NAD independently of nicotinic acid had been
indicated for some time, the enzyme was not identified until recently
(Rongvaux et al. 2002). Nicotinamide is coupled directly to phosphorybosyl
pyrophosphate to form nicotinamide mononucleotide by nicotinamide phos-
phoribosyltransferase, an enzyme that was previously identified as a cytokine
pre-B-cell colony-enhancing factor (PBEF) and controversially claimed as an
insulin-mimetic hormone visfatin (Imai 2009) (Figure 7.2).

7.5.3 NADP Biosynthesis

The generation of NADP is catalysed by NAD kinase, which transfers a
phosphate group from ATP onto the 2 0 -hydroxyl group of the ribose moiety of
NAD. A single mammalian NAD kinase has so far been identified; NAD
kinase activity is essential for cell survival (Agledal et al. 2010).

7.5.4 NAD Recycling

Humans have metabolic pathways that are able to recycle NAD. The main
pathway is catalysed by ADP-ribosyltransferases, which participate in non-
redox adenosine diphosphate (ADP)–ribose transfer reactions. These NAD-
consuming enzymes break down NAD to nicotinamide and ADP-ribosyl
product. Nicotinamide is then retransformed to NAD by the enzymatic action
The Chemistry and Biochemistry of Niacin (B3) 115
of nicotinamide phosphoribosyltransferase (NAMP/PBEF) and NMATs
(Figure 7.3).
Recently, a recycling pathway independent of nicotinamide was found to be
broadly conserved in bacteria, yeast and mammals (Bieganowski and Brenner
2004) The pathway leads from the metabolite nicotinamide riboside, the
dephosphorylated form of nicotinamide mononucleotide. A highly biologically
conserved nicotinamide riboside kinase is able to use nicotinamide riboside as a
substrate and can convert nicotinamide riboside to nicotinamide mono-
nucleotide (Bieganowski and Brenner 2004). Subsequently, nicotinamide
mononucleotide is converted to NAD by the catalytic action of NMNATs. In
humans, two isoforms of the kinase (Nrk1 and Nrk2) have been cloned,
although little is known about the biochemical properties and regulation of
these enzymes (Sauve 2008) (Figure 7.3).

7.6 NAD/NAPD Chemical and Structural Proprieties

NAD and NADP are composed of two nucleotides joined through their
phosphate groups by a phosphoanhydride bond (Table 7.2). NADP differs
from NAD in the presence of an additional phosphate group on the 2 0 position
of the ribose ring (Table 7.2). These coenzymes are white amorphous powders
that are hygroscopic and highly water-soluble.
NAD and NADP act as electron acceptors during the enzymatic removal of
hydrogen atoms from specific substrate molecules. One hydrogen atom from
the substrate is transferred as hydride ion to the nicotinamide portion of the
oxidized forms of these coenzymes to yield the reduced coenzymes NADH or
NADPH, respectively; the other hydrogen atom from the substrate becomes a
hydrogen ion (Figure 7.4). The reduced nucleotides absorb light at 340 nm but
the oxidized forms do not. This difference in absorption is used to assay
reactions involving these coenzymes.
Most dehydrogenases that use NAD or NADP bind the cofactor in a con-
served protein domain called the Rossmann fold. This structure results in a
relative loose binding that permits the coenzyme diffusion from one enzyme to
another.

7.7 NAD/NADP Metabolic Actions

7.7.1 Energy Metabolism and Oxidation Processes
NAD and NADP play a central role in energy metabolism, both being involved
in many cellular oxidation–reduction reactions. NAD functions in energy-
producing reactions involving the catabolism of innumerable metabolites.
NAD formed from oxidation reacts at the point of Complex I of the mito-
chondrial electron transport chain. Each mole of NADH consumed by the
mitochondria furnishes energy for the formation of three moles of ATP.
NADH is highly depleted in the cytosol. The phosphorylated dinucleotide,
 116

Figure 7.3 NAD recycling. Humans have two metabolic pathways that are able to recycle nicotinamide. NAD-consuming enzymes (ARTs,
PARPs, sirtuins) break down NAD to nicotinamide and ADP-ribosyl product. Nicotinamide by the enzymatic action of nico-
tinamide phosphoribosyltransferase (NAMP/PBEF) and nicotinamide/nicotinate-mononucleotide-adenyltransferases isoenzymes
Chapter 7

(NMAT1-3) is then retransformed to NAD. In a second pathway, nicotinamide riboside is phosphorylated by nicotinamide
riboside kinase (NRK 1,2) to nicotinamide mononucleotide. Subsequently, nicotinamide mononucleotide is converted to NAD by
the catalytic action of NMNATs.
The Chemistry and Biochemistry of Niacin (B3) 117
Table 7.2 The chemistry of NAD and NADP. Nicotin adenine dinucleotides:
NAD and NADP are composed of two nucleotides joined through
their phosphate groups by a phosphoanhydride bond.

IUPAC name [[(2R,3S,4R,5R)-5-(6-

aminopurin-9-yl)-3,4-
dihydroxyoxolan-2-
yl]methoxy-hydroxy-
phosphoryl][(2R,3S,4R,5R)-
5-(3-carbamoylpyridin-
1-ium-1-yl)-3,4-dihydroxy-
oxolan-2-yl]methyl hydro-
gen phosphate
CAS number 53-84-9
(chem. Industry)
Chemical formula C21H28N7O14P2 þ
(pubchem)
Molecular weight 664.433042 [g/mol]
(pubchem)

NAD+

IUPAC name [[(2R,3R,4R,5R)-5-(6-amino-

purin-9-yl)-3-hydroxy-
4-phosphonooxyoxolan-2-
yl]methoxy-hydroxyphos-
phoryl][(2R,3S,4R,5R)-5-
(3-carbamoylpyridin-1-
ium-1-yl)-3,4-dihydroxy-
oxolan-2-yl]methyl
hydrogen phosphate
CAS number 53-59-8
(chem. Industry)
Chemical formula C21H29N7O17P3 þ
(pubchem)
Molecular weight 744.412943 [g/mol]
(pubchem)

NADP+

NADP, functions in biosynthetic (anabolic) reactions such as the synthesis of

fatty acids cholesterol, bile acids, steroid hormones and the amino acids glu-
tamate and proline. NADPH þ H is also essential for the reduction of ribo-
nucleotides to deoxyribonucleotides and is thereby indirectly involved in DNA
synthesis.
118 Chapter 7

Figure 7.4 NAD/NADP redox reactions. NAD and NADP act as electron acceptors
during the enzymatic removal of hydrogen atoms from specific substrate
molecules. One hydrogen atom from the substrate is transferred as a
hydride ion to the nicotinamide portion of the oxidized forms of these
coenzymes to yield the reduced coenzymes NADH or NADPH, respec-
tively; the other hydrogen atom from the substrate becomes a hydrogen ion.

7.7.2 Protective Action of NADP

In addition to its biosynthetic role, NADPH is the unique provider of reducing
equivalents to maintain or regenerate the cellular detoxifying and antioxidant
defence systems. NADPH is an important cofactor for P450 enzymes that
The Chemistry and Biochemistry of Niacin (B3) 119
detoxify xenobiotics in reactions converting relatively insoluble organic com-
pounds to hydrophilic ones, thereby facilitating their breakdown and secretion.
In oxidative defence, NADPH acts as a terminal reductant for glutathione
reductase, which maintains reduced glutathione. In some cell types, NADPH is
required for the reactivation of catalase, when the enzyme is inactivated by
H2O2. In addition, NADPH serves as a substrate for NADPH oxidase, which
generates peroxides for release in oxidative burst processes of the immune
system (Agledal et al. 2010).

7.7.3 Non-redox Adenosine Diphosphate (ADP)-Ribose

Transfer Reactions
NAD/NADP have also shown to be required for important non-redox ade-
nosine diphosphate (ADP)-ribose transfer reactions involved in DNA repair,
calcium mobilization and deacetylation reactions (Lautier et al. 1993; Pollak
et al. 2007) (Figure 7.5).
Three classes of enzymes cleave the b-N-glycosylic bond of NAD to free
nicotinamide and catalyse the transfer of ADP-ribose in non-redox reactions
(Lautier et al. 1993). The first class consists of ADP ribose transferases: mono-
ADP-ribosyltransferases and poly-ADP-ribose polymerase, which catalyse
ADP-ribose transfer to proteins. The second class correspond to ADP-ribosyl
cyclases—enzymes that promote the formation of cyclic ADP-ribose, a com-
pound that mobilizes calcium from intracellular stores in many types of cells
(Pollak et al. 2007). The third class of NAD consuming enzymes consists of
sirtuins—proteins that possess either histone deacetylase or mono-ribosyl-
transferase activity.

7.7.3.1 Mono-ADP-ribosyltransferases
These enzymes catalyse the ADP-ribose moiety of NAD transfer to an acceptor
amino acid. Five mammalian ADP-ribosyltransferases have been cloned and
expression is restricted to tissues such as cardiac and skeletal muscle, leuko-
cytes, brain and testis. ADP-ribosyltransferases-1 and -2 are glycosylphos-
phatidylinositol (GPI)-anchored ectoenzymes. ADP-ribosyltransferase-5
appears not to be GPI-linked and may be secreted. In skeletal muscle and
lymphocytes, ADP-ribosyltransferases-1 modifies specific members of the
integrin family of adhesion molecules, suggesting that ADP-ribosylation affects
cell-matrix or cell-cell interactions (Okazaki and Moss 1999).

7.7.3.2 Poly-ADP-ribose Polymerases

These enzymes catalyse the attachment of ADP-ribose polymers to target
proteins and represent an elaborate protein modification. The most studied
human enzyme among 17 members of this family, poly-ADP-ribose poly-
merase-1 (PARP-1), is involved in DNA repair and apoptotic pathways.
PARP-1, a nuclear enzyme which detects DNA damage, binds to DNA single
 ADP-ribosylation
Poly (PARPs)
Mono
120

ADP ribosylated Poly-ADP

protein ribosylated protein
Sirtuins

NAD+
Deacetylated
protein

ADP-ribosyl
cyclases

Ca+2

Cyclic-ADP-Ribose
ER

Figure 7.5 Adenosine diphosphate (ADP)-ribosylation biochemical reactions. Mono-ADP-ribosyltransferases (ARTs) and poly-ADP-
ribose polymerases Poly (PARPs) catalyse the ADP-ribose moiety of NAD transfer to amino acid residues. ADP-ribosyl cyclases
generate cyclic ADP-ribose and 2-phospho-cyclic ADP-ribose from NAD and NADP, respectively. Both molecules trigger cyclic
Chapter 7

ADP-ribose cytosolic Ca21 elevation, presumably by activating the ryanodine receptor in the endoplasmic/sarcoplasmic reti-
culum (RER). SIRT1 catalyses a reaction that couples protein deacetylation to NAD hydrolysis.
The Chemistry and Biochemistry of Niacin (B3) 121
or double strand breaks, and then catalyses the transfer of many ADP-ribose
units from NAD to an acceptor protein and also to the enzyme itself. The
intensity of DNA damage determines cellular pathways: survival, apoptosis, or
necrosis. In the case of mild DNA damage, poly(ADP-ribosylation) enhances
DNA repair and thus cell survival. When the damage is beyond repair, PARP-1
facilitates apoptosis, preventing ATP depletion. Severe DNA damage leads to
PARP-1 over-activation, cellular energy depletion and necrotic cell death
(Surjana et al. 2010). Poly-ADP-ribosylation has also important functions in
telomere dynamics, transcriptional regulation, cell division and trafficking of
endosomal vesicles (Pollak et al. 2007).

7.7.3.3 ADP-ribosyl Cyclases

These enzymes generate cyclic ADP-ribose and 2-phospho-cyclic ADP-ribose
from NAD and NADP, respectively. Both molecules trigger cyclic ADP-ribose
cytosolic Ca21 elevation, presumably by activating the ryanodine receptor in
the endoplasmic/sarcoplasmic reticulum (Pollak et al. 2007). In addition to
cyclic ADP-ribose, niacin adenine dinucleotide phosphate, a metabolite of
NADP, can also mobilize Ca21 stores. The release mechanism and the stores on
which niacin adenine dinucleotide phosphate acts are from lysosomal Ca21
stores, which are independent of the stores of activated by cyclic ADP-ribose or
inositol 1,4,5-trisphosphate (Yamasaki et al. 2004).

7.7.3.4 Sirtuins
Sirtuins are a highly conserved family of protein deacetylases and ADP-
ribosyltransferases, whose distinguishing characteristic is a requirement for the
oxidized form of NAD (Lomb et al. 2010). The dependence of sirtuins upon
NAD plays a major role in connecting their enzymatic activity to the grade of
energy of the cell by means of the cellular NAD/NADH ratio. Seven mammalian
orthologs (SIRT1–7) have been described, with SIRT 1 the best studied. SIRT1
catalyses a reaction that couples lysine deacetylation to NAD hydrolysis. The
deacetylation reaction of sirtuins consists of two steps. In the first, sirtuins cleave
NAD and produce nicotinamide, and in the second step, the acetyl group is
transferred from the substrate to the ADP-ribose moiety of NAD to generate
O-acetyl-ADP ribose and the deacetylated substrate. Nicotinamide is a strong
inhibitor of SIRT1 deacetylase activity. Fasting has been found to increase
SIRT1 protein levels and activity. SIRT1 has also found to be activated in
response to oxidative stress, low glucose availability and endurance exercise
(Lomb et al. 2010). Importantly, SIRT-1 regulates the activity of several tran-
scription factors and cofactors by modulating their acetylation status, and by
this means links the metabolic state of the cell to transcriptional regulation.

7.8 Concluding Remarks

In this chapter we have summarized several interesting findings that extend the
actions of NAD/NADP from oxidoreductase cofactors to signalling molecules.
122 Chapter 7
Furthermore, novel data have unveiled the link between the cellular redox
status and the control of metabolism, cellular viability and transcriptional
events. Continued progress will be needed to understand NAD/NADP meta-
bolism and signalling in health and disease.

Summary Points
 Niacin is also known as vitamin B3, nicotinic acid or vitamin PP. It is the
generic descriptor for two vitamers: niacin and niacinamide.
 The vitamin is obtained from the diet in the form of nicotinic acid,
nicotinamide and tryptophan.
 Niacin is a precursor to NAD and NADP, coenzymes whose classical role
is to participate in redox reactions.
 In addition to their classical role as coenzymes, NAD and NADP parti-
cipate in a wide variety of ADP-ribosylation reactions, which are involved
in cell signalling and the control of many cell processes.
 ADP ribose transferases: mono- and poly-ADP-ribose polymerases cata-
lyse ADP-ribose transfer to proteins.
 ADP-ribosyl cyclases promote the formation of cyclic ADP-ribose, a
compound that mobilizes calcium from intracellular stores in many types
of cells.
 Sirtuins possess either histone deacetylase or mono-ribosyltransferase
activity. The dependence of sirtuins activity upon NAD plays a major role
in connecting their enzymatic activity to the grade of energy of the cell by
means of the cellular NAD/NADH ratio.

Key Facts about Niacin History

 American natives knew that corn needed to be processed to be nutritious
in a process known nixtamalization (alkaline treatment commonly used in
Mexico and Central America in the preparation of tortillas, tamales and
hominy) or ingested as immature ears. This knowledge was lost when
Christopher Columbus returned from the New World with samples of the
new seed.
 The use of corn without processing spread over time and gradually the
clinical signs of niacin deficiency became recognized as a disease.
 In 1771, Francesco Frapolli made the first reference to the disease as
pellagra, meaning ‘rough skin’ in Italian. He linked it with poverty and
subsistence on nutritionally marginal corn-based diets.
 In 1937, Conrad A. Elvehjem isolated the P-P (pellagra preventive) factor,
identified it as nicotinic acid (niacin).
 The term ‘niacin’ was coined to avoid the associations of nicotinic acid
with acids and nicotine, which shocked the public.
The Chemistry and Biochemistry of Niacin (B3) 123
Definitions of Words and Terms
ADP-ribosylation reactions: Reaction whereby an ADP-ribose moiety is linked
covalently to another compound.
Apoptosis: Programmed cell death.
De novo synthesis: Synthesis of complex molecules from simple molecules such
as sugars or amino acids, as opposed to their being recycled after partial
degradation.
DNA repair: A collection of processes by which a cell identifies and corrects
damage to the DNA molecules that encode its genome.
Ectoenzymes: Catalytic membrane proteins with their active sites outside the
cell.
Necrosis: Uncontrolled cell death that leads to lysis of cells and inflammatory
responses.
Nixtamalization: Refers to a process for the preparation of maize (corn) or
other grain in which the grain is soaked and cooked in an alkaline solution,
usually limewater, and hulled.
Orthologs: Orthologs, or orthologous genes, are genes in different species that
are similar to each other because they originated by vertical descent from a
single gene of the last common ancestor.
Recommended Dietary Allowances: These refer to the recommended daily levels
of nutrients to meet the needs of nearly all healthy individuals in a particular
age and gender group.
Redox: Oxidation–reduction.
Salvage pathway: A pathway that utilizes compounds formed in catabolism for
biosynthetic purposes, even though these compounds are not true inter-
mediates of the corresponding normal biosynthetic pathway.
Telomere: Repetitive DNA sequences at the end of a chromosome, which
protects the end of the chromosome from deterioration or from fusion with
neighbouring chromosomes.
Toxicokinetics: The description of what rate a chemical will enter the body and
what happens to it once it is in the body.
Vitamers: One or multiple related chemical compounds having a given vitamin
activity.
Xenobiotics: A compound that is foreign to an organism. It can also cover
substances that are present in much higher concentrations than are usual.
Zwitterionic: A molecule carrying both a positive and a negative charge.

List of Abbreviations
AFMID arylformamidase
ARTs mono ADP-ribosyltransferases
Df H1m standard molar enthalpy of formation
GPI glycosylphosphatidylinositol
HAAO 3-hydroxyanthranilate dioxygenase
124 Chapter 7
INDO indoleamine dioxygenase
IUPAC International Union of Pure and Applied Chemistry
kJ/mol kilo Joule per mol
KMO kynurenine monooxygenase
KYNU kynureninase
NAD nicotinamide adenine dinucleotide
NADP nicotinamide adenine diphosphate
NADSYN1 NAD þ synthetase-1
NAMP nicotinamide phosphoribosyltransferase
NMNAT nicotinamide/nicotinate mononucleotide adenyltransferases
NAPRT1 nicotinic acid phosphoribosyl transferase-1
NRK 1,2 nicotinamide ribiside kinase 1 and 2
PARP-1 poly-ADP-ribose polymerase-1
PBEF pre-B-cell colony-enhancing factor
QPRT quinolinate phosphoribosyltransferase
RDA Recommended Dietary Allowance
RER endoplasmic/sarcoplasmic reticulum
SIRT silent information regulator proteins
TDO2 tryptophan dioxygenase

Acknowledgments
The authors are grateful to Luis Rodriguez-Villela for illustrations. Their work
was supported by research grants from the Consejo Nacional de Ciencia y
Tecnologı́a 99294-M and the Dirección General de Asuntos del Personal
Académico, Universidad Nacional Autónoma de México IN214811. Asdrubal
Aguilera is recipient of the CONACyT scholarship number 91634 and
PROMEP, folio UMSNH-208.

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