151.

232 ~ Nutrition and metabolism

The B-group (part A)

2015
A/Prof Rozanne Kruger

B-group vitamins
learning objectives
To be able to describe their absorption, transport and
storage
To be able to describe important dietary sources of each
vitamin
To be able to discuss the metabolic functions of these
vitamins and the consequences of suboptimal and
deficient intakes
To know how to assess them in vivo
Relevant pages in
Thompson: chapters 8-12

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B vitamins names and functions

Thiamin
Riboflavin*
Niacin
Pyridoxine*
Cobalamin
Folate
Pantothenic acid
Biotin
Inositol

B1
B2
B3
B6
B12
B9
B5
B7
B8

Metabolism of energy yielding

intermediates via red/ox reactions
Transamination
Transmethylation*
Constituent of coenzyme A
Carboxylation using coenzyme A

Absorption sites

Pathways involving B-vitamins

A number of the
coenzymes involved
in important
metabolic pathways
require B vitamins
for their activity

PLP = Vit B6
NAD/NADP = Niacin (B3)
TPP = Thiamin (B1)
FMN/FAD = Riboflavin (B2)
CoA = Pantothenic acid
THF = Folate
Biotin, Vit B12

Thiamin discovery
First discovered as essential through
deficiency symptoms of a condition known
as beriberi, that occurred with use of
polished rice in late 19th and early 20th
centuries.
Thiamin is removed from cereals by
refining, as most of the thiamin is in the
husk of grains.
Polished rice is very low in thiamin
However, inward diffusion of water soluble
vitamins means that parboiled rice retains
most of its thiamin.

I cannot, I cannot
1870
Marked increase in
beriberi incidence in SE Asia3
Polished rice

Christiaan Eijkman
White rice can be poisonous!
(1896)3
The anti-beriberi factor

Gerrit Grijns
Suggested dietary deficiency
(1901)4

Thiamin
RDI
men: 1.2 mg/day
women: 1.1 mg/day
NRVS NHMRC 2006

Thiamin sources
Whole grains,
fortified, or enriched
grain products
Moderate amounts in
all nutritious food
Pork

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Absorption
Absorbed by an active saturable process
Mainly from upper intestine
Maximum absorbed from a single dose
2.5mg
At intakes >5mg per day a second passive
absorption process operates
Alcohol inhibits the active transport
process

Storage and excretion

Very little free thiamine stored in the tissues
Most (90%) is present as active co-enzyme,
thiamin pyrophosphate (thiamin diphosphate)
10% in brain (thiamin triphosphate) role
unclear
Circulates in the blood bound to albumin
Excreted intact in urine, with renal
conservation during pregnancy

Measuring vitamin status functional markers

Enzyme activation co-efficients
Method
Obtain blood sample and measure enzyme
activity
Add a known dose of the vitamin coenzyme
Re -measure the enzyme activity again
The enzyme function after the addition of the
vitamin coenzyme is divided by the pre-addition
test result to give the activation co-efficient
an increase in ETK (erythrocyte transketolase) of >20%
is indicative to risk of thiamin deficiency, an increase of
15% is considered normal.

ETK-AC > 1.2= deficiency

The ETK-Ac increases with greater deficiency

Vit B1~ Thiamin

Assists in energy metabolism as a coenzyme pyrovate acetyl coA (TPP Coenzyme)
Also role on nerve cell membranes

Functions of Thiamin
Central role in energy-yielding metabolism,
particularly metabolism of CARBOHYDRATE
Thiamin pyrophosphate (TPP) is the coenzyme
for 3 oxidative carboxylation reactions:
Pyruvate dehydrogenase in carbohydrate (CHO)
metabolism (required for conversion of pyruvate to
acetyl- coenzyme A) (pyruvate & lactate accumulate)
Alpha keto glutarate dehydrogenase in the citric acid
cycle
Branched chain keto acid dehydrogenase involved in
amino acid metabolism (leucine, isoleusine, valine)

Functions of Thiamin
Thiamin pyrophosphate is also the coenzyme
for transketolase, in the pentose phosphate
pathway of CHO metabolism (alternative
pathway of glucose metabolism)
Thiamine triphosphate has a function in nerve
conduction

Causes of deficiency
Diets high in polished rice (in W world most
thiamin comes from fortified breads/cereals
Malnourished alcoholics have marked reductions
in thiamin
Outbreaks of thiamin deficiency disease are often
seasonal due to a general shortage of foods
Some foods contain anti-thiamin factors e.g. shell
fish and ferns (thiaminase)
Breast fed infants are at risk if mother has low
thiamin intake (breast milk is not a good source of
thiamin)

Deficiency: Beriberi
2 major manifestations of thiamin deficiency:
Cardiovascular disease Wet beriberi
Neurological disease Dry beriberi

Symptoms:
Wet beriberi (acute) swelling (oedema), increased heart
rate (tachycardia), lung congestion, congestive heart
failure due to accumulation of pyruvate & lactate prompt
response to treatment
Dry beriberi (chronic) pain, tingling or loss of sensation in
hands and feet (peripheral neuropathy), muscle wasting
with loss of function of lower extremities (inability to lift foot
= foot drop), absent ankle jerk, brain damage and death.

Wet beriberi

Dry beriberi

Symptoms
Vision changes
Double vision
Eye movement abnormalities
(nystagmus)
Eyelid drooping
Loss of muscle coordination
Unsteady, uncoordinated
walking
Loss of memory, can be profound
Inability to form new memories
Confabulation (making up stories)
Hallucinations
Staggering

Deficiency: Beriberi
In western word main cause of thiamin
deficiency- alcoholism
Wernicke encephalopathy alcohol related
brain damage language problems, walking
difficulty, unusual eye movement
Korsakoff Syndrome (Korsakoffs psychosis)
amnesia, inability to learn or retain new
memories, confabulation

Riboflavin
Other names: Vitamin B2
2006 RDI
Men: 1.3 mg/day
Women: 1.1 mg/day

Chief functions in the body

Part of coenzymes FMN (flavin
mononucleotide) and FAD (flavin adenine
dinucleotide) used in energy metabolism.
Participate in the flavoproteins in the
oxidation chain in the mitochondria

Riboflavin
Sources

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Riboflavin Vitamin B2
Principal dietary sources:
Milk products, poultry, meat, fish.
Smaller amounts in vegetables and cereals, but may
contribute a high proportion of requirements if these are
a major part of the diet.

Dietary riboflavin occurs as FAD and FMN

protein bound.
Easily destroyed by ultraviolet light, irradiation
Opaque containers NOT SEE-THROUGH

Heat stable

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Recommended daily intake for

Riboflavin.
The difference between an intake that is
inadequate and may lead to deficiency
(0.45-0.55 mg/day) and an intake that will
saturate tissue stores (~1.1 mg/day) is
very small.
This is because the storage capacity is
very limited.

Absorption of Riboflavin
Dietary FAD and FMN are released from
protein by stomach acid and hydrolysed to
free riboflavin or FMN bound to albumin in
the upper intestine.
Riboflavin is absorbed by a sodium
dependent, saturable process, and
converted to FMN in the intestinal cells.

Transport, storage and excretion.

Transported bound to albumin.
Stored mainly as FAD, some as FMN, little
as free riboflavin.
Most storage is in liver, also in muscle
Excreted in the urine
Some enters the enterohepatic circulation

Measurement of Riboflavin
Measurement of activity of red cell
glutathione reductase, with and without
addition of FAD.
Erythrocyte glutathione reductase activation
co-efficient (EGRAC)

Ratio of activity with FAD of 1.2 or higher

indicates a risk of deficiency.

Riboflavin chief functions

Part of coenzymes:
FMN (flavin mononucleotide) and
FAD (flavin adenine dinucleotide)
used in energy metabolism - oxidation chain in
the mitochondria (picks up 2 hydrogens from
TCA cycle)

Riboflavin chief functions

The enzymes that require FMN or FAD as
cofactors are termed flavoproteins.
Several flavoproteins also contain metal
ions and are termed metalloflavoproteins.
Both classes of enzymes are involved in a
wide range of redox reactions, e.g.
succinate dehydrogenase and
xanthine oxidase,
NADH dehydrogenase.

Riboflavin

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Riboflavin chief functions

FMN and FAD electron acceptors participate as
cofactors in many oxidation-reduction reactions
e.g. in the catabolism of glucose, fatty acids,
ketone bodies and amino acids.
coenzyme that transfers energy from one
compound to another
if AA are used for energy, necessary for
deamination
essential for growth and tissue repair
If thiamin lacking, riboflavin likely lacking too

Deficiency disease: Ariboflavinosis

Deficiency symptoms
Inflamed eyelids and sensitivity to light,
reddening of cornea (photophobia)
Sore throat
Cracks and redness at corners of mouth
(angular stomatitis)(cheilosis)[perlche]
Painful, smooth, purplish red
(magenta) tongue (glossitis)
Inflammation characterized by skin
lesions covered with greasy scales (seborrhea)
Growth retardation
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Deficiency disease: Ariboflavinosis

Deficiencies
NZ inadequate intakes low but more likely in
Pacific children or females (11-18yrs; Mori;
with food security concerns)
Vegans or others who dont consume animal
products must ensure ample servings of dark
green leafy veges
Toxicity symptoms: none reported
Highly unlikely in NZ

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Glossitis

glossitis

angular stomatitis

Cheilosis

Capillary infiltration of the cornea

Angular palbebritis

Dyssebacia
sebaceous plugs

Niacin (Nicotinic Acid)

Deficiency disease is called pellagra
(rough skin shark skin / sandpaper).
First recognised during the 19th Century in
Europe and Southern USA maize diets,
and in India with millet diets.
Animal protein reversed the deficiency
symptoms, and eventually the important
factor in protein was shown to be
tryptophan.

Niacin
Other names
Nicotinic acid
Nicotinamide
Niacinamide
Vitamin B3

Precursor: dietary tryptophan

2006 RDI
Men: 16 mg NE/day
Women: 14 mg NE/day

Upper level for adults: 35 mg/day

Copyright 2005 Wadsworth Group, a division of Thomson Learning

Niacin
Chief functions in the body
Part of coenzymes NAD (nicotinamide adenine
dinucleotide) and NADP (its phosphate form)
used in energy metabolism

Significant sources
Milk, eggs, meat, poultry, fish
Whole-grain and enriched breads +cereals
Nuts
All protein-containing foods
Also converted from tryptophan
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Niacin
Sources

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Copyright 2005 Wadsworth Group, a division of Thomson Learning

Conversion of tryptophan to niacin.

Note that vitamin B6 is required for this
process.
A high leucine : isoleucine ratio may
reduce tryptophan availability and cause
pellagra, particularly if vitamin B6 is also
low.

Dietary tryptophan: a source of

niacin
For normal adults in nitrogen balance, all
required niacin can be obtained by
synthesis from dietary tryptophan:
60 mg of tryptophan are required to
synthesize 1 mg of niacin
Also, synthesis of niacin from tryptophan
requires vitamins B1, B2 and B6 which would
be limiting in themselves on a marginal diet.
EXAMPLE: Protein intake 80 g/day ;
Tryptophan ~ 1000 mg (12.6 mg/g protein) ;
Yields ~ 17 mg niacin using the conversion
factor: 60 mg diet. tryptophan = 1 mg niacin.

Absorption, transport and excretion

Nicotinic acid and nicotinamide absorbed by
sodium-dependent saturable transport and by
passive diffusion.
Circulate in plasma, enter tissues by diffusion.
Converted to co-enzyme forms in various tissues
Many catabolites of niacin have been identified,
but only two appear in the urine.

Functions
Niacin is converted to NAD+ and NADP.
Essential for the function of all enzymecatalysed reactions that involve NAD+
(catabolism) and NADP (anabolism).
Vital for glycolysis, fatty acid metabolism, tissue
respiration, detoxification reactions and many
other pathways.
necessary for getting energy from the macronutrients
synthesis of fatty acids and cholesterol

supports health of
skin
nervous system
digestive system

1900s pellagra caused 87000 deaths due to low

protein diet with high corn content
HOW?
low protein diet / high corn diet high leucine in
corn interferes with tryptophane absorption
B3 in Corn too complex to absorb (niacytin)
bound to complex CHO & small peptides

Deficiency: Pellagra
Pellagra - the 4 Ds (black tongue in dogs)
Dermatitis is most characteristic, and occurs in
skin areas exposed to sunlight.
Diarrhoea not always seen.
Dementia - mental changes may include
confusion, loss of memory, disorientation and
various psychoses.
Death
Mouth signs similar to riboflavin deficiency.

Deficiency: Pellagra

Pellagra
Pellis=skin
Agra=rough

Diarrhoea
Dermatitis*
Dementia
Death
Skin darkens and flakes
Bilateral & Symmetrical
Occurs only on parts exposed to sunlight

Hartnups disease
Pellagra is seen in Hartnups disease.
This is a disorder of amino acid
absorption, in which several amino acids,
including tryptophan, are poorly absorbed.

Assessment of niacin deficiency

No satisfactory blood measure
Measurement of urinary metabolites is
used to assess risk of deficiency.
Urinary concentrations of metabolites Nmethylnicotinamide and N-methyl-2pyridone-5-carboxamide are decreased
in deficiency.

Toxicity symptoms
Painful flush, hives, and rash
(niacin flush)
at intakes>200mg/d
Excessive sweating
Blurred vision
Liver damage, impaired glucose
tolerance

Pharmacological use
Nicotinic acid is the most effective broad
spectrum lipid drug doses 1-3g
Reduces total cholesterol and increases
HDL cholesterol concentrations
Decreases circulating free fatty acids by
inhibiting lipolysis in adipose tissue
Limitations?
possible liver damage and diabetes

Vitamin B6
Three forms (interconvertible):
Pyridoxine (alcohol form) - plants
Pyridoxal (aldehyde form) - animals
Pyridoxamine (amine form) - animals
All three forms can undergo phosphorylation.
The active form is pyridoxal phosphate .
2006 RDI
Adults (19-50 years): 1.3 mg/day

Upper level for adults: 50 mg/day

Vitamin
B6
Sources
Meats, fish,
poultry
Potatoes,
legumes, non
citrus fruits
Fortified
cereals
Liver
Soy products
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Functions
Part of coenzymes PLP (pyridoxal phosphate) and PMP
(pyridoxamine phosphate) used in amino acid
(homocysteine) and CHO metabolism (DNA & RNA)
Fatty acid metabolism
formation of essential FA (linoleic acid arachidonic acid)
Synthesis & turnover of cholesterol

Main function in protein metabolism transamination and

deamination of proteins
NE amino acids (serotonin, threonine)
Synthesis & metabolism of
tryptophan (trp niacin)
Methionine to taurine,
GABA

Formation of cross-linkages in elastin formation

Maintenance of cellular immunity - production of antibodies

Helps make RBCs form precursor of porphyrin ring of
haeme (part of haemoglobin)

Functions
Release glycogen from the liver and muscle
as glycogen phosphate.
Forms part of the phosphorilase enzyme
(glycogen glc-1-phosphate)(gluconeogenesis)

Form sfingolipids necessary for development

of myelin sheaths in nerve cells
CNS metabolism & ability to transmit nerve
impulses (synthesis of neurotransmitters
(serotonin, norepinephrine, taurine,
dopamine), tryptophan serotonin)

Physiological roles of PLPdependent enzymes:

Transamination reactions involved in
oxidative metabolism of amino acids.
Decarboxylation reactions giving amines.
Glycogen breakdown glycogen
phosphorylase.
Synthesis of niacin from tryptophan
Haeme synthesis

Role of B6 in regulation of hormone

action
Terminates (turns off) hormone action by
releasing hormone receptor complexes
Modulates the actions of the steroid
hormones
oestrogens, androgens, progesterone, retinol,
thyroid hormones and calcitriol (active form of
vitamin D)

Absorption, metabolism and

excretion
Most B6 in foods as PLP bound to
enzymes
B6 dephosphorylated before absorption,
taken up by diffusion, then rephosphorylated for incorporation into
enzymes where required.
PLP not incorporated into enzymes is
dephosphorylated, and oxidised to 4pyridoxic acid in the liver for excretion.

Deficiency: anaemia
Severe deficiency is rare:
leads to seizures,
may produce anaemia lack of haeme.

Deficiency may occur with:

malabsorption problems,
dialysis for chronic renal failure,
chronic alcoholism.

Marginal deficiency may be more common, and

may predispose to atherosclerosis and cancer of
the breast, uterus and prostate.
Plasma [B6] tends to decrease with age.

Deficiency
Neurotransmitters diminish
Abnormal Tryptophan synthesis products
accumulate in brain
Symptoms

Depression
Confusion
Abnormal brain wave patterns
Convulsion

Alcohol loss of B6
Broken down = acetaldehyde dislodges PLP
coenzyme from enzyme excreted

Oral Contraceptives depletes PLP, however

controversy exists

Dermatitis greasy, flaky skin

Glossitis

Microcytic anemia

Microcytic anemia

Drugs may cause B6 deficiency

The following drugs:
Isoniazid (tuberculosis treatment)
Penicillamine (rheumatoid arthritis,
Wilsons disease)
Benserazide, carbidopa (Parkinsons
disease)
Oral contraceptives
May cause B6 depletion, which in turn also
cause niacin (nicotinic acid) depletion.

Assessment of B6 status
Best methods are:
Plasma concentration of PLP
Erythrocyte transaminase activation.
? Methionine load
(incomplete metabolism of methionine can result
in increased homocysteine)
Non-specific measure = Tryptophan load tests
If person has deficiency, the aa trp get trapped
and the intermediate kynurenine is shunted to
xanthurenic acid rather than niacin (test
presence in urine).

Requirements for B6
High protein in diet increases losses, so
requirements are linked to dietary protein.
Intakes of 15-16 g/g protein are
recommended.
RNI 1.5 -1.6 mg/day at an average protein
intake of 100 g/day.

Toxicity of B6
Very high doses (2-7 g/day) may cause
neurological damage / problems.
Supplements 200-2000mg have been
used for treating depression, carpal tunnel
syndrome and premenstrual syndrome,
but there is little scientific evidence to
support the effectiveness.
May be toxic.
Safe upper limit 100 mg/day.

Pantothenic Acid
Panthos = everywhere
As a coenzyme
takes part in the metabolism of carbohydrates
and fats
It is essential for the oxidation of pyruvic acid
More than 100 steps in synthesis of lipids ,
neurotransmitters , steroid hormones and
haemoglobin

Part of Co Enzymes (Coenzyme A!)

Present in TCA cycle

Recommendation = 5mg/d adults

Pantothenic Acid
2006 adequate intake (AI)
Adults: Men: 6 mg/day
Women: 4mg/day

Significant sources
Widespread in foods
Organ meats, mushrooms, avocados,
broccoli, whole grains

Easily destroyed by food processing

Pantothenic Acid
A dimethyl derivative of butyric acid which is
linked to b-alanine.
Essential component of Coenzyme A (CoA)
lipid and CHO metabolism,
fatty acid synthesis,
steroid hormone synthesis,
gluconeogenesis.

Key element in energy production (Krebs cycle)

used for

energy release from the macronutrients

synthesis of fat
formation of haeme
synthesis of cholesterol and neurotransmitters

Absorption, metabolism & excretion

CoA in foods hydrolysed by intestinal
phoshatases to produce pantothenic acid.
Absorbed as pantothenic acid, via active
transport through a saturable, sodiumdependent carrier.
CoA is resynthesized in liver cells.
Pantothenic acid is excreted in the urine.

Assessment of status
Pantothenic acid can be measured in urine
and blood microbiologically or radioimmunologically.
Normal blood levels > 100mg/dl
Normal urinary excretion 1-15 mg/day
Urinary values are used as a sensitive
indicator of dietary intake.

Pantothenic Acid
Deficiency symptoms
Rare observations made from clinically
induced states
Vomiting, nausea, stomach cramps
Insomnia, fatigue, depression, irritability,
restlessness, apathy
Hypoglycemia, increased sensitivity to insulin
Burning feet syndrome

Toxicity symptoms: none reported

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Biotin

Chief functions in the body

Part of a coenzyme that carries activated carbon
dioxide. The role is critical in the TCA cycle:
biotin delivers a carbon to 3-carbon pyruvate,
thus replenishing oxalo-acetate,
the 4-carbon compound needed to combine with
acetyl CoA to keep the TCA cycle turning.

Also participates in:

gluconeogenesis,
fatty acid synthesis, and
the breakdown of certain fatty acids and amino acids

Recent research had uncovered a role in gene

expression

Dietary sources and availability

Biotin is present in liver, egg yolk, soy
flour, cereals and yeast.
Fruit and meat are poor sources.
Generally plant sources have higher
content than animal sources
Availability is governed by binders in food:
in raw egg white, and in wheat biotin is
present in an unavailable bound form.

Dietary sources and availability

Significant sources
Widespread in foods
Organ meats, egg yolks, soybeans, fish,
whole grains
The protein avidin found in raw egg whites
binds biotin and prevents absorption
Avidin is destroyed by heating

Also produced by GI bacteria large

intestine (may not contribute much to biotin
absorbed)

2006 adequate intake (AI)

very small quantities
Adults: 30 g/day

Digestion and absorption

Protein-bound biotin in the diet is digested to
give:
biocytin,
biotin with a lysine residue covalently bound, or
biotinyl-lysyl peptides.

These are hydrolysed by a specific enzyme

called biotinidase.
Free biotin is taken up by sodium-dependent
active transport.
Biocytin is only slowly taken up by passive
diffusion.

Metabolism and excretion

Absorbed biotin is
converted to biotinyl-5-adenylate (in a
reaction with ATP) ;
then incorporated into the apocarboxylase
forms of the four carboxylases for which it
serves as a cofactor.

Biotin and metabolites are excreted in

urine at a rate of about 6-50 mg/day.

Metabolic role of biotin

The only known metabolic role of biotin is
as a coenzyme for 4 carboxylase enzymes:
Acetyl CoA carboxylase (fatty acid synthesis)
Pyruvate carboxylase (oxalo-acetate synthesis
for the TCA cycle, gluconeogenesis)
3-methylcrotonyl CoA carboxylase (catabolism
of ketogenic aa leucine)
Propionyl CoA carboxylase (catabolism of oddchain fatty acids and some amino acids)

Biotin Deficiency
Human biotin deficiency is rare; occurs
only with:
very unusual diets
egg white injury,

incorrect total parenteral nutrition (TPN),

severe malnutrition, or
chronic use of some anticonvulsant drugs
(phenytoin, primidone, phenobarbital,
carbamazepine).

Biotin Deficiency - symptoms

Neurological impairment:
Depression, lethargy, hallucinations, numbness or
tingling sensation in the arms and legs

Skin rash (seborrhoeic dermatitis) red scaly

rash around the eyes, nose and mouth (due to
altered fatty acid metabolism), glossitis
Hair loss (alopecia)
General symptoms include fatigue, nausea,
anorexia, muscle pains, anaemia, and elevated
serum cholesterol (hypercholesterolaemia).

Assessment of deficiency
Assay of biotin-dependent carboxylase activity in
lymphocytes has been used to monitor biotin
status in malnutrition.
Low activity of the carboxylases has been
reported in studies on children with marasmus or
kwashiorkor.
Supplementation with biotin increased the
activity of the carboxylases.
Biotin assayed in whole blood or urine by
microbiological assay (normal whole blood 0.22
0.75 mg /ml)

Summary so far
B1
(TPP)

Pyrovate

Biotin

B3
(NAD/NADP)
B2
(FAD/FMN)