2 6. PYRIDOXINE (vitamin B6) Pyridoxal (PL)Pyridoxamine (PM)Pyridoxine orPyridoxol (PN)Each of these forms can be phosphorylated at position 5 to form: PLP, PMP, and PNP.
3 Active form Pyridoxal phosphate (PLP) Active functional form is pyridoxal phosphate (PLP) and pyridoxamine phosphate (PMP).For absorption, the “phosphorylated” form must be hydrolyzed to “dephosphorylated” form by the enzyme alkaline phosphatase in the intestine.In the portal vein Vit B6 is present as PL, PM, PN.In the liver they are converted back to phosphorylated forms. This conversion is catalyzed by the ATP requiring enzyme, pyridoxal kinase.PLP and PL account for 90% of the total B6 in the blood.In the blood B6 is transported both in the plasma and the RBCs.In the blood PLP is hydrolyzed to PL because only free PL gets inside the cells.In muscle and other tissues, PL is converted back to PLP by a reversible reaction with the help of alkaline phosphatase and pyridoxal kinase.
4 Functions FUNCTIONS: B6 is involved in: Amino acid metabolism Transamination reactions required for the synthesis and catabolism of the amino acids.Decarboxylation reactions.Breakdown of glycogen Glycogenolysis (cofactor for glycogen phosphorylase).80-90% of body vit B6 is present in the muscles, most of it in PLP (coenzyme) form bound to glycogen phosphorylase. Only 1 mol or less is present in the blood,Synthesis of epinephrine (adrenaline) and norepinephrine (noradrenaline)Synthesis of niacin (vitamin B3) from the amino acid tryptophan.
5 Covalent bonds of -amino acids made labile by their binding to PLP-containing enzyme In the reactions of amino acid metabolism, the formyl (CHO) group of PLP condenses with -NH2 group of an amino acid and forms a Schiffs base. This linkage weakens or labilizes all the bounds around the -carbon of the amino acid.The specific bond of an amino acid that is broken depends on the particular enzyme to which PLP is attached.
7 Deficiency Food sources: Requirements: TOXICITIES: In animal foods major forms are PL and and PM along with their phosphorylated forms.In plants PN.Bananas, beans, lentils, walnuts, salmon, chicken, beef, whole grain breads and cereals, soybeans, liver, eggs, dairy products are excellent sources.Requirements:The requirement for vitamin B6 in the diet is proportional to the level of protein consumption ranging from mg/day for a normal adult.During pregnancy and lactation the requirement for vitamin B6 increases approximately 0.6 mg/day.TOXICITIES:Megadoses of B6 (daily doses of >500mg) are used to treat pms symptoms. They can cause neurotoxicity and photosensitivity in some individuals.Deficiencies: are rare and usually are related to an overall deficiency of all the B-complex vitamins.Certain drugs form complexes with PL and PLPPenicillamine (used to treat rheumatoid arthritis and cystinurias).Isoniazid (the hydrazide derivative of isonicotinic acid) is the primary drug for chemotherapy of tuberculosis.
8 7. BIOTINIt is sometimes called vitamin H and also coenzyme R.Biotin is relatively small, bicyclic (two-ring) compound formed from a tetrahydrothiophene (thiophene) ring ,and a second ring, which contains a ureido group.The thiophene ring also has a valeric acid side chain.Although eight different stereoisomers of biotin exist, only one stereoisomer is found naturally and to have biological activity as a coenzyme. It is called d-(+)-biotin, D-biotin or simply biotin.
9 HolocarboxylaseIn humans, the four holocarboxylases are : acetyl-CoA carboxylase, propionyl-CoA carboxylase, pyruvate carboxylase and beta-methylcrotonyl-CoA carboxylase. Biotin is chemically bonded in each of these enzymes via an amide linkage between the carboxyl group of the valeric acid side-chain in biotin and the epsilon-amino group of the lysine residue in the apocarboxylase.The enzyme that catalyzes the formation of this covalent bond is called holocarboxylase synthetase.
10 Biotin CycleBiotin cycle: the chain of chemical reactions involved in the use and reuse of the vitamin biotin. One important role of biotinidase is:1. To separate or free biotin from proteins to which it is bound in foods. Biotin in its free form can then be used by the body.2. Biotinidase lets the body recycle or reuse the biotin over and over again so that we do not need to consume large amounts of this vitamin in our diets.Within cells, the carboxylases (pyruvate carboxylase, acetyl-CoA carboxylase, methycrotonyl-CoA carboxylase, propionyl-CoA carboxylase) are biotinylated via holocarboxylase synthetase. Biotin and apocarboxylases are the substrates. ATP and magnesium also participate in the reaction.Biotinidase deficiency is a treatable, inherited metabolic disorder in which the body cannot process the vitamin biotin in a normal manner.
11 Functions Coenzyme involving CO2 transfer Deficiency acetyl CoA to malonyl CoAPropionyl CoA to methylmalonyl CoAPyruvate to oxaloacetateDeficiencyFatty liver and kidney syndrome, a fatal metabolic disorder in chicks and turkey poultsRaw egg white (avidin) can block absorption
12 Biotin (functions)Coenzyme for several reactions involving CO2 fixation into various compounds e.g.Acetyl CoA to malonyl CoA(acetyl CoA carboxylase)- initial step in de novo fatty acid synthesis.Pyruvate to oxaloacetate (pyruvate carboxylase)Propionyl CoA to methylmalonyl CoA (propionyl CoA carboxylase)
13 Deficiency symptomsRare because of widespread distribution in plant and animal food and significant lower gut synthesis.SourcesYeast, rice, soybeans, peanuts, fish (herring and mackerel), mushrooms and bananas, safflower meal, liver and milk are rich sources.Can be induced by eating raw egg whiteThe fact is that nature created the egg in such a way that its yolk is very rich in biotin. One of the highest concentration in nature. Eat the egg whole together with the egg white and you will be fine.Egg whites contain a glycoprotein called "avidin" which binds biotin - one of the B vitamins - very effectively. The cooking process deactivates the avidin in the egg, much the same way it deactivates every other protein in the egg white.Biotin deficiency is chief cause of fatty liver and kidney syndrome.This baby developed severe biotin deficiency during intravenous feeding without biotin.Aajonus Vonderplanitz,in his book “We Want to live” is a strong proponent of raw eggs.
14 How Biotin Works 1- Biotin carrier protein 2- Biotin carboxylase 3- Transcarboxylase
15 9. FOLIC ACID (folacin)Folacin includes several derivatives of folic acid (monopteroylglutamic acid).Folic acid is obtained primarily from yeasts and leafy vegetables as well as animal liver. Animals cannot synthesize PABA nor attach glutamate residues to pteroic acid, thus, requiring folate intake in the diet.“MicroorganismsOnly cansynthesize Folacin”Sulphonamidesand antibioticsblock PABAincorporationby competitiveinhibition
16 StructureFolic acid exists in a polyglutamate form. Intestinal mucosal cells remove some of the glutamate residues through the action of the lysosomal enzyme, conjugase.
17 Active functional form is: Tetrahydrafolic acid (THF).Humans and microorganisms:Folic acid is reduced within cells (principally in the liver where it is stored) to tetrahydrofolate (THF or H4folate) through the action of folate reductase [or dihydrofolate reductase (DHFR) ] which is an NADPH-requiring enzyme.
19 Active center of tetrahydrofolate (THF) Active center of tetrahydrofolate (THF). The N5 position is the site of attachment of methyl and formimino groups, the N10 the site for attachment of formyl group and that both N5 and N10 bridge the methylene and methenyl groups.
20 Folate conversions Carrier of one-carbon (e.g. methyl) groups The one-carbon units are added to, or removed from, metabolites such as:histidineserinemethioninepurines.
21 FunctionsRole of N5,N10-methylene-THF in dTMP synthesis is the most metabolically significant function for this vitamin.Vitamin B12 and N5-methyl-THF in the conversion of homocysteine to methionine is important in helping cells to regenerate needed THF.
22 Participation of H4folate in dTMP synthesis ______Deoxyuridine______________ ________Deoxythymidine ____Monophosphate (dUMP)_______________Monophosphate (dTMP)_______
23 Deficiency symptoms Identical to those for vitamin B12 deficiency: Deficiency is rare due to the adequate presence of folate in food.Deficiency can occur when there is:Poor dietary habits as those of chronic alcoholics.Impaired absorption or metabolismCertain drugs such as anticonvulsants and oral contraceptives and methotrxate can impair the absorption of folate.An increased demand for the vitamin.PregnancyFolate will nearly double by the third trimester of pregnancy.Identical to those for vitamin B12 deficiency:Effect of folate deficiency on cellular processesis upon DNA synthesis.Impairment in dTMP synthesis and purine synthesisCell cycle arrest in S-phase of rapidly proliferating cells, especially hematopoietic cells.The result is megaloblastic leukemia as for vitamin B12 deficiency.The inability to synthesize DNA during erythrocyte maturation leads to abnormally large erythrocytes termed macrocytic anemia.Deficiency during pregnancy can cause neural tube defects such as spina bifidia.
24 VITAMIN B12 (cobalamin)Vitamin B12, is also called cobalamin, cyanocobalamin and hydroxycobalamin.It is built from :A nucleotide andA complex tetrapyrrol ring structure (corrin ring)A cobalt ion in the center.A R- groupWhen R is cyanide (CN), vitamin B12 takes the form of cyanocobalamin.In hydroxycobalamin, R equals the hydroxyl group (-OH).In the coenzyme forms of vitamin B12,R equals an adenosyl group in adenosylcobalamin.R equals a methyl (-CH3) group in methylcobalamin.Vitamin B12 is synthesized exclusively by microorganisms (bacteria, fungi and algae) and not by animals and is found in the liver of animals bound to protein as methycobalamin or 5'-deoxyadenosylcobalamin.
25 Dorothy Crowfoot Hodgkin Known as the "red" vitamin because it exists as a dark red crystalline compound, Vitamin B12 is unique in that it is the only vitamin to contain cobalt (Co3+) metal ion, which, gives it the red color.The vitamin must be hydrolyzed from protein in order to be active.Intrinsic factor, a protein secreted by parietal cells of the stomach, carries it to the ileum where it is absorbed.It is transported to the liver and other tissues in the blood bound to transcobalamin II.It is stored in the liver attached to transcobalamin I.It is released into the cell as Hydroxocobalamin (see the next slide)In the cytosol it is converted to methylcobalamin.Or it can enter mitochondria and be converted to 5’-deoxyadenosyl cobalamin.Dorothy Crowfoot Hodgkin( )Dr. Stadtman in her lab
27 FunctionsOnly two reactions in the body require vitamin B12 as a cofactor:During the catabolism of fatty acids with an odd number of carbon atoms and the amino acids valine, isoleucine and threonine the resultant propionyl-CoA is converted to succinyl-CoA for oxidation in the TCA cycle.methylmalonyl-CoA mutase, requires vitamin B12 as a cofactor in the conversion of methylmalonyl-CoA to succinyl-CoA.5'-deoxyadenosine derivative of cobalamin is required for this reactionThe second reaction catalyzed by methionine synthaseconverts homocysteine to methionineThis reaction results in the transfer of the methyl group from N5-methyltetrahydrofolate to hydroxycobalamin generating tetrahydrofolate and methylcobalamin during the process of the conversion.
28 Methionine and Folate cycles are interrelated Methionine cycleFolate cycleMethionineSAMMethylacceptorTHFMSmethyltransferasesCH2-THFB12MethylacceptorCH3-THFCH3-HomocysteineSAHCBSB6TransulfurationpathwaycystathionineB6cysteine
29 Deficiency symptomsPernicious anemia in humans (inability to absorb B12 because of lack of gastric intrinsic factor).Neurological disorders due to progressive demyelination of nerve cells.This results from increase in methylmalonyl-CoA.Methylmalonyl-CoA is a competitive inhibitor of malonyl-CoA in fatty acid biosynthesis.Can substitute malonyl-CoA in any fatty acid biosynthesis and create branched-chain fatty acid altering the architecture of normal membrane structure of nerve cells.SourcesSynthesized only by microorganisms, so traces only are present in plants; liver is a rich source.B12 is found in organ and muscle meats, fish, shellfish, dairy products, eggs and in fortified foods like breakfast cereals.
30 Vitamin C (Chemical nature) It is derived from glucose via uronic acid pathway. EnzymeL-gluconolactone oxidase is responsible for conversion of gluconolactone to ascorbic acid.This enzyme is absent in primates, including humans, some bats….The active form is ascorbic acid itself.
31 VITAMIN CVitamin C is L-ascorbic acid, which is a colorless, crystalline acid with strong reducing properties.FunctionsVitamin C has antioxidant properties similar to those of vitamin E,Protects cells from free radicals.Protects iron from oxidative damage, thus enhancing iron (Fe2+) absorption in the gut.The main function is as a reducing agent.It has the potential to reduce cytochrome a and c of the respiratory chain and molecular oxygen and nitrates.It is required for various hydroxylation reactions e.g. proline to hydroxypoline for collagen synthesis (see next slide).
32 Hydroxylation of proline and lysine residues in collagen Vitamin C is required for the maintenance of normal connective tissue as well as for wound healing because synthesis of connective tissue is the first event in wound tissue remodeling.
33 Other activitiesSeveral other metabolic reactions require vitamin C as a cofactor:The catabolism of tyrosine and the synthesis of epinephrine from tyrosine and the synthesis of the bile acids.It is also believed that vitamin C is involved in the process of steroidogenesis.The adrenal cortex contains high levels of vitamin C which are depleted upon adrenocorticotropic hormone (ACTH) stimulation of the gland.
34 Roles in the body Sources Citrus fruits and green leafy vegetables Vitamin C is readily absorbed and so the primary cause of vitamin C deficiency is poor diet and/or an increased requirement.Deficiency symptoms1. ScurvyBleeding gumsSmall red spots on skinRough skinWounds fail to healWeak bones and teethAnemia and infections2. Stress (e.g., infections, smoking)Mechanism unknown, but vitamin C requirements increase during stress3. Common cold?4. Disease prevention?Cancer, heart disease
36 Vitamin B-complex Vitamin Chemical name B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 ThiamineRiboflavinNicotinamide (niacin)Adenine (no longer considered a vitamin)Pantothenic acidPyridoxineBiotinInositolFolacin (folic acid)p-aminobenzoic acid (PABA) / H1L-carnitine / b-hydroxy-g-trimethylammonium butyrate(or choline)Cyanocobalamin
37 8. Inositol (Vitamin Bh) Inositol is part of the vitamin B-complex. Since it is not essential in the human diet, it cannot be considered a vitamin.Inositol is naturally present in foods high in fiber (wheat, legumes, bran, etc).It is required for:For proper formation of cell membranes.For formation of lecithinIt is not a vitamin per se, but it works synergistically with many vitamins and nutrients.functions closely with a B complex vitamins, Choline, folacin, Vitamins B-6 and B-12.Inositol is necessary for proper function of nerves, brain, and muscles in the body.This nutrient is also often used to counter depression; indeed, serotonin needs inositol for proper functioning.Exists in all human cells, where it plays an important role in cell proliferation and differentiation.Low levels of this nutrient may result in depression and some research has shown that increased levels of Inositol appear to be a promising treatment for depression.
38 10. P-amino benzoic acid (Bx) A substance required for the synthesis of folic acid by many organisms.PABA is an essential nutrient for some bacteria and is sometimes called Vitamin BX.Para-aminobenzoic acid is included in the B-vitamin complex, although it is not an essential nutrient for humans and it varies in its activity from other B vitamins.Although humans lack the ability to synthesize folate from PABA, it is sometimes marketed as an essential nutrient under the premise that it can stimulate intestinal bacteria.It also absorbs ultraviolet light and is used as aminobenzoic acid, as a topical sunscreen. Abbreviated PAB or PABA.The potassium salt is used as drug against fibrotic skin disorders.
40 11. L-CARNITINE (B11) L-carnitine Is made in the body from the amino acids lysine and methionine,Is needed to release energy from fat.It transports fatty acids into mitochondria, the powerhouses of cells.In infancy, and in situations of high energy needs, such as pregnancy and breast-feeding, the need for L-carnitine can exceed production by the body.Therefore, L-carnitine is considered a "conditionally essential" nutrient.
41 L-carnitineL-carnitine’s actions appear to be particularly important in the heart.As an example, patients with diabetes and high blood pressure were given 4 grams of L-carnitine per day in a preliminary study.After 45 weeks, irregular heartbeat and abnormal heart functioning decreased significantly compared with nonsupplemented patients.
42 Sources of L-carnitine Dairy and red meat contain the greatest amounts of carnitine. Therefore, people who have a limited intake of meat and dairy products tend to have lower L-carnitine intakes.Carnitine deficiencies are rare, even in strict vegetarians, because the body produces carnitine relatively easily.Rare genetic diseases can cause a carnitine deficiency.Also, deficiencies are occasionally associated with other diseases, such as diabetes and cirrhosis.Most people do not need carnitine supplements. For therapeutic use, typical amounts are 1–3 grams per day.