Vitamins and Minerals

Domingo J Piero, PhD Nutrition, Food Studies, and Public Health

Outline
Essential nutrients Vitamins and minerals Clinical signs of deficiencies B vitamins in alcoholism

Essential Nutrients for Humans

Water Amino acids: His, Ile, Leu, Lys, Met, Phe, Thr, Trp, Val Fatty acids: linoleic, -linolenic Vitamins: A, D, E, K, ascorbic acid, thiamin, riboflavin, niacin, pyridoxine, pantothenic acid, folate, biotin, vitamin B12 Minerals: Ca, P, Mg, Fe Trace minerals: Zn, Cu, Mn, I, Se, Mo, Cr Electrolytes: Na, K, Cl Ultratrace elements: As, B, F, Ni, Si, V Energy sources

Criteria for Essential Nutrient

The substance is not synthesized de novo in the body The substance is required for growth and/or maintenance of health Lack of exogenous intake results in characteristic signs of a deficiency disease The signs of deficiency are prevented only by the nutrient or a specific precursor of it

Conditional Essentiality
Substances not ordinarily required in the diet for healthy humans, but which are essential for specific populations or under certain circumstances
Pathologic states (e.g., liver cirrhosis, requires Cys, Tyr and taurine; severe trauma requires Gln) Immature infants (appear to require Cys and Tyr, elongated derivatives of w-6 and w-3 fatty acids, and carnitine) Genetic defects (carnitine synthesis)

Conditional Essentiality
Three tests proposed to establish a condition of conditional essentiality:
Decline in the plasma level of the nutrient into the subnormal range Appearance of chemical, structural, or functional abnormalities Correction of both of 1 and 2 by a dietary supplement of the nutrient

Conditional Essentiality
Grey area e.g., the following vitamins could be considered conditionally essential:
Niacin when there is a relative tryptophan deficiency, since it can be made from tryptophan Vitamin D when exposure to sunlight is low, since is made on the surface of the skin when exposed to sunlight

Vitamins
A large group of potent organic compounds necessary in minute amounts in the diet Divided based on their solubility characteristics
Water-soluble vitamins usually function as coenzymes in the metabolism of fats, proteins and carbohydrates Fat-soluble vitamins have a variety of function in some cases similar to hormones

Minerals
First subdivided based on the magnitude of need: macrominerals and microminerals Function as cofactors in intermediary metabolism, and the formation and maintenance of the bodys structure

General functions of some vitamins

A B2 B3 B6 retinoic acid retinal riboflavin niacin pyridoxine pyridoxamine pyridoxal cobalamin ascorbic acid cholecalciferol gene expression phototransduction redox, respiration redox aa metabolism, glycogenolysis

B12 C D

1C metabolism hydroxylation bone remodeling gene expression

E
K Biotin Choline Folate

tocopherols
phytylmenaquinone multiprenylmenaquinone

antioxidant
coagulation bone remodeling gluconeogenesis, tca, fa, aa pl 1C metabolism tca, fa and cholesterol

Panthotenic acid

Functional classification
Bone health: Vitamin A, vitamin K, calcium, phosphorus, magnesium Nutritional anemias: Fe, Zn, Cu, B6, Folate, B12, riboflavin Antioxidant capacity: Selenium, vitamin E, vitamin C, vitamin A, Fe, Cu

Pharmacological Uses of Micronutrients

In high doses, some micronutrients have pharmacologic effects, different from their nutritional function. For example:
Large doses of nicotinic acid lower serum cholesterol I.V. infusions of Mg are used to treat preeclampsia and myocardial infarction

Dietary Reference Intakes (DRIs)

Set of nutrient recommendations by Institute of Medicine (IOM) of the U.S. National Academy of Sciences
EAR (Estimated Average Requirement): daily intake level estimated to meet the requirement of half the healthy individuals in a particular life stage and gender group RDA (Recommended Dietary Allowance): average daily intake level sufficient to meet the requirements for nearly all (97-98%) healthy individuals in a particular life stage and gender group RDA = EAR + 2SD if SD available If SD not known, usually assume coefficient of variation (SD/mean) = 10%, so RDA = EAR + 20%

Dietary Reference Intakes (DRIs)

AI (Adequate Intake): used when an EAR and RDA cannot be determined; based on observed or experimentally determined approximations or estimates of intakes by a group of healthy people assumed to be adequate UL (Tolerable Upper Intake Level): highest average longterm intake likely to pose no risk of adverse effects to almost all individuals in the general population

DRIs in Perspective
EAR
1.0

RDA AI?

UL

Risk of Adverse Events

Risk of Inadequacy

0.5

Observed Level of Intake

Nutrient Requirements Affected by:

Age, sex (EARs, RDAs/AIs based on these) Genetics Drug-nutrient interactions Nutrient-nutrient interactions Amount of nutrient precursor (e.g., tryptophan intake affects niacin requirement; 60 mg tryp = 1 mg niacin)

Assessing Individual Nutritional Status

Dietary and medical history Serum/plasma, urine, or red or white blood cells nutrient levels
Serum/plasma levels indicate amount transported, not necessarily stored -- however, the body doesnt store water-soluble vitamins, though it does store fat-soluble vitamins and many minerals Urinary levels indicate recent intake and use RBCs good for mid- to long-term status; WBC more complicated

Assessing Individual Nutritional Status

Assess status of specific tissues or organs to determine if specific micronutrient requirements are not met. Examples:
Rickets, poor growth and development of bones due to deficiency of vitamin D and/or calcium Anemia, due to deficiency of iron, certain other minerals or certain B vitamins

General Issues
Signs and symptoms non-specific Function of nutrient not understood at molecular level Lack of adequate assessment tools Clinical nutrition physical assessment

Vitamin A (and carotenoids)

Functions:
Normal vision Growth and reproduction Cell proliferation, differentiation, and modulation of apoptosis Immune system integrity

Food sources:
Liver Fish oil Eggs Fortified milk or other foods Red, yellow, orange, and dark green veggies (carotenoids)

Vitamin D (the sunshine vitamin)

Functions:
Calcium homeostasis Antiproliferative, prodifferentiation, proapoptotic agent as well as inhibitor of cell migration Plays role in immunity Calciotropic versus noncalciotropic functions

Sources:
Sunlight (10 15 min 2x a week) Fish (with bones) Fortified milk and other foods

Vitamin E
Functions:
Antioxidant*

Sources:
Vegetable oils Foods made from oil (salad dressing, margarine) Nuts Seeds Wheat germ Green, leafy veggies

CH3 HO CH3 O CH3 CH3 CH3 CH3 CH3

CH3

-tocopherol

O2H2O2 Lipid-OO Lipid-OOH

O2

Vitamin E redox cycle

O2Dehydroascorbate

Ascorbate
CH3

GSSG GSH

O
O CH3 CH3

CH3

CH3

CH3 CH3

CH3

-tocopheroxyl

Vitamin K Function

COO-

-OOC

COOCH

CH2
CH2

Vitamin K

CO2

CH2

-glutamyl carboxylase -carboxy glutamic acid (Gla) in peptide

Glu in peptide

Carboxylation of glutamyl residues

Vitamin K
Functions:
Blood coagulation: 4 procoagulant (prothrombin, factors VII, IX, and X) and 3 anti-coagulant factors (proteins C, S, and Z) Bone health Cell replication and transformation, cell survival, cell signaling

Sources:
Body can produce on its own (from bacteria in intestines) Green, leafy veggies Some fruits, other veggies, and nuts

Thiamin (B1)
Functions:
Energy production Metabolism of branchedchain amino acids Pentose phosphate shunt

Sources:
Whole-grain and enriched grain products Pork Lentils Fortified cereals

Riboflavin (B2)
Functions:
Energy production Niacin synthesis from tryptophan Oxidant/antioxidant balance

Sources:
Milk Enriched cereals Eggs Almonds Green, leafy veggies

Niacin
Functions:
Metabolism of sugars/fatty acids Non-redox functions: Deacetylases, ribosyl cyclases, ribosyl tranferases and oolyADP-ribose polymerases (PARPs)

Sources:
Foods high in protein typically (poultry, fish, beef, peanut butter, legumes) Enriched and fortified grains

Pyridoxine (B6)
Functions:
Amino acid metabolism Neurotransmitter synthesis Glycogen metabolism Heme synthesis

Sources:
Animal protein Whole grains and fortified cereals Bananas Nuts Legumes

Folate (folic acid)

Functions:
1-carbon metabolism
DNA, RNA expression Homocysteine/methionine metabolism Serine/glycine metabolism Phospholipids Catecholamines Other methylation reactions

Sources:
Fortified and enriched grains and breakfast cereals Orange juice Legumes Green, leafy veggies Peanuts Avocados

Vitamin B12 (cobalamin)

Functions:
Metabolism of amino acids and fatty acids Synthesis of hemoglobin Cofactor for methionine synthase

Sources:
Shellfish Liver Animal products Eggs Milk, other dairy

Biotin
Functions:
Energy production Proteins, carbs, and fats metabolism Fixation of CO2

Sources:
Wide variety of foods Eggs Liver Wheat germ Peanuts Cottage cheese Whole grain bread

Pantothenic Acid
Component of CoA Sources: Found in almost all foods Helps the body use Meat, poultry, fish proteins, fat, and carbs Whole grain cereals from food
Legumes Milk Fruits, veggies

Vitamin C
Functions:
Antioxidant Posttranslational hydroxylation of proline and lysine in the formation of collagen Collagen gene expression CNS: neurotransmitter synthesis NE and serotonin) myelin synthesis Iron absorption Aldosterone and corticosteroids synthesis Vasodilatory and anticlotting effects through PGA synthesis

Vitamin C
Sources:
Black currants Broccoli, Brussels sprouts, cauliflower Citrus fruits Green leafy vegetables Potatoes

What are minerals?

Regulate body processes Give structure to things in the body No calories (energy) Cannot be destroyed by heat

Categories of minerals
Major minerals
Calcium Phosphorus Magnesium Electrolytes (sodium, chloride, potassium)

Trace minerals
Chromium Copper Flouride Iodine Iron Manganese Selenium Zinc

Calcium
Bone building Muscle contraction Heart rate Nerve function Helps blood clot

Phosphorus
Generates energy Regulate energy metabolism Component of bones, teeth Part of DNA, RNA (cell growth, repair) Almost all foods, especially protein-rich foods, contain phosphorus

Magnesium
Part of 300 enzymes (regulates body functions) Maintains cells in nerves, muscles Component of bones Best sources are legumes, nuts, and whole grains

Iron
Part of hemoglobin, carries oxygen Brain development Healthy immune system Sources:
Animals (heme) vs. plants (non-heme) Better absorbed from heme Consume vitamin C with non-heme Fortified cereals, beans, eggs, etc.

Deficiencies

Eyes
Normal Clinical Findings Suspect Deficiency Other Causes

Bright, clear, shiny, smooth cornea, membrane pink and moist

Pale conjunctiva
Night blindness Bitots spots Xerosis Redness and fissures in corners of eyelids

Fe
Vitamin A Vitamin A Vitamin A Riboflavin, pyridoxine Thiamin, phosphorus

Nonnutritional anemias
Hereditary eye diseases Aging, allergies

Normal eye movement Ophthalmoplegia to follow objects

Brain lesion*

Hair
Normal Shiny, firm, not easily plucked Normal appearing or thick Clinical Findings Suspect Deficiency Flag sign; easily plucked, no pain. Sparse Protein (kwashiorkor/ marasmus) Protein, biotin, zinc Other Causes Overprocessing of hair Alopecia from aging, chemotherapy, endocrine disorders, medication, etc.

Normal-appearing hair shaft and emergence from skin

Corkscrew hairs and unemerged coiled hairs

Vitamin C

Nails
Normal Uniform, rounded, smooth Clinical Findings Transverse ridging Suspect Deficiency Protein, Zinc? Normal if only in toenails Other Causes

Koilonychia Fe (thinning, flattening, concave, spoon-shaped) Brittle, soft Mg?

Skin (1)
Normal Uniform color; smooth, healthy appearance Clinical Findings Scaling or nasolabial seborrhea Suspect Deficiency Vitamin A, zinc, EFAs, riboflavin, pyridoxine Other Causes Hypervitaminosis A

Petechiae, esp. perifollicular

Ecchymosis

Vitamin C

Abnormal blood clotting, severe fever, flea bites

Anticoagulant therapy, injury, thrombocytopenia, hypervitaminosis E

Vitamin C, vitamin K

Skin (2)
Normal Uniform color; smooth, healthy appearance Clinical Findings Follicular hyperkeratosis Pigmentation, desquamation of sunexposed areas Cellophane appearance Yellow pigmentation, esp. palms of hands while sclera remains white Suspect Deficiency Vitamin A, vitamin C Niacin Other Causes

Protein

Aging process Excess carotene intake

Skin (3)
Normal Uniform color; smooth, healthy appearance Clinical Findings Suspect Deficiency Other Causes Medication, esp. steroids Poor skin care, DM

Body edema, moon Protein, face thiamin Poor wound healing, decubitus ulcers Pallor, fatigue Protein, vitamin C, zinc, kwashiorkor Fe

Blood loss

Oral (1)
Normal Lips smooth, no sores Clinical Findings Cheilosis Angular stomatitis Suspect Deficiency Riboflavin, pyridoxine, niacin Riboflavin, niacin, folate, vitamin B12, protein, Fe Other Causes Excessive salivation due to ill-fitting dentures

Tongue red Atrophic lingual without swelling, papillae normal surface

Glossitis (magenta, Riboflavin, niacin, scarlet, raw folate, vitamin B12, tongue) pyridoxine

Oral (2)
Normal Clinical Findings Suspect Deficiency Zinc Other Causes Medications (antineoplastic, sulfonylureas) Fluorosis Bulimia? Poor oral hygine, periodontal disease Vitamin C

Normal taste and smell Hypogeusia Hyposmia Normal gums and teeth Mottled enamel on teeth Eroded enamel Cavities, missing teeth, retracted gums Swollen, bleeding, retracted gums

Neurologic
Normal Psychological stability Clinical Findings Dementia Memory loss, confabulation, disorientation Normal reflexes Foot and wrist drop and sensations Peripheral neuropathy weakness, paresthesias Ataxia, and decreased tendon reflexes Suspect Deficiency Niacin, vitamin B12 Thiamin Other Causes Disease or agerelated, multiple causes (aluminum toxicity, increase serum calcium) Excess pyridoxine

Thiamin Thiamin, pyridoxine, vitamin B12

Alcoholism and B vitamins

Effects
Alcoholism places individuals at increased risk for vitamin B deficiency because of:
decreased intake decreased absorption/reabsorption impaired utilization

Thiamin
The active form of thiamin - thiamin diphosphate (TDP) works as a coenzyme in the following important reactions: Transketolase reaction The transfer of a 2C fragment from alpha-keto sugars to aldose acceptors in the pentose-phosphate shunt catalyzed by transketolase

Transketolase catalyzes two critical reactions in the pentose phosphate pathway

Functions of the pentose phosphate pathway:
1. To generate reducing equivalents in the form of NADPH, for reductive biosynthesis reactions within cells 2. To provide the cell with ribose-5-phosphate (R5P) for the synthesis of the nucleotides and nucleic acids 3. It can operate to metabolize dietary pentose sugars derived from the digestion of nucleic acids as well as to rearrange the carbon skeletons of dietary carbohydrates into glycolytic/gluconeogenic intermediates (Enzymes that function primarily in the reductive direction utilize the NADP+/NADPH cofactor pair as opposed to oxidative enzymes that utilize the NAD+/NADH cofactor pair)

TDP in the pentose phosphate pathway

Transketolase decreases early in thiamin deficiency; measuring its activity in RBCs has been used to assess thiamin nutritional status TK

Thiamin (TDP) TK

Thiamin
Oxidative decarboxylation of -keto acids The conversion of pyruvate to acetyl-CoA catalyzed by pyruvate dehydrogenase complex. The conversion of -ketoglutarate to succinyl-CoA in the TCA cycle catalyzed by alpha-ketoglutarate dehydrogenase. Rate limiting. The conversion of branched-chain alpha-keto acids to acyl-CoA's catalyzed by branched-chain alpha-ketoacid dehydrogenase.

Metabolism of BCAAs Val, Leu and Ile undergo reversible transamination to their corresponding -keto acids (by BCAA aminotransferase)
Val Leu Ile

+ -KG

Glu +

-ketovalerate -ketoisocaproate -keto--methylvalerate

The keto acids, then go through an irreversible decarboxylation of the carboxyl group by the branched chain ketoacid (BCKA) dehydrogenase to liberate CO2 The BCAAs are the only essential amino acids to undergo transamination; branched-chain amino acids and their ketoacids are toxic in excess (maple syrup urine disease) but have to be conserved for protein synthesis

Oxidative decarboxylation of -keto acids

Pyruvate dehydrogenase, -ketoglutarate dehydrogenase, and branched chain ketoacid (BCKA) dehydrogenase are mitochondrial enzyme complexes Catalyze the decarboxylation of pyruvate, -ketoglutarate, and branched-chain keto acids to form acetyl-coenzyme A, succinyl-coenzyme A, and derivatives of branched chain keto acids (acyl-CoAs), respectively, all of which play critical roles in the production of energy from food
In addition to the thiamin coenzyme (TDP), each dehydrogenase complex requires CoA, NAD, FAD, and lipoic acid

Glucose

TDP-dependent enzymes
Ribose-5-P
Xylulose-5P

G6P Pentose shunt

Glyceraldehyde-3-P Pyruvate Lactate

TK Sedoheptulose-7P

Leu

PDHC

Pyruvate
mitochondria
Ile
BCKADH

Acetyl-CoA OXA Citrate

Propionyl CoA Bt Methylmalonyl CoA B12

-KG Succinyl CoA

-KGDH

Glu

Val

MMA

GABA

Non-coenzyme function of thiamin

Less understood than TDP coenzyme role TTP concentrated in nerve and muscle cells Activates membrane ion channels, possibly by phosphorylating them Impaired formation of TTP may play role in neurological symptoms of severe thiamin deficiency

Evaluation of Thiamin Status Subjects history: dietary assessment, demographic data, medical history, family history, and psychological history should be recorded. Laboratory tests: urinary thiamin excretion blood TDP levels CSF thiamin concentration blood pyruvate, lactate, and -KG levels erythrocyte transketolase activity (ETKA); and TDP effect on ETKA:

Normal: 0-15% increase with TDP Marginal deficiency: 15-25% increase with TDP Deficiency: > 25% increase with TDP

Deficiency Affects the cardiovascular, muscular, nervous and gastrointestinal systems. Cardiac failure, muscle weakness, peripheral and central neuropathy, and gastrointestinal malfunction. The precise biochemical defects have not been established, but thiamin may as well play three major roles at the cellular level: Energy metabolism: related to the oxidative decarboxylation of -keto acids, the inhibition of which leads to a failure in ATP synthesis Synthesis mechanisms: reflected by the importance of the transketolase reaction in the formation of NADPH and pentose Neurotransmitters and nerve conduction: changes in GABA and glutamate

Beriberi: Clinical Signs

Clinical signs vary with age and the

organ systems involved Infantile Adult

Dry
Wet Wernicke encephalopathy and Wernicke-

Korsakoff Syndrome (sometimes referred to as cerebral beriberi)

Infantile beriberi
Most common between 2-6 mo Cardiac (acute fulminating) beriberi: acute attack

that includes a loud piercing cry, cyanosis, dyspnea, vomiting, tachycardia, and cardiomegaly, with death occurring within a few hour of the onset unless thiamin is administered Aphonic beriberi: the tone of the childs cry varies from hoarseness to complete aphonia Pseudomeningitic beriberi: infants exhibit vomiting, nystagmus, purposeless movements of the extremities, and convulsion accompanied by a normal cerebrospinal fluid

Adult Beriberi
Dry beriberi (paralytic or nervous) Main feature: peripheral neuropathy
Early on: "burning feet syndrome" may occur Other symptoms: exaggerated reflexes, diminished sensation

and weakness in the legs and arms Muscle pain/tenderness and difficulty rising from a squatting position also observed Seizures may be seen in severe deficiency

Wet beriberi (cardiac) In addition to neurologic symptoms, wet beriberi is characterized by cardiovascular manifestations, which include rapid heart rate, cardiomegaly, edema, difficulty breathing, and ultimately congestive heart failure

Beri beri

Cerebral beriberi
Wernicke-Korsakoff Syndrome = spectrum with 2

separate sets of symptoms, one of which tends to start when the other subsides
Wernicke's encephalopathy common in alcoholics, but

can also appear in people with HIV/AIDS, GI disease, or glucose IV without adequate B1 supplementation Involves damage to multiple nerves in both the central and peripheral nervous system Abnormal eye movements (e.g., nystagmus, ophtalmoplegia) Stance and gait abnormalities Abnormalities in mental function (apathy, memory problems) Peripheral neuropathy in 80% of patients

Cerebral beriberi
Korsakoff syndrome, or Korsakoff psychosis Involves impairment of memory (retrograde

as well as inability to learn) - patients often attempt to hide their poor memory by confabulating (making up stories about experiences or situations) Impairment of conceptual functions and decreased spontaneity and initiative. IV thiamin to WKS patients generally improves eye symptoms, but effects on motor coordination and memory may be less, depending on how long symptoms were present (nerve damage)

Related issues
Wernicke encephalopathy is underdiagnosed

doctors look for the classical triad: ophthalmoplegia, ataxia, confusion

Thiamin deficiency should be suspected in

grossly impaired nutritional status (especial attention to alcoholism, GI diseases, HIVAIDS, persistent vomiting)
Thiamin should be administered parenterally

as soon as possible
Essential before glucose solutions or

parenteral nutrition

Treatment
Beriberi patients should receive thiamin administration

as soon as possible
Usual dose = 50 - 100 mg/d IV or IM for 1-2 weeks,

followed by 10 mg/d orally until recovery

Change dietary habits / stop drinking alcohol Rationale for high dosage:
Replenish thiamin stores Stimulate TDP-dependent reactions maximally Improve cardiovascular disease

Riboflavin
Antioxidant function
Glutathione reductase is an FAD-dependent

enzyme that participates in redox cycle of glutathione, which protects organisms from reactive oxygen species, such as hydroperoxides

72

Glutathione Redox Cycle

73

Functions: Antioxidant
Xanthine oxidase, an FAD-dependent

enzyme, catalyzes oxidation of hypoxanthine to xanthine and further to uric acid, one of the most effective water-soluble antioxidants in the blood Riboflavin deficiency can result in decreased xanthine oxidase activity, reducing blood uric acid levels

74

75 Schematic diagram of the purine degradation pathway Pacher et al. Pharmacol Rev 58(1):87-114, 2006

Functions: Nutrient Interactions

B-complex vitamins B6, folate, B12, niacin
Conversion of vitamin B6 to its coenzyme form,

pyridoxal 5'-phosphate (PLP), requires the FMNdependent enzyme pyridoxine 5'-phosphate oxidase Synthesis of NAD and NADP from tryptophan requires the FAD-dependent enzyme kynurenine monooxygenase
Severe riboflavin deficiency can decrease conversion of tryptophan

to NAD and NADP, increasing risk of niacin deficiency

Methylene tetrahydrofolate reductase (MTHFR), an

FAD-dependent enzyme, plays important role in maintaining this specific form of folate (Me-THF), which is required to form methionine from homocysteine
76

Nutrient Interactions: Riboflavin, Folate, and Vitamin B12

77

Functions: Interaction with Iron

Riboflavin deficiency alters iron metabolism
Mechanism not clear, but may involve impaired iron absorption,

increased intestinal loss of iron, impaired Fe mobilization from ferritin and/or impaired iron utilization for the synthesis of hemoglobin

In humans, improving riboflavin nutritional

status has been found to increase circulating hemoglobin levels Correction of riboflavin deficiency in individuals who are both riboflavin deficient and iron deficient improves the response of irondeficiency anemia to iron therapy

78

Assessment
Urinary excretion of the vitamin in fasting,

random, or 24-hour specimens or by load return tests

Erythrocyte riboflavin concentration:

<27 nmol (10 g)/dL = deficiency

Erythrocyte glutathione reductase activity

coefficient, EGRac (before and after FAD addition) <1.2 acceptable, >1.4 deficient
79

Deficiency (Ariboflavinosis)
Rarely found in isolation; usually occurs in combination

with deficiencies of other water-soluble vitamins Symptoms include

sore throat redness (hyperemia) and swelling (edema) of the lining

of the mouth and throat cracks or sores on the outsides of the lips (cheilosis) and at the corners of the mouth (angular stomatitis) inflammation and redness of the tongue (magenta tongue) a moist, scaly skin inflammation (seborrheic dermatitis) corneal vascularization normochromic normocytic anemia with reticulopenia, leukopenia, and thombocytopenia peripheral neuropathy (hyperesthesia, and decreased sensitivity)
80

81

82

83

84

85

Biotin: fixing CO2

Biotin serves as a covalently bound coenzyme

for four important enzymes known as carboxylases These carboxylases catalyze an essential metabolic reaction: the incorporation of bicarbonate as a carboxyl group into a substrate
In all 4 carboxylases, biotin functions as a

coenzyme or prosthetic group that serves as a carrier for CO2 in a multistep reaction
86

The four carboxylases are:

Acetyl-CoA carboxylase (ACC) catalyzes the binding

of bicarbonate to acetyl-CoA to form malonyl-CoA. Malonyl-CoA is required for the synthesis of fatty acids. Cytosolic and mitochondrial.
Pyruvate carboxylase (PC) is a mitochondrial enzyme

critical in gluconeogenesis. Catalyzes the incorporation of bicarbonate into pyruvate to form oxaloacetate.
Methylcrotonyl-CoA carboxylase (MCC) catalyzes an

essential step in the metabolism of leucine. Located in mitochondria.

Propionyl-CoA carboxylase (PCC), participates in

essential steps in the metabolism of amino acids, cholesterol, and odd-chain fatty acids. Also mitochondrial.
87

FUNCTIONS OF BIOTIN

isoleucine 3-hydroxypropionate methionine methylcitrate, propionylglycine threonine odd-chain fatty acid valine Leucine propionyl-CoA

3-hydroxyisovalerate* 3-methylcrotonylglycine

Propionyl-CoA Carboxylase
d-methylmalonyl-CoA succinyl-CoA

3-methylcrotonyl-CoA

Methylcrotonyl-CoA Carboxylase
3-methylglutaconyl-CoA fatty acid synthesis, elongation FA oxidation malonyl-CoA

glucose

TCA cycle

oxaloacetate

pyruvate

acetyl-CoA

Pyruvate Carboxylase
lactate

Acetyl-CoA Carboxylase

88

Deficiency Very rare, the human requirement for dietary biotin has been demonstrated in two different situations:
prolonged intravenous feeding without biotin

supplementation, and
consumption of raw egg white for a prolonged period

(many weeks to years).

Long-term anticonvulsant therapy can lead to

biotin depletion to a level severe enough to interfere with amino acid metabolism
Other causes of depletion: pregnancy?,

dialysis, alcoholism.

89

Vitamin B12 Cofactor for methionine synthase: Methylcobalamin is a cofactor and folate is a substrate for the function of methionine synthase; enzyme required for the synthesis Met from homocysteine Met is required for the synthesis of Sadenosylmethionine (SAM), a methyl group donor used in many methylation reactions (DNA and RNA) Inadequate function of methionine synthase can lead to an accumulation of homocysteine, which has been associated with increased risk of cardiovascular diseases

One-carbon metabolism and transsulfuration pathway

Ser Gly B6 SHMT Gly B6 GLDC 5,10-CH2THF

Ser
Thymidine and purines CTH

THF: tetrahydrofolate; 5,10-CH2THF: 5,10methylene THF; 5-CH3THF: 5-methyl THF; SAM: S-adenosylmethionine; SAH: Sadenosylhomocysteine; MS: methionine synthase; BHMT: betaine-homocysteine methyltransferase; MAT: methionine adenosyltransferase; AK: adenosine kinase; ADA: adenosine deaminase; SAHH: SAH hydrolase; CBS: cystathionine beta synthase; CTH: cystathione gamma lyase; SHMT: serine hydroxymethyl transferase; GLDC: glycine decarboxylase

FUNCTION 2 Cofactor for L-methylmalonyl-CoA mutase: 5-Deoxyadenosylcobalamin is required by the enzyme L-methylmalonyl-CoA mutase that catalyzes the conversion of Lmethylmalonyl-CoA to succinyl-CoA Important in the production of energy from fats and proteins Succinyl CoA is also required for the synthesis of Hb

isoleucine methionine threonine valine propionyl-CoA

Leucine 3-methylcrotonyl-CoA

Propionyl-CoA Carboxylase
MMCoA mutase

Methylmalonic acid

d-methylmalonyl-CoA

Methylcrotonyl-CoA Carboxylase
3-methylglutaconyl-CoA fatty acid synthesis, elongation FA oxidation

succinyl-CoA glucose TCA cycle

oxaloacetate

pyruvate

acetyl-CoA

malonyl-CoA

Pyruvate Carboxylase

Acetyl-CoA Carboxylase

Diet Salivary glands

Protein-B12

R
IF HCl R TC I-B12 IF R-B12 IF R B12 IF-B12 TC II i r t o R-B12
Pancreatic proteases

Parietal cells Chief cells

Pepsin B12

Ileal IF-B12-Receptor (cubilin)

Portal circulation TC II-B12

B12

Pyrimidines synthesis

Purines synthesis

Lipid peroxyl radical LOO

LOOH

Vit Ered

VIT Eox

reduced products Vit Cox Vit Cred

Fe3+ (ferric) Cu2+ (cupric) hydroxyl radical (OH), superoxide radical anion (O2-)

Glutathione peroxidase (Se)

Glutathionered (GSH) +H2O2 Glutathioneox (GSSG)

Glutathione reductase (FAD)

Antioxidants working together

NADP+

NADPH, H+ Pentose phosphate pathway

Glucose-6-P

Ribulose-5-P

Questions?

B6: From diet to cell

Functions
The coenzyme PLP plays a vital role in the function of >100 enzymes that catalyze essential chemical reactions in the human body:
Decarboxylation of amino acids to produce amines important in neurotransmitter synthesis Transamination of amino acids to keto acids fuel Phosphorolytic cleavage of glycogen to G1P One-carbon metabolism and transsulfuration Synthesis of:
Carbohydrates, sphingolipids, amino acids, heme, neurotransmitters

Amino acid metabolism

Transamination: ALT and AST among others Transsulfuration (met to cys): cytathione synthase and cystathionase Selenoamino acid metabolism: release of Se from selenohomocysteine by the enzymes selenocysteine and -lyase Tryptophan-niacin conversion
Kynureninase catalyzes an important step; in B6 deficiency, the pathway is impaired and kynurenic and xanthurenic acids accumulate in urine

Histamine synthesis (histidine decarboxylase)

Glucose metabolism
Transaminations (in gluconeogenesis) Glycogen utilization
coenzyme for glycogen phosphorylase in the phosphorylation of glucose to G-1P. This function accounts for more than half the activity of B6 in the body

CNS function
Neurotransmitter synthesis
Serotonin
Tryptophan decarboxylase

Dopamine (and epi- and norepinephrine)

DOPA decarboxylase (aromatic L-amino acid decarboxylase)

GABA
L-glutamic acid decarboxylase

Energy production Neuronal function

Hemoglobin synthesis and function

Synthesis of heme
-aminolevulinate synthase condensates succinyl CoA and glycine to form -aminolevulinate, precursor of the porphyrin ring microcytic, hypochromic anemia

B6 binds to the and chains enhancing O2 binding and inhibiting sickling in sickle-cell Hb

Lipid metabolism
Synthesis of sphingolipids (serine palmitoyl transferase) Biosynthesis of carnitine (3-hydroxymethyl-lysine hydrolase). This reaction also needs iron.

Gene expression
Elevated intracellular B6 alters steroid hormone receptor-induced enzymes
Decreased response to glucocorticoids

Elevated expression of albumin mRNA in B6 deficiency Known to suppress mRNA levels for a a number of other proteins (apolipoprotein A-1, glyceraldehyde-3-phosphate dehydrogenase) and to decrease mRNA levels for others (RNA polymerase I and II)