CHEMISTRY OF CARBOHYDRATE I- DEFINITION : Carbohydrate are aldehyde or ketone derivatives of polyhydric alcohols or any substances derived from them

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II- Importance of carbohydrates : carbohydrates are widely distributed both in plants and in animal tissues . In plants , they are produced by photosynthesis . Carbohydrates constitute about 60% of our diet . They are important because : 1- They serve as a source of energy e . g . glucose . 2- They form structural elements in animal and plant cell . 3- Carbohydrates may combine with lipids ( glycolipids ) or protein (glucoproteins) , Both enter in the structure of cell membrane and form the ground substances between tissues . III- Classification of carbohydrates : A- Monosaccharides : contain one sugar unit . B- Disaccharides : contain two sugar units . C- Oligosaccarides : contain 3 – 10 sugar units . D- Polysaccarides : contain more than 10 sugar units . IV- Monosaccarides: They are the simplest units of carbohydrate i . e . on hydrolysis , they can not give a simpler form . the general formula is Cn(H2O)n A- Naming of monosaccarides : 1- According to the presence of aldehyde or ketone group : a- Aldoses : monosaccarides containing aldehyde group ( -CHO ) , The suffix –ose means sugar . b- Ketoses : monosaccarides containing ketone group ( -C=O ) ,
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2- According to the number of carbon atoms : a- Trioses : monosaccharides containing 3 carbons . b- Tetroses : 4 carbons . c- Pentoses : 5 carbons . d- Hexoses : 6 carbons . e- Heptoses : 7 carbons . 3- According to both presence of aldehyde or ketone groups and number of carbon atoms : a- Aldotrioses and ketotrioses . b- Aldotetroses and ketotetroses . c- Aldopentoses and ketopentoses . d- Aldohexoses and ketohexoses . 4- System for numbering the carbons : The carbons are numbered starting from aldehyde group as carbon number 1. In case of ketoses the carbon of ketone group group is the carbon number 2.

B- Classification of monsaccharides : 1- Trioses : monosaccharides containing 3 carbon atoms . a- Aldotrioses : Glyceraldehyde “glycerose” . b- Ketotrioses : Dihydroxyacetone . 2- Tetroses : monosaccarides containing 4 carbon atoms : a- Aldotetroses : Erythrose and threose . b- Ketotetrose : Erythrulose . Note : The suffix-ulose means Keto sugar . 3- Pentose : monosaccharides containing 5 carbon atoms . a- Types : 1) Aldopentoses : Ribose , arabinose , xylose and lyxose . 2) Ketopentoses : Ribulose and xylulose .
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b- Importance ( function ) of pentoses : 1) Ribose and deoxyribose enter in the structure of nucleic acids RNA and DNA . 2) Ribose enters in the structure of ATP , GTP and other high energy phosphate compounds . 3) Ribose enters in the structure of coenzymes NAD , NADP and flavoproteins . 4) Ribose phosphate and ribulose phosphate are intermediates in pentose phosphate shunt ( a minor pathway for glucose oxidation ). 5) Arabinose and xylose are constituents of glycoprotein in plants and in animals . 6) Lyxose is a constituent of lyxoflavin isolated from human heart muscle . 7) Xylulose is an intermediate in uronic acid pathway ( a minor pathway for glucose oxidation ) . 4- Hexoses : a- Types : 1) Aldohexoses : glucose , glactose and mannose . 2) Ketohexose : fructose . b- Importance : 1) Glucose is the most importance sugar in carbohydrates : a) Dietary carbohydrates are absorbed in the form of glucose . b) In the liver and other tissues , glucose is converted to all carbohydrates in the body e . g . glycogen , galactose , ribose and fructose . c) Glucose is the major source of energy in mammals . 2) Fructose “ Fruit sugar “ : a) It can be converted into glucose in liver . b) It is the main source of energy in mammals . 3) Galactose : a) It can be converted into glucose in liver . b) Synthesized in mammary gland to make the lactose of milk ( milk sugar ) . 4) Mannose : A constituent of many glucoproteins . 5- Heptoses : As sedoheptulose : is formed in the oxidation of glucose through the pentose phosphate pathway .

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3. 2. here the carbonyl group becomes asymmetric carbon atoms .sugar .Intermolecular reactions occur by subsequent condensation between one of the –OH of aldenol group and the –OH group of C4 or C5 to form ring structure ( hemiacetal structure ).Hydration of aldehyde group to form group ( alcohol ) . g .sugar . glucose which has aldehyde group does not give all the reactions of aldehyde. so it is α . the sugar which has an aldehyde group undergoes the following : 1.If the remaining –OH is on the right side . If the remaining –OH is on the left side . so it is β . In solution . This indicates that the –CHO group must be masked or combined in some way .Ring ( cyclic ) Structure of sugars The simple open formula of sugars fails to explain some reactions e . 4 .

g . g . In Haworth formula. 2) All the –OH groups on the left side in old ring structure are written upwards in Haworth formula . the arrangement of H and –OH groups around carbon atoms is as follows : 1) All the –OH groups on the right side in old ring structure are written downwards in Haworth formula . α and β glucopyranose . B) The 1-4 ring form is called furanose as it is resmbles the organic compound furan e .Pyranose and furanose : A) The 1-5 ring form is called pyranose as it resembles the organic compound pyran e . 5 .4.Haworth and chair forms : Cyclic structure of sugars may be present in the form of Haworth or Chair forms. α and β glucofuranose . 5.

Glucose contains 4 assymmetric carbon atoms .ASYMMETRIC CARBON ATOM : is the carbon which attached to 4 different groups or atoms . e.Optical activity : is the ability of substance to rotate plane polarized light either to the right or to the left . If it rotates it to the left so it is called : levorotatory or “l” or ( . of 99 % of glucopyranose ( 36 % ) are present as α . so it is sometimes named dextrose .) . Any substances containing one or more asymmetric carbon atom shows 2 properties . 3. 1. it is dextrorotatory . A. Fructose contains 3 asymmetric carbon atoms .If the substance rorates plane polarized light to the right so it is called : dextrorotatory or d or ( + ) .Specific rotation : it is the angle of rotation specific for each optically active substance when the concentration of substance when the concentration of substance is 100 g / dl and the length of measuring tube is 10 cm .Glucose in solution is present mainly (99 %) as glucopyranose and (1% ) as glucofuranose. It is levorotatory so it is sometimes called : levulose . optical activity and optical isomerism . V.5º ) and for fructose is ( -91º ) 6 .D form and ( 63 % ) as β – D form . 2.g specific rotation for glucose is ( +52.

for example glyceraldehyde which has one asymmetric carbon and thus 2 optically active forms : L and its mirror image D forms . 7 . 1.Optical isomerism : It is the ability of substance to present in more than one form (isomer) . a sugar may be dextrorotatory ( d ) or levorotatory ( l ) irrespective of its D or L forms . g .Configuration ( Enantiomers ) A)The simplest carbohydrates are monosaccharides trioses . A substance containing one asymmetric carbon atom can exist in a number of isomers = 2n where n is the number of asymmetric carbon atoms . it shows no optical activity ( Provided that the angle of rotation is equal in both sides ) . glucose has 4 asymmetric carbon atoms so the number of its isomers equal 24 = 2 x 2 x 2 x 2 = 16 isomers . However. 1) All other monosaccharides are considered to be derived from reference sugar glyceralddehyde .Resolution : It is the separation of optically inactive racemic mixture into its optically active substance . 4- Reference sugar : It is glyceraldehyde which may be present in (D) form in which –OH group attached to asymmetric carbon atom is on the right side and ( L ) form in which the same –OH group is on the left side . e . g . B.Racemic mixture : It is the mixture containing equal number of molecule of 2 optically active sugars . They are classified into D and L forms according to the position of –OH attached to the carbon atom adjacent to last -CH2OH e . carbon atom number 5 in glucose . Thus . 5. one is dextrorotatory and the other is levorotatory . 2) Most of the monosaccharides occuring in mammal are of D configuration ( form ) .

2. α and β sugars slowly change into an equilibrium mixture which has specific rotation of +52. g . α and β glucose are 2 anomers . 1) α -Glucose freshly dissolved in water has specific rotation of +112 . has specific ratation of +19 . 3) when both anomers are left for some times . B) Anomers : These are isomers obtained from the change of position of hydroxyl attached to the anomeric carbon e .5 8 . Also α and β fructose are 2 anomers .Anomeric carbon and anomers : A) Anomeric carbon : is the asymmetric carbon atom obtained from active carbonyl sugar group : carbon number 1 in aldoses and carbon number 2 in ketoses . 2) β -Glucose when freshly dissolved in water . C) Mutarotation : It is a gradual change of specific rotation of any optically active substance having free aldehyde ( -CHO ) or ketone (C=O) group .

carbons number 2 . One contains keto group (C=O) and the other contains aldehyde group ( -CHO ) . 3) Mannose : epimer of carbon 2 . 2 . Glucose .3. Both are isomers. 2) Galactose : epimer of carbon 4 . 4.Epimeric carbon and epimers : A) Epimeric carbon is the asymmetric carbon atom other than carbon of aldehyde or Ketone gbroup e . 9 . B) Epimers : are isomers resulting from the change of position of groups around the epimeric carbons .Aldose – Ketose isomerism : Fructose has the same molecular formula as glucose but differs in structure formula . 3 and 4 . 1) Glucose has 3 epimeric carbons . glactose and mannose are epimers . g . 3 and 4 of glucose .

glucose gives glucose-6-phosphate and glucose-1-phosphate . g . Reaction of sulphuric acids : This acid is a dehydrating agent . c . 3Reducing sugars : Sugars containing free aldehyde or ketone group can reduce other reagents e .VI. 3All monosaccharides can exist in α and β forms .Physical properties : 1All monosaccharides are soluble in water . 2All monosaccharides show the property of optical activity . glucose gives sorbitol . B. g . galactose gives galacticol. B) These tests are nonspecific .Chemical Properties : 1Oxidation : oxidation of sugars give acids 2Reduction : Reduction of carbonyl group gives the corresponding alcohol e . This compound can condense with αβ -naphthol to B) 10 . 4Reactions with phosphoric and sulphuric acids : A) Reactions of phosphoric acid with monosaccharides gives phosphate esters e . ribose gives ribitol . g . removing 3 molecules of H2O from the sugar giving a compound called furfural .PROPERTIES OF MONOSACCHARIDES : A. they can reduce cupric ions of Fehling and Benedict’s reagents into cuprous ions : Cupric ( blue ) + sugar Cuprous ( red ) + oxidized sugar A) These tests are one of the earlist tests for the presence of sugar in urine of diabetics . Phsphorylated sugars are important intermediates in carbohydrate metabolism . 4All monosaccharides can undergo mutarotation . because they can be reduced also by other hexoses or other reducing compounds as vit .

1Glucose is reduced to sorbitol ( gulcitol ) . B) All D-monosaccharides are fermentable . 3Mannose is reduced to mannitol . glucose is oxidized to gluconic acid .Sugar acids : are produced by oxidation of carbonyl carbon . 11 . glucose is oxidized to glucuronic acid . B. All sugars having free carbonyl group can form osazone crystal . a general test for all carbohyrates . 2Uronic acid : Oxidation of last hydroxyl carbon gives uronic acid e. It is a member of vitamin B complex . This is the idea of Molish’s test . a constituent of vitamin B2 ( riboflavin 6Insitol = cyclitol It is a sugar alcohol derived from glucose . g . 1Aldonic acids : Oxidation of carbonyl carbon to carboxylic group gives aldonic acid e . glucose is oxidized to glucaric acid ( saccharic acid ) . g . last hydroxyl carbon or both . g .Osazone formation : Osazone are characteristic crystals resulting from the reaction of sugars with phenylhydrazine . 5Fermentation : Fermentation is the action of bacterial or Yeast enzymes on carbohydrate .Sugar alcohols : Monosaccharides . 3Aldaric acids : These are dicarboxylic acids produced by oxidation of both carbonyl carbon and last hydroxyl carbon e . 5Ribose is reduced to ribitol . 2Galactose is reduced to galacticol ( dulcitol ) .give a violet ring . 4Fructose is reduced to mannitol and sorbitol . both aldoses and ketoses may be reduced at carbonyl carbon to the corresponding alcohol . SUGAR DERIVATIVES : A. C6H12O6 2 CH3-CH2-OH + 2 CO2 6. A) Fermentation of sugas give ethyl alcohol and CO2 .

1Deoxyribose : occuring in nucleic acid DNA .Deoxysugars : Are sugars in which one of the hydroxyl groups has been replaced by a hydrogen atom i. D. the hydroxyl group is replaced by an amino or an acetylamino group .Amino sugar acids : There are a condensation of amino sugars and some acids .Examples : A) Glucosamine : occurring in heparin and hyaluronic acid .C. 2L-Fucose ( 6-deoxygalactose ) : occurring in glycoproteins . They are occuring in glycoproteins . E.Amino sugars : In these sugars . 2.e one oxygen is missed . Examples include neuramnic acid ( NANA ) and sialic acid which is N-acetyl neuramininc acid . 12 . B) Galactosamine : occurring in chondroitin sulphate .Amino sugars are constituents of glycoproteins . 1. gangliosides and glucosaminoglycans . C) Mannosamine : occurring in neuraminic and sialic acids .

the resulting structure is an O-glycoside and the bond is called acetal link . a galactoside and so on . the resulting compound is glucoside . the resulting structure is N-glycoside . B.VIII.This bond is between the hydroxyl group of anomeric carbon of monosaccharides ( carbon 1 in aldoses or carbon 2 in ketoses ) and another compound which may be : A) Another monosacccharides to form disaccharides glycosides as maltose . C) All sugar-sugar glycosidic bonds are O type linkage .e .GLYCOSIDIC BOND AND GLYCOSIDES : Glycosidic bond : It is hhte bond between a carbohydrate and another compound to form a complex carbohydrate . B) Aglycone i. If the fitst sugar is glucose . 2. noncarbohydrate to form glycoside . if galactose .Examples of glycosides : 1Disaccharides 13 .O and N-glycosides : A) If the monosaccharides is attached to –OH group of another sugar glycogen . lactose and sucrose . B) If the monosaccharides is attached to –NH2 of a glycone . 1.

3Glycolipids : as cerebrosides . B. the linkage is an α-bond . Cardiac glycosides : Aglycone here is steroid 2- II. 5Cellobinose = β-glucose + β -glucose (β 1-4 glucosidic bond ).DISACCHARIDES : These are formed by condensation of 2 molecules of monosaccharides bond together by glycosidic bond . 4Glycoproteins . Maltose is produced during digestion of starch by amylase enzyme 3Properties : Maltose contains free carbonyl ( aldehyde ) group so having the following properties : A) It is a reducing agent ( can reduce Benedict’s reagent) .The most important disaccharides are : 1Maltose = α. C. 3Example : lactose consists of β -glucopyranose and β -galactopyranose . Its general formula is Cn(H2O)n-1 A. If it is in α position . 14 . 2The position of the anomeric carbon of the sugar . 2Isomaltose = α-glucose + α-glucose (α 1-6 glucosidic bond ) . The bond is therefore β 1-4 galactosidic linkage . 6Trehalose = α-glucose + α-glucose (α 1-1 glucosidic bond ) . 3Lactose = β-glucose + β -galactose (β 1-4 glucosidic bond ) . The bond is between carbon 1 of β -galactopyranose and carbon 4 of glucopyranose .Naming of glycosidic bonds : glycosidic bonds sugars are named according to : 1The numbers of the connected carbons . 4Sucrose = α-glucose + β -fructose (α 1-B2 glucosidic bond ) .5- Sugar nucleotide as ATP . GTP and other nucleotides : a glycone here is purines and pyrimidines .glucose + α-glucose (α 1-4 glucosidic bond ) . the linkage is a β -bond .Maltose : Also called malt sugar : 1Structure : It is formed of 2 molecules of α-D glucopyranose linked together by α 1-4 glucosidic bond . If it is in the β position . 2Sources : Malt .

diarrhea and abdominal distension . D. B) It can not be present in α and β forms . so having the following properties : A) It is reducing sugar ( can reduce Benedict’s reagent ) . carbon 1 of α-glucose and carbon 2 of β -fructose are involved in glycosidic bond ) so fructose has the following properties : A) It is not a reducing sugar ( can not reduce Benedict’s reagent ). this leads to its fermentation by intestinal bacteria . 3. E) Lactose is digested by intestinal enzyme called : Lactase into galactose and glucose.Sucrose : 1. C) It can show mutarotation . being formed of 2 molecules of α-D glucopyranose . F.Isomaltose : 1Structure : It is similar to maltose . but linked together by α 1-6 glucosidic bond . 3Properties : The same as maltose . 2Sources : it is the sugar present in milk . its concentration is 7. D) It can form characteristic osazone crystals . 2.Structure : it is formd of 2 molecules of α-D-glucopyranose and β -D-fructofuranose linked by α1 B 2 glycosidic bond .4 g/dl . 3Properties : Lactose contains free carbonyl group . 115 . C) It can be show mutarotation . It may appear in urine in late pregnancy and during lactation . 2Sources : Isomaltose is produced during digestion of starch and glycogen by amylase enzyme . B) It can present in α and β forms . It is also present in pine apple and carrot . In human milk . D) It can not form osazone crystals . D) It can form characterstic osazone crystals .Lactose : Structure : It is formed of 2 molecules of β -Dgalactopyranose and β -D-glucopyranose linked together by β 1-4 glucosidic ( galactosidic ) bond.It can be present in α and β forms . Deficiency of this enzyme stops the digestion of lactose.Properties : sucrose contains no free carbonyl group ( because both the anomeric carbons .Sources : cane and beet sugar . C) It can not show mutarotation . B) E.

Sources : It is obtained by partial hydrolysis of cellulose .glucopyranose linked by β 1----------4 glucosidic bond . it gives a mixture of equal number of glucose and fructose molecules .Cellobiose : 1.Sucrose is dextrorotatory . E) Glucose-Fructose Sucrose Dextrorotatory Glucose + Fructose dextrorotatory levorotatory +52.Structure : It is formed of 2 units of β -D.5 -91 Invert sugar Levorotatory G. 16 . 2. On hydrolysis by inverase ( sucrase ) enzyme . This mixture is called invert sugar and it is levorotatory .

2) Outer layer : called amylopectin . Each chain is composed of 2430 glucose units linked together by α 1-6 glycosidic bonds at the branch points .Homopolysaccharides which contain repeated same sugar units . Sources : It is the most important food source of carbohydrate .Starch ( also called glucoson or glucan ) . A. It constitutes 15-20 % of the granule and formed of non branching helical structure of glucose units linked together by α 1 – 4 glucosidic bonds .POLYSACCARIDES : These are carbohydrates . potatoes . Structure : Starch granule is formed of inner (a) and outer (b) layers : 1) Inner layer : called amylose .Heteropolysaccharides which contain repeated different sugar units . They are classified into : A. it is found in cereals . legumes and other vegetables . 17 . B. formed of more than 10 sugar units . It constitutes 80-85 % of the granule and formed of branched chain .Homopolysaccharides : They include : 1.

Properties : 1Starch gives blue colour with iodine . Amylopectin gives red colour with iodine . 2Partial hydrolysis ( digestion ) by amylase enzyme gives various forms of dextrins . 2- Dextrins : These are hydrolytic products of starch . They are formed of α-glucose units but simpler than starch . They include amylodextrin , erythrodextrin and achrodextrin . 3- Glycogen : ( also called animal starch ) : Structure : It is highly branched chain homopolysaccharide . Each branch is composed of 12-14 glucose units , linked together by 1-4 glycosidic bonds and by 1-6 glycosidic bond at branch point (like amylopectin) . Sources : Glycogen is the storage form of carbohydrates in human and animals . It is synthesized and stored in liver , muscles and tissues . Properties : It gives red colour with iodine . 4- Cellulose : a- Structure : It is long straight non branching chains of glucose units (β -D-glucopyranose ) linked together by β 1---4 glycosidic bond . The chains are strenthened by cross linked hydrogen bonds . b- Sources : Cellulose is the chief constituent of the framework of plant ; leaved vegetables , fruits , wood , cotton , …etc . c- Properties : 1Cellulose gives no colour with iodine . 2Cellulose is insoluble in water . 3Cellulose in diet cannot be digested by many mammals including humans because of the absence of hydrolase enzyme that attachs β -linkage .

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Its presence in diet is important because it cannot be digested , so it will increase the bulk of stool . This stimulates the intestinal movement and prevents constipation . 5Cellulose can be utilized and serve as a source of energy in herbivores because their gut contain bacterial enzymes that can attach β -Linkage . 5- Inulin a- Structure : it is frutosan i.e . formed of repeated units of fructose linked together by β 1-2 bonds . A) Sources : Root of artichokes and other plants . B) Properties : soluble in warm water . C) Medical importance : Inulin clearance is one of a diagnostic tests for investigation of glomerular filtration rate . 6- Chitin : A) Structure : It is a polymer of N-acetylglucosamine linked together by glycosidic bonds . B) Sources : It is important polysaccharides invertebrates . B- Heteropolysaccharides : They include glucosaminoglycans , proteoglycans and glycoproteins . GLYCOSAMINOGLYCANS, (MUCOPOLYSACCHARIDES) : A – Introduction : 1- They are formed of repeating disaccharides units [ acidic sugar amino sugar ]n . The acidic sugar is either D-glucuronic acid or its carbon 5 epimer Linduronic acid . The amino sugar is either D-glucoamine or D=galactosamine in which the amino group is usually acetylated . The amino sugar may also be sulphated at carbon 4 or 6 . 2- Most of GAGs are present extracellulary except heparine . 3- Most of them form the structural components of connective tissue such as bone , elastin and collagen . 4- They act as lubricants and cushion for other tissues because they have the property of holding large quantities of water . 5- When a solution of glycosaminoglycans is compressed , the water is “squeezed out” and the glycosaminoglycans are forced to occupy a smaller volume , when the compression is released , the glycosaminoglycans return back to their original , hydrated volume because of the repulsion of their nagative charges . This property is the cause of resilence of synovial fluid and the vitreous humor of the eye . B- Glycosaminoglycans include : 1- Hyaluronic acid : Structure : Repeated disaccharides unit consists of 1) glucuronic acid .
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2) N-acetylglucosamine . N . B . It is the only GAGs which contains no sulfate group . Site : 1) Synovial fluid . 2) Vitrous body of the eye . 3) Embryonic tissue . 4) Cartilage . 5) Loose connective tissue . C- Function : 1) It permits cell migration during wound repair and morphogenesis , i . e . differentiation of cells in the form of organs and tissues in the early embryo . 2) It makes extracellular matrix loose because of its ability to attract water . 3) It makes cartilage compressible because of its high concentration in this tissue . 4) It acts as a lubricant in joints . d- Role in disease : Hyaluronic acid facilitates cell migration .It is produced in increased amounts by tumor cells . This facilitates migration of these cells through the extracellular matrix and spread of the tumor . 2- Chondritin 4 – and 6 – sulfate : Structure : It is usually present in association with protein to form proteoglycan aggregates . The repeated disaccharides unit consists of : 1) Glucouronic acid . 2) N-acetylgalactosamine with sulfate on either C-4 or C-6 . Site : It is the most abundant GAGs in the body . It is found in : 1) Cartilage , tendons , ligements and bones . 2) Aorta , skin , cornes , umbilical cord and in certain neurons . c- Functions : 1) In cartilage : it binds collagen and hold fibers in strong network . 2) Help to maintain the shape of skeletal system . 3) Have role in compressibility of cartilage in weight bearing . 3- Keratan sulfate : Structure : The repeated disaccharides unit consists of : 1) Galactose ( no uronic acid ) , with sulfate on C-6 . 2) N-Acetylglucosamine with sulfate on C-6 . Site : 1) Cornea . 2) It is found as proteoglycan in cartilage . C- Functions : 1) It plays an important role in corneal transparency .
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keratan sulfate . 2) Selera . Functions : 1) In cornea . contain different sugars and have different shape and size . has the following functions : 1) It is a component of cell membrane and act as receptors . dermatan sulfate . Functions : 1) It act as anticoagulant . 2) N-Acetylgalactosamine with sulfate on C-6 .Dermatan sulfate : 1.Heparine : Structure : The repeated disaccharides unit consists of : 1) Induronic acid with sulfate on C-2 . . skin . it plays together with keratan sulfate an important role in corneal transparency . 2) Glucosamine with sulfate on C-3 and C-6 . hyaluronic acid . 3) It is present in basement membrane of the kidney and plays an important role determining the charge selectiveness of golmerular filtration . Proteoglycans : These are chains of glycosaminoglycans attached to protein molecule e . Sits : 1) Cornea . heart . chondroitin sulfate . g . blood vessels and heart valves .4. They differ from each other in that they present in different sites . 2) Its presence in sclera may play a role in maintaining the overall shape of the eye . They serve as a ground substance and associated with structure elements of tissues as bone elastin and cartilage.Structure : The repeated disaccharide unit consists of : 1) L-Induroic acid . 3) Skin . 21 . 2) Heparine sulfate ( which is the same as heparine in structure except some glucosamines are acetylated and there are fewer sulfate groups ) . 2) It participates in cell adhesion and cell-cell interaction . heparine and heparan sulfate . kidney and spleen .PROTOGLYCANS AND GLUCOPROTEINS : Both are proteins containing carbohydrates . 5. Site : Heparine present in mast cells ( intracellular compound ) Mast cells are located along the wall of blood vessels of liver . The carbohydrate part is present in very long unbranched chains ( more than 50 monosaccharides molecules ) attached to protein core . lungs .

22 . 2) Cell surface recognition receptors : for hormones . 6) They contain no uronic acids or sulfate groups . 4) Plasma proteins : globular proteins – except albumin – present in plasma are glycoproteins . It has free polypeptide portions outside both the external and internal ( cytoplasmic ) surfaces. 5) Most secreted enzymes and proteins ( as hormones ) are glycoproteins.Glycoproteins ( mucoproteins ) : 1. They include : 1) Hexoses : Galactose and mannose . 4) Methylpentose 5) Sialic acid . where they act as protective biologic lubricants .Structure : They consist of : Protein core : Carbohydrate chains which are branched short chain ( from 2-15 monosaccharides units ) such usually called oligosaccharides chain . 3) Glycophorin : Which is glycoprotein present in human red cell membrane . Glycoproteins are components of cell membrane as : 1) Blood group antigens ( A . 2) Acetylhexosamines : N-acetyl glucosamine and N-acetylgalactosamine . B . 2. It spans the lipid membrane . 3) Pentoses : Arabinose and xylose . AB ) . They are components of mucins of gastrointestinal and urogental tracts .Functions : Glycoproteins are components of extracellular matrix . other cells and viruses .

and it does not affect the equilib¬rium constant (i.e. end products) of the reactions.The inorganic catalysts are metals as zinc. so they are denaturated by heat.e.Catalysts: These are organic or inorganic substances that acce¬lerate the rate of chemical reactions. 2. Enzyme structure is not changed by entering the reactions. catalyze one or two reactions only. They accelerate all the biochemical reaction that occur in biologically system which aim to : APrevent uncontrolled and spontaneous reactions .The organic catalysts are enzymes: They are: a. that accelerate the rate of chemical reactions.¬ ENZYME Definitions A. cell.Protein in nature. 1. b. C.e.Highly specific i.Rate of chemical reaction: It is the change in the amount (moles. 23 . They are: a.Substrate: Is the substance upon which the enzyme acts.Enzymes: These are specific protein catalysts. B. b.Non specific i. D. magnesium chloride ions.Not affected by heat. catalyze many reactions. grams) of starting materials (substrates) or products per unit time.

Optical specificity: Enzymes act on one of 2 isomers.Genetic disorders may lead to various diseases.not L. The physical proximity of these various functional groups There are 4 types of specificity a.Genetic expression of enzymes: Enzymes are the major means for the genetic expression 1. pepsin acts on peptide bonds. zyme = yeast). b.Enzyme activity: 24 . They are highly specific.g. The functional group of the enzyme and its co-factor 2. Enzyme specificity: The specificity of the enzyme is determined by 1. B. They act within a moderate PH and temperature range 3.g. urease enzyme acts only on urea.Relative specificity: These are the lowest grade of the specificity in which the enzyme acts on a group of comp¬ounds having the same type of bonds e.Absolute specificity: one enzyme acts only on one subst¬rate e. lipase enzymes act on different triacylglycerols. maltase acts on α-glycosides and not βglycosides. d. Enzymes are biologically active proteins that are synthesized within the General Characteristic of enzymes AProperties of enzymes: 1.BAllow reactions to occur at a rate appropriate to the needs of the cells Substrates -------------------------------.Products ENZYMES: (en = in . The glycolytic enzymes act on D.The set of enzymes in each cell is genetically determined.sugar. . catalyzing only one type of chemical reaction.g.Group specificity: Enzymes need the presence of certain group to act e. c. They are invariably protein 2. 3.The functional group of the substrate 3. C.Living cells contain a unique set of enzymes 2. D.

In such cases. = one carbon group carrier. Lipoic acid.Requirements for enzyme activity: *. *. Some enzymes require metals as Mg2+ and Cu2+as co-factors. FAD and FMN . it is called: holo¬enzyme and its activiy will depend upon: 1) Conformation of the protein which is called a apoenzyme 2) The availability of a non-protein part. They are classified according to their function into 1) Hydrogen carriers: NAD and NADP .¬ Heat liable. *. Stable E. Small molecular weight. which is called cofactor. Big molecular weight. Coenzyme Q. In some enzyme systems. 2) Carriers of groups other than hydrogen: Coenzyme A = acid carrier.1. Differences between apoenzyme and cofactor: Apoenzyme Protein in nature. Cofactor Non protein.Enzymes are either simple or conjugated proteins. Dialyzable.Coenzymes The co-enzymes are heat stable organic compound responsible for the catalytic action of the enzymes.If the enzyme is a conjugated protein. = methyl group carrier. 25 . . Co-enzymes are frequently containing Vit B complex. Some enzymes require coenzymes as NAD+ and FAD.If the enzyme is a simple protein. only the native conformation of the protein is required for activity. the cofactor is tightly bound to the enzyme protein as in case of flavin adenine dinucleotide (FAD). the cofactor is called a prosthetic group. Thiamine diphosphate (TPP) = Acetal group Biotin Pyridoxal phosphate Tetrahydrofolate Cobalamine = CO2 carrier = amino group carrier. Non dialyzable.

Nicotinamide Adenine Dinucleotide (NAD) Nicotiamide Adenine Dinucleotide Phosphate (NADP) The vitamin niacin (Nicotinic acid) is involved in the formation of NAD which acts as electron carrier in oxidation reduction reaction B. Co-enzyme Q or Ubiquinone It is a component of the respiratory chain in the mitochondria and acts as electron carrier 26 .Co-enzymes for transfer of 2 hydrogen A.

Lipoic acid It is involved in the complete system of oxidative decarboxylation of pyruvate or ketoglutarate Co-enzymes for transfer of groups other than hydrogen A. Flavin adenine dinucleotide (FAD) It is the prosthetic group of acyl-CoA dehydrogenase D. Thiamine pyrophosphate (TPP) 27 . Flavin nucleotide co-enzyme Riboflavin (vitamin B2) is the constituent of several enzymes which are involved in intermediary metabolism 1. Riboflavin monophosphate also knowen as flavin mono-nucleotide (FMN): It is the constituent of cytochrome C reductase and the amino acid dehydrogenase 2.C.

It acts as Co-acylase in the acyl transfer reaction. C-Pyridoxal Phosphate Pyridoxine (vit B6) is involved in the enzyme system of transaminases. cystathionine synthetase and heme biosynthesis 28 . Co-enzyme A Pantothenic acid is a component of Co-enzyme A. It is component of the enzyme pyruvate dehydrogenase B.Thiamine (Vit B1) is a component of the co-enzyme TPP that acts as decarboxylase in the oxidative decarboxylation of α-keto acids.

The specific activity is the number of units of enzyme activity per milligram of enzyme protein. Biotin or Vitamin H It acts as co-carboxylase in the transfer of pyruvate to oxalacetate and acetyl CoA into malonyl CoA F. 29 . b.Measures of enzyme activity: a.The unit of enzyme activity: is the amount of enzyme cau¬sing transformation of one micromole (1 µmol) of a subs¬trate per minute at 25 ºC under optimal conditions of measurement.The katal (Kat) is the amount of enzyme activity that transforms 1 mol of substrate per second. c.D.

Lactate .The substrate acted upon.Some enzymes were named by attaching the suffix -ase to the name of the substrates e.Pyruvate + NADH+H E.ENZYMES NOMENCLATURE (NAMING): Enzyme nomenclature has histori¬cally been a source of confusion. 2.(1) : Alcohol e.g.C (1) : Class of enzyme: oxidoreductase. -CHO 3. There are 6 classes of enzymes.The coenzyme involved in the reaction.NAD+ . E. FAD. 1. -OH .C.1.g. B.Fourth digit: indicates the substrate Alcohol Dehydrogenase E. 4. C. maltose & maltase. pepsin.C.1.C.g. (Its old name was lactate dehydrogenase) It catalyze the following reaction Lactate + NAD ----------------------------. 3. 2.g.Trivial names e. trypsin. D.Some enzymes were named according to the type of the reaction e. This number called: Enzyme commi¬ssion numerical code (EC) and it contains 4 digits: 1. This name can indicates: 1.1) : Group upon which the enzyme acts is : CH-OH E.First digit: indicates the class of the enzyme. e.g.Third digit: indicates the coenzyme e. the International Union of Biochemistry (lUB) made a systemic name to each enzyme. the IUB classifies the enzymes by giving each enzyme a number. E.oxidoreductase enzyme.(1) : The coenzyme is NAD+. CLASSIFICATION OF ENZYMES: There are 6 classes of enzymes which are: 30 .In addition to naming enzymes.To standardize enzyme nomenclature. 1.Second digit: indicates the functional group upon which the enzyme acts e. NAD.g.1.The type of reaction catalyzed.g. ethanol is the substrate. 1. aminotransferase. There are several ways of naming enzymes: A.

Example: phosphohexose isomerase: Phosphohexose isomerase_ Glucose-6-phosphate Fructose-6-phosphate 31 . 1Oxidoreductases are further classified according to the sub¬strate oxidized and to the mechanism of oxidation.S (reduced) + Y (oxidized).Dihydroxyacetone Phosphate 5.This group includes: isomerases. Lyases : This group of enzymes catalyzes removal of groups from substrates by mechanism other than hydrolysis. transaminases. Example: Phosphotransferases : Kinases : Glucose + ATP Glucokinase Glucose-6-phosphate + ADP 3. transacylases. transketolases.6 Bisphosphate------------. The mechanism of oxidation is either by addition of oxygen (oxidases) or by removal of hydrogen (dehydrogenases). mutases and epimerases. breakdown of a chemical bond by addition of water: A-B HOH AH + BOH Example: peptidase: 4.AOxidoreductases : This group of enzymes catalyzes an oxidation¬ reduction reaction between two substrates: S (oxidized) + Y (reduced) --------------.g Lactate dehydrogenase * Oxidase: they are group of the enzyme catalyze transfer of hydrogen or electron to oxygen with the formation of hydrogen peroxide e. * They are further classified according to the group transfe¬rred: phosphotransferases. transformylases and transmethylases.Isomerases: This group of enzymes catalyzes the inter conversion of one isomer into the other. * Dehydrogenase are group of the enzymes catalyzes the removal of 2 H or electron from the substrate to the coenzymes e. Hydrolases : This group catalyzes hydrolysis i. Pyruvate Decarboxylase CH3 COCOOH ---------------------------.CH3CHO Fructose 1.g Glucose oxidase * Oxygenase: enzymes catalyzes the incorporation of either molecular oxygen into the substrate e. *.Transferases : This group of enzyme catalyzes transfer of group other than hydrogen SX + Y-----------------------------S + YX .e.g lipoxygenase or one oxygen e.g monoxygenase 2.

2. This is called free energy of the reaction.All the reactions that proceed from initial substrates (ini¬tial state) to products (final state) consume energy. This energy is called activation energy.6.Glutamine + H2O MECHANISM OF ENZYME ACTION: AEnergy of activation: 1. 4.However the substrates do not become products directly.The definition of activation energy: is the amount of energy required to raise all the molecules in one mole of a substa¬nce to the transition state.The effect of enzymes is to decrease the energy of activation of substrates Active site: The substrate is bounded to a specific region on the enzyme called active site which is characterized by Smal portion of the total volume of the enzyme It is composed of R group comes from side chain of the amino acids and the specificity depends on arrangement of these groups 32 . 3. there is a high probability that a che¬mical bond will be made or broken to form the product.At transition state. Example: Glutamine synthetase. . Glutamate + NH3---------------------------. but must be energized (absorb energy) to reach an activated or transition state.Ligases (or synthetases): This group of enzymes catalyzes join¬ing of two substrates using the energy released in the hydroly¬sis of a high energy phosphate compound as ATP or GTP. 5.

2)This is followed by dissociation of this complex into enzyme again and products. E + S------. 1If an enzyme is incubated with its substrate and the appea¬rance of the product with time is recorded on a graph. Catalytic site is presumed to be preshaped to fit with the substrate. there is a temporary combination between the enzyme and its substrate forming enzyme¬ substrate complex. The enzyme changes shape upon binding the substrate. B. The lock and key theory Model of Fischer or template model :The active site of the enzyme is complementary in conformation to the substrate. so that the conformation of subst¬rate and enzyme protein are only complementary after the binding reaction.E + P Two theories have been proposed to explain the specificity of enzyme action a. A.ES ------.Initial velocity: the rate (velocity) at which a reaction pro¬ceeds is measured as the decrease in the concentration of reac¬tants (substrate) or the increase in concentration of the products with time. The induced fit theory Model of Koschland: An essential feature of this model is the flexibility of the catalytic site. This theory postulate that active site of the enzyme has fixed shape.1) During enzyme action. Fig 3 ENZYME KINETICS: It is the study of the rate or velocity of reac¬tions catalyzed by enzymes. the resulting line will have the hyperbolic shape as in the fo¬llowing figure 4 33 . so that enzyme and substrate recognize one another. This occurs at active site of enzyme.

provided that all other conditions remain constant i. the amount of substrate conve¬rted to product is trippled and so on.Thus only in the initial portion of the reaction are the con¬ditions accurately known. Factors affecting enzyme activity: 1Concentration of enzyme: The initial velocity of a reaction is directly proportion to the amount of the enzyme present. which corresponds to the slope of this curve. 3.e.g. The units of velocity are concentration per unit time. For this reason. the amount of substrate converted to product is doubled. Also when the amount of enzyme is trippled. when the amount of enzyme in a reaction is doubled.The decline in the rate of the reaction may be due to deple¬tion of the substrate. e. micromoles per minute. only the initial velocity (Vi) is used in calculating the kinetic parameters of the reaction. is initially constant but gradually decreases. inhibition of the enzyme by its pro¬duct. Each of these events affects the conditions of the reaction. or denaturation of the enzyme. where any further 34 . Fig 5 2. 4.2.Concentration of substrate: The initial velocity of a reac¬tion is directly proportional to the amount of substrate present till it reaches a maximum point known as maximum velocity (Vmax). and in turn affects the velocity of the reaction.The rate (velocity) of the reaction.

At higher substrate concentration. 2) Michaelis and Menten proposed that in any enzymatic reaction.Michaelis-Menten Equation: 1) This equation describes the dependence of reaction ve¬locity on substrate concentration.increase in the amount of substrate causes no increase in the velocity of the reaction. So the rate of reaction will increase. The following diagram can explain: A.E + P 35 . not all enzymes are saturated. the enzyme (E) combines with substrate (S) to form an enzyme-substrate (ES) complex.ES ES then breaks down either to enzyme and substrate again or to enzyme and product (P) K2 ES-------------------. At low substrate concentration. This is true if all other conditions especially enzyme concentration remain constant. K1 E + S -------------------. all enzymes get satura¬ted with substrates and any more increase of substrate concentration will result in no increase in the rate of the reaction (plateau curve).

K-1and K2. Km reflect the affinity of the enzyme for the substrate The smaller Km value. characteristic for every enzyme and paticular substrate.E + S 4) The rates(velocities) at which the partial reactions of this model occur are described by the rate constants K1 . the increase in the initial velocity (Vo) observed with an increase in (S) is due to an increase in the amount of ES formed. K-1 and K2. Km of hexokinase is less than glu¬cokinase 36 . all of the enzyme is involved in an ES complex. [S] and the rate constants K1. Vi = Vmax [S] ------------------------------[S] + Km K1 + K2 -------------------------K-1 Km = If Km = [S] then Vi = Vmax --------------------2 Thus Km can be defined as substrate concentration that produces half maximum concentration Important conclusions about Michaelis-Menten equation Km is a constant. 6) Using the previous model. more active enzyme ( high affinity of the enzyme toward the substrate) Large or high Km value reflect the low affinity of the enzyme toward the substrate Hexokinase is more active than glucokinase because the amount of glucose (substrate) needed to produce ½ Vmax in case of hexokinase is less than in case of glucokinase i. Michaelis and Menten develop¬ed an equation that expresses the initial velocity (Vi) of a reaction in terms of the Vmax . At Vmax . 5) According to this model.K-1 ES-----------------------.e.

3. This increase in reaction velocity is due to the increase in the number of molecules having sufficient energy to pass over the energy barrier and form the products of the reaction.The optimal temperature for enzymatic activity in human body is that temperature similar of that of the cells 37 ºC b.Effect of temperature: a.Lineweaver-Burk plot:Here the reciprocal of V i. the enzyme is inactive. Further elevation in temperature resulted in decrease in reaction velocity. 37 .e 1/V is plotted versus the reciprocal of [S] 1/S. The curve is straight line and it becomes more practical to estimate Km because a few give saturation curve. The re¬action velocity increases with increase of temperature until a maximum velocity is reached.At zero temperature.

*.Here is structural similarity between substrate and inhibitor e.Competitive inhibitors: This type of inhibition occurs when the inhibitor binds reversibly to the active site where the substrate normally would occupy.Dilution of the enzyme-inhibitor complex results in diss¬ociation of the reversibly bound inhibitor and recovery of enzyme activity. pancreatic lipase 7.Reversible inhibition may be competitive or non competitive 2. Effect of pH The optimal pH for an enzyme activity is that pH at which the enzymes acts maximally.Irreversible inhibitors: This type of inhibition occurs when an inhibited enzyme does not regain activity upon dilution of the enzyme inhibitor complex Competitive and noncompetitive inhibitors: 1.these inhibitors bind to the enzyme through noncovalent bond *. The inhibition of the enzymes can be classified either reversible or irreversible 1.Reversible inhibitors:. Increase or decrease in pH.4.5 while pepsin 2 Enzyme-inhibitor Any substance that can diminish the velocity of an enzymatic reaction is called inhibitor. malonic and succinic acids on succinate dehydrogenase 38 . both inhibitor and substrate compete for active site. the ionic state of both enzyme and substrate will be changed and the rate of the reaction decline Each enzyme has its own optimal pH .g. Therefore. *.

Therefore.*. wherease Km is unchanged. the reaction velocity reaches Vmax observed in absence of the inhibitor Effect on Km: A competitive inhibitor increase Km of the substrate i. at a sufficiently high substrate concentration. 39 .e more substrate is needed Effect on Line-weaver-Burk plot: Inhibited and uninhibited reactions show different X axis intercepts indicate increase in Km 2.Effect of competitive inhibitor on Vmax The effect of a competitive inhibitor is reversed by increa¬sing substrate concentration.g heavy metals or oxidizing agent Effect of noncompetitive inhibitors on Vmax : Non competi¬tive inhibition cannot be overcome by increasing the concentration of substrate.Non competitive inhibitors: This type of inhibition occurs when the inhibitor and substrate can bind at different site of the enzyme. Thus. Thus the enzyme show the same Km Effect on Lineweaver-Burke plot: Noncompetitive inhibition is readily differentiated from competitive inhibition by plotting l/V versus l/S and noting that Vmax decreases in of noncompetitive inhibitor. There is no structural similarities between substrate and inhibitor e. Effect on Km: Non competitive inhibitor do not interfere with the binding of the substrate with enzymes. noncompetitive inhibitors decrease the Vmax.

Homotropic enzymes in which the substrate is also the effector b.Heterotropic enzymes : in which the effector is other substance than the substrate Examples of allosteric enzymes Phosphofructokinase is an allosteric enzyme in the glycolytic oxidation of glucose. This results in apparent changes in both Vmax and Km which is reflected in double reciprocal plot Allosteric ( Regulatory) Enzymes 1. It indicates that a molecules called effectors (also called modifiers or modu¬lators) can bind noncovalently at a site other than active site.Effectors may be the end product of a metabolic pathway.Uncompetitive inhibitor This inhibitors binds only on [ES] complex. it is called feed back inhibition. Effectors are positive if they enhance the catalytic reaction and negative if they attenuate the reaction rate. It is allosterically inhibited by high level of ATP and stimulated by ADP Acetyl CoA carboxylase is the allosteric enzyme stimulated by citrate and inhibited by fatty acid Effect of allosteric regulation on Km and Vmax : Allosteric regulation of enzymes causes conformational chan¬ges at the catalytic site. 2.Allosteric enzymes generally catalyze irreversible reaction in metabolic pathways. *-Positive effector : It gives the enzyme a configuration which binds the substrate. If it inhibits the reaction (negative regulation). Classes of allosteric enzymes: there are two major classes a.The term allosteric means "other site". Therefore Michael-Menten does not apply bThey have characteristi sigmoid shape or S-shape curve 40 . This leads to : aAltering Km for a substrate and Vmax . *.

Isoenzymes Isozymes are physically distinct forms of the enzyme with the same catalytic activity: they catalyze the same reaction but at different rates( have different Km value). On the other hand LDH1 (H4) have high Km value low affinity to pyruvate so will prevent accumulation of lactate in heart tissues which function under aerobic condition Serum isoenzymes are valiable in the diagnosis of certain disease serum LDH5 increases in certain liver diseases while LDH1 in heart disease 41 . which is really fit with physiological state of skeletal muscle which mainly function under anaerobic condition.e high affinity for pyruvate so will rapidly oxidize glucose to lactate instead of pyruvate. contains 4 polype¬ptide chains. Lactate dehydrogenase enzyme (LD) which is a tetrameric i.The best known example is: .e. They migrate differently in an electric field. These 4 chains are a mixture of different proportions of 2 chains H and M. For example isoenzyme in heart (H4) and skeletal muscles (M4) have distinctly different kinetics properties which are related to their physiological role in the tissues LDH5 (M4) have low Km i. These two subunit combined in 5 different ways so that there are 5 isoenzymes of LDH Type LDH1 LDH2 LDH3 LDH4 LDH5 Composition H4 H3M H2M2 HM2 M4 Location Heart Heart Brain --Muscles Isoenzymes differ significantly in their Km and Vmax. chains Hand M (H from heart & M from muscle). .

trypsinogen and pepsinogen. Also the enzyme synthesis is inhibited by other substance is called repressor and the process is called repression. Phosphrylation/dephosphorylation : 1. 42 . g. which is stimulated by substance called inducers. Control of enzyme level This is determined by rate of enzyme synthesis. To activate zymogens. Zymogens are inactive because a polypeptide chain masks their catalytic sites. While in case of fasting gluconeogenic enzyme are derepressed by glucocorticoid hormone Proenzymes Zymogens : Some enzymes are synthesized as inactive forms called zymogens or proenzymes e. Induction It is the increase in enzyme production through hormonal activation of the mechanism controlling their gene expression Example: Insulin hormone is hypoglycemic hormone induced the synthesis of group of the enzyme responsible for glucose oxidation ( glucokinase or phosphofructokinase) or glycogen synthesis (glycogen synthetase) Repression and Derepression Insulin hormone is hypoglycemic hormone that represses the bio -synthesis of gluconeogenic enzymes responsible for synthesis of glucose from non carbohydrate sources. The activity of rate limiting enzyme can be regulated in a number of ways 1. and the process is called induction. This can be done by covalent attachment of phosphate group to seryl residue of the enzyme. threonine or tyrosine residues of the enzyme). 2.Regulation of the enzyme activity Control of metabolic regulation of a pathway occurs through modification of the enzymatic activity of the key enzyme in the pathway( rate limiting enzyme).Some enzymes may be regulated by co¬valent modification. most frequently by the addition or removal of phosph¬ate groups from the enzymes (through -OH group of serine. Modulation of catalytic efficiency of the enzyme A-Covalent modification: The activity of some enzyme may be regulated by reversible modification. cleavage of the polypeptide chain should occur to open the catalytic site for its substrate.

De novo synthesis of fatty acids acetyl CoA carboxylase is regulated by feed back nhibition by noncovalent binding of fatty acids It means that the end product of a series of reactions directly inhibits the first enzyme of that series.g ALT and AST in liver disease CPK in heart disease LDH in heart disease 43 .g Blood clotting factors and lipoprotein lipase Non-Functional enzymes They perform no known physiological function in plasma as their names implies. There are two types of plasma enzyme Functional Enzymes They are the enzymes that are normally present in plasma in higher concentration as they perform certain physiological function. Enzyme Compartmentation Compartmentalization of the metabolic pathway allows the separation of the process that proceeds in opposite direction and may otherwise interfere with one another e. It utilizes ATP as a phosphate donor. B. Phosphate groups are cleaved from phosphorylated en¬zymes by the action of phosphoprotein¬ phosphatase. called protein kinase. For example.2. Non covalent modification This is done by non-covalent binding of certain metabolities to allosteric site. Clinical Enzymology Enzyme level determination in plasma can be used ax an index or diagnostic agent in the diagnosis of certain diseases.Depending on the specific enzyme. Their presence in plasma at levels higher than normal level reflect tissues destruction e. phosphorylation of glycogen phosphorylase increases activity. the phosphorylated form may be more or less active than the unphosphorylated enzyme. E. whereas the addition of phosphate to glycogen syn¬thase results in a less active enzyme.Phosphorylation reactions are catalyzed by a family of enzymes.g the anabolic process involved in the biosynthesis of fatty acids from acetyl CoA are located in cytosol while catabolic process concerned with the oxidation of fatty acids are located with the mitochondria. 3.

NAD and NADP.The physiological mediators Cyclic AMP: the second messenger involves in the action of many hormones c.Component of some coenzymes . FAD . b.NUCLEOTIDES The nucleotides are important intracellular molecules having the following functions 1They enter in the structure of the nucleic acids RNA and DNA. 2Purines enter in the structure of a. 3Pyrimidines enter in the structure of 44 .ATP (a source of energy) is the main form chemical energy available. d-S~adenosylmethionine (methyl donner).

hypoxanthine and xanthine. 45 . Polynucleotides----------Nucleotides--------.aUDP-glucose (glycogen and glucouronic synthesis).nitrogen bases. The nitrogen bases are Purine and Pyrimidine acid Purine bases are Adenine .Nucleosides+ PO4 The nucleosides are ---------. cCDP . bUDP-galactose (lactose synthesis). sugar and phosphate.acylglycerol (phospholipid synthesis). guanine .

novo synthesis pathway 46 .SOURCES OF DIFFERENT ATOMS OF PURINE AND PYRIMIDINE BASES Although human ingest dietary nucleic acids and nucleotides. Human can synthesize ample amounts of purine and pyrimidine nucleotides de novo ( from intermediates). survival does not requir their absorption and utilization. Biosynthesis of Purine De.

47 .

*Other tissues are less capable of de novo synthesis of purines. *RBCs leukocytes and the brain are incapable of de novo synthesis of purine.Inhibitors of purine biosynthesis 1Azaserine It is a glutamine antagonist . The conversion of adenine and guanine to AMP and GMP is called PURINE SALVAGE PATHWAY.AMP--------/--------GMP Site of purine biosynthesis: Liver is the major site for purine biosynthesis.Mercaptopurine Blocks the formation of AMP and GMP IMP--------/----------. They depend on exogenous purines (adenine and guanine) for the formation of purine nucleotides (AMP and GMP). 2Diazonorleucine Blocks reaction 2 36. stop purine synthesis at the reaction 5. Regulation of purine biosynthesis 48 .

Mechanism 49 . the exact needs of purines. AMP and ADP i.Purine biosynthesis utilizes 6 ATP molecules and requires glycine glutamine aspartate and methenyltetrahydrofolate.e. GDP. It is allosterically inhibited by GMP. excess purines-inhibit more formation and vice versa. Site Mainly liver. this pathway should be regulated to provide. the end products of purine catabolism is uric acid. To avoid the loss of unnecessary energy and nutrients. The regulation depends upon the activity of PP-ribose-P synthetase enzyme. Catabolism of purines In humans .

Purine over¬production and excretion. At the pH below 5. Regulation of uric acid formation *This depends on the activity of xanthine oxidase enzyme .Sources of uric acid 1.5. *Absence of this enzyme led to decrease uric acid formation. 2-Uric acid produced in the intestine by the action of bacteria on puri¬nes present in diet (1%).Catabolism of purines in the liver (99%). The uric acid is present in the form of a salt sodium urate. Blood . 3-Decrease the rate of uric acid excretion as in nephritis and lead poisoning this leads to renal damage.g. e. CSF. Effect of hyperuricemia 50 . which is present in the liver kidney and intestine.g. CLINICAL DISORDERS HYPERURICEMIA (GOUT) OF PURINE KETABOLISM Hyperuricemia is the increased in blood uric acid level above 7 mg/100 ml. urine. 2-Increase the rate of cell division as in leuckemia ------Purine overproduction and excretion. Causes Hyperuricemia may result either from 1-Increase the activity of PP-ribose-P synthetase-----.75: The predominant form is uric acid e.7 mg/l00 ml The form of uric acid in the plasma and other body fluids depends upon the pH of that fluid At the pH around 7. Plasma level of uric acid 2.

Acute inflammatory reaction called acute gouty arthritis. Here xanthine and hypoxanthine can not be converted to uric acid.formation of crystals of sodium urates--------deposition of these crystals in soft tissues (these urate deposits are called : tophi). 2-Drugs e. PYRIMIDINE BIOSYNTHESIS 51 .acetylsalicylic acid (aspirine) inhibits urate excretion. 2 In kidneys Deposition of uric acid tophi may lead to uric acid stone. HYPOURICEMIA It is the decrease of blood uric acid level below 2 mg/l00 ml. b-Excess xanthine and hypoxanthine excretion (xanthinurea).When the plasma level of sodium urate exceeds certain level (above 7 mg/ 100 ml)-------. 1In Joints Urate crystals will be phagocytosed by leukocytes---------.g. This leads to a-Hypouricemia. The joints that firstly affected are the small joints especially those of big toes. 3-In the cartilage As that of the ear----------. Causes 1-Deficiency of activity of xanthine oxidase enzyme.distruction of cartilage.

*The nucleotides are arranged in chains linked together by a phosphodi¬ester bonds between 5' carbon atom of a deoxyribose of one nucleotide and 3' carbon atom of the next. sugar and phosphoric acid. bInhibited by UTP and other purines. One end contains the phosphate group (5' end) and the other contains the free hydroxyl at 3' end. The nucleotides are linked together by phosphodiester bonds between 3' –hydroxyl on the sugar of one nucleotides and the 5'-phosphate on the sugar of the another nucleotides. Phosphodiester bond = one phosphate is linked to 2 sugars----2 ester bonds. Structure *DNA is formed of many nucleotides (it may reach millions of nucleotide. DNA :. NUCLEIC ACID Nucleic acid are polymers of nucleotides. Each nucleotide is formed of base . guanine . The bases are : Adenine . In long chains formed. The sugar is deoxy-ribose. units).Regulation of pyrimidine biosynthesis Carbamoyl phosphate synthetase II is aStimutated by PP-ribose-phosphate. Deoxyribonucleic acid Site : Nucleus. cytosine and thymine. The chain begins with 5' and ends with 3' 52 .

4 A and there is a complete turn of the helix every 10 base pair. So the bases are not part of the backbone but lie between the 2 strands. The helix is 20µ in diameter. the bases are separated by 3. 53 . The sugar and phosphate molecules form the backbone outside while the bases are arranged inside. In crystalline DNA.*DNA is formed of two antiparallel double chains (strands) of nucleotides in the form of double helix.

Sense and anti-sense strands The 2 strands of DNA molecules are anti-parallel. Pairing rule It was found that in DNA molecules the concentration of deoxyadenosine nucleotides (adenine-ribose~P) = Deoxy-Thymidine nucleotides (thymine-ribose -P) . Also non histone proteins and possibly RNA affect the packing of DNA. Histone are divided into five classes H1 H2A H2B H3 and H4. Histones are small proteins rich in positive charge amino acids (Basic amino acids as arginine and lysine). the opposite strand might be considered comple -mentary or anti-sense strand. one strand runs in the 5' to 3' direction and the other strand runs in the 3' to 5' direc¬tion. Accordingly . i. one strand is considered as the sense strand .e. The pairing of bases of both strands of DNA are complementary and not identical always adenine is paired only with thymine and Guanine is paired only with cytosine A------T G------C Shape of DNA: Chromosomal DNA may be linear or circular DNA is complexed with histones in eukaryotic chromosome which profoundly influence the structure of the chromosome. When single DNA is complexed with histones it forms chromatin threat.The genetic information of DNA is stored as a sequence of purine and pyrimidine nucleotides in the DNA molecule. This resembles 2 parallel streets each going one way but in opposite directions. 54 . and the concentration of deoxy-guanosine nucleotides (guaninerihose-p) =Deoxy-cytidine nucleotides (cytosine-ribose-P).

replication of DNA allows the daughter cells to have the same DNA formation of the mother cells. Then each one acts as a template upon which a new strand is formed. Steps of replication 1.TTP and CTP). using ATP hydrolysis as a source of energy 55 . 2) Transcription : DNA is the source of information for the synthesis of all protein molecules formed in the same cell. Replication is semiconservative process i. By using free nucleotides which are present in the nucleus (ATP GTP . The parent strands unwind at a unique site (called oriC site) with the help of an unwinding helicase enzyme. This can be done by a process called transcription. the more difficult is to separate the 2 strands Functions of DNA 1) Replication of DNA (=Reproduction) The genetic information are stored in the nucleotide sequence of DNA of parent cell passes to daughter cell.Bonds between the bases In double stranded DNA1 adenine is paired with thymine by 2 hydrogen bonds and guanine is paired with cytosine by 3 hydrogen bonds. The higher the G-C content of a DNA molecule . each nucleotide that incorporated in the DNA molecule utilize 2 high energy phosphate. REPLICATION OF DNA It is the process for transferring genetic information on DNA from generation to generation. there is always one strand of old DNA and the other is newly synthesized. Semiconservative replication During cell division the double strands of DNA molecule are separated. So the new DNA is typically identical to that of parent cell. The 2 strands of parent DNA are separated from each other. each strand of DNA induces the formation of a complementary stra¬nd which is identical in structure to the one which has separated This reactions need a large amount of energy i. Thus GC bond is stronger than A -T bond.e.e on the two strands of DNA molecules. When a cell divides to give 2 daughter cells.

The leading strand is synthesized continously by DNA polymerase III that can bind nucleotide only from 5' ---3' direction.The exosed bases on the separated strands serve as a templete for the new strand 4. The enzyme replicate the other strand by turning it on its back (3' to 5' direction Turned on 5' to 3' direction). Because there is no DNA polymerase III that can bind nucleotide from 3' ---5' direction.2. 6. Another protein. a single strand binding protein binds to the unwind DNA to prevent reannealing 3. The replication is continous in one strand “leading strand” and discontinous for the other “ lagging stand” 5.DNA polymerase I fills the gap between the fragments by nucleotides then DNA ligase cause ligation of the fragments to form continous strand 56 . It is called Okasaki fragments 7.

Messenger RNA mRNA . RNA is differ from DNA in the following features RNA DNA Sugar Ribose Deoxyribose Base Uracil Thymine Strands Single Double Stability Breaks down at high pH Resistant Due to presence of 2 OH Messanger RNA mRNA is a one stranded nucleic acid. It encodes the information directing protein synthesis and make up about 21% of RNA in the cell. It is formed under the control of DNA in the nucleus ( transcription ). guanine .4000 nucleotides. Formation of mRNA from coding (sense) strand of the DNA is the first step in the transcription It is composed of 400 . 57 .Ribonucleic acid = RNA There are 3 types of RNA .Transfer RNA tRNA -Ribosomal RNA rRNA All are formed in the nucleus under the control of DNA and the enzyme RNA polymerase. Phosphate Function mRNA is a nucleic acid which carries the genetic information from DNA of the nucleus to the ribosomes where the proteins are synthesized. Structure Bases Adenine . cytosine and uracil Sugar Ribose.

The tRNA is formed as large precursors which are acted upon by ribonuclease P to reduce its size. The ribonuclease P removes 18 ribonucleotides from tRNA precursor close to the anticodon region. Sugar : Ribose.Transfer RNA = tRNA: * It is a single strand of nucleotides formed of Bases Adenine guanine cytosine and uracil. tRNA is formed in the 58 . It has the shape of clove leaf Transfer RNA (tRNA) serves: as adapter molecule for translation of mRNA into amino acid sequence in protein synthesis. Phosphate * This single strand is arranged to form 3 loops and 2 free ends as shown * It is the amino acids carrying RNA.

RIBOSOMAL RNA = (rRNA ) *The cytoplasmic ribosomes are the site of protein synthesis i.nucleus and pass to the cyto¬plasm. So for each codon in mRNA specific tRNA will be attached which means a specific amino acids. tRNA has 2 different regions the anti¬codon region and the terminal region for amino acid which has three nucleotides at 3 terminus CCA.e. *rRNA are formed in the nucleus as a large precursor molecule. this precursor will undergo alteration to be converted to ribosome subunits. There are at least 20 species of tRNA in every cells at lease one to each of the 20 amino acids . Amino acids attaches to 3-OH of the terminal adenine -The middle arm contain the anticodon region which is formed of 3 bases complementary to a cer¬tain codon on mRNA. This rate is measured by what is called Svedberg units Whole ribosome Large subunit Small subunit 59 .So tRNA are the carriers of amino acid. *Each ribosome consists of 2 subunits : one about twice the size of the other. *The whole ribosome and each subunit have its own sedimentation rate. *Ribosomes contain only proteins and RNA (rRNA). In the nucleolus . They contain enzymes needed for this process.

PROTEIN BIOSYNTHESIS Principle of protein biosynthesis * Body proteins are of various types plasma proteins contractile proteins of muscle all enzymes . while the small subunit (40 S) is the binding site for mRNA.Eukaryotes S Prokaryota 30 S 80 S 70 S 60 S 40 50 S The large subunit (60 S) is the binding site for t RNA . some hormones * Protein biosynthesis is subdivided among the various tissues. Codons and Genetic Code Codon is the sequence of 3 nucleotides (bases) in DNA molecules which determines the type and position of the amino acid that enter in the structure of protein molecules Twenty different amino acids are required for protein synthesis. while RNA is formed under the control of DNA. * DNA are present in chromosomes . Mamary gland makes milk proteins proteins Liver makes plasma proteins gamma globulin Muscle makes contractile Lymphocytes make * Nucleic acids (DNA andRNA) are molecules concerned with protein biosyn thesis. The function of this protein depends on the sequence of these amino acids. So there must be at least 20 different genetic codes for the amino acids. DNA or mRNA contains only 4 different 60 . Proteins are composed of amino acids arranged in certain sequence. These information are represented by the sequence of bases in DNA and translated to sequence of amino acids in the cytoplasm. Genetic informations about protein biosynthesis are present in the genes of DNA molecule in the nucleus.

For each amino acid there is specific activating enzyme called : the aminoacyl-tRNA synthetase e.g. 3Translation. STEPS OF PROTEIN BIOSYNTHESIS This occurs in 3 steps 1Activation of amino acids and their carriage by tRNA. alanine 61 . ACTIVATION OF AMINO ACIDS This process occurs in cytosol. do not code for any amino acids. Two codons ( Met+ Init) and (Valine + Int) are used for initiation as well as for methionine and valine In this way the genetic information are sent from the nucleus to cyto¬plasm where the process of protein biosynthesis occurs. 2Transcription of mRNA.nucleotides. If the genetic code is represented by two nucleotides only. we can have 4x4 =16 code words but the genetic code is represented by 3 nucleotides and can provide x3 4x4x4 = 64 3 codons which are called non-sense codons. This exess in the genetic code is explained by the fact that many amino acids are represented by more than one genetic code. Three codons (term*) indicate the termination of polypeptide chain. This leaves 61 codons for 20 amino acids i.e the number of codons is about three times the number of amino acids.

DNA is used only as a templet and unchanged after transcription Transcription begins by recognition of the RNA polymerase -enzyme to the initiation point of the genome. They help the trans¬fer of different amino acids to tRNA as follows -The amino acid becomes attached to the terminal nucleotide of the tRNA (the base of which is adenine).acyl-¬tRNA synthetase. glycine acyl-tRNA synthetase. Transcription Transcription is the synthesis of mRNA of a certain protein under the control of RNA polymerase Transcription is local. 62 . Transcription is conservative. This initiation point is called promoter. This area of DNA is a genome or structural gene. It means that it occurs in a certain area of DNA that carries the genetic infor¬mation (=codons) about this protein(s). The aminoacyl-tRNA is travelled to the ribosomes where protein biosyn¬thesis occurs.

This occurs anti-parrallel to the sense strand that acts as a tempelate for it. The other DNA strand is termed : antisense strand.one strand acts as a tempelate upon which mRNA is formed. 63 . one strand of DNA molecule will act as a sense strand for one gene and antisense for another gene. This strand is termed: sense strand.This is the signal after which the 2 strands of DNA molecule in the genome are separated from each other. Then under the control of RNA polymerase enzyme . Moreover . The formation and elongation of RNA molecules begins by its 5' end and elongates towards its 3' end.

2σ is responsible for recognition and binding of RNA polymerase to the initiation site while the core 2α and 2β that are very rich in GC nucleotides are responsible for elongation of RNA. ATP--------------------------------AMP RNA molecule ends at a certain site in the sense strand. e. RNA polymerase holoenzyme is formed of 5 subunits 2α. The newly formed RNA leaves the nucleus to the cytoplasm (ribosomes) where protein synthesis occurs.The high energy phosphate compound. e. 1Binding of RNA polymerase enzyme to the promoter sequence site. This consumes a large amounts of energy.g. Summary of transcription 1. 7The newly formed RNA leaves the nucleus to the cytoplasm (ribosomes) where protein synthesis occurs. 2β and 2σ. CTP and U'TP act as a donners for different nucleotides needed for the formation and elongation of RNA. 3At the sense strand of the genome . GTP. CTP and UTP act as a donner for different nucleotides needed for the formation and elongation of RNA. ATP. Posttranscriptional changes of RNA molecules 64 . This site is termed stop site. the enzyme select the correct ribonucleotides and catalyze the formation of RNA molecule which begins at its 5' end.AMP 6RNA molecule ends at a certain site in the sense strand. 4Then elongation of the RNA molecule from the 5' to the 3' end continuous antiparrallel to its tempelate. This consumes a large amounts of energy. ATP---------------. 2The 2 strands of genome in the DNA are separated into sense and anti-sense strands (enzyme unwinds a short stretch of DNA). 5The high energy phosphate compounds ATP .g. This site is termed : stop site. GTP .

which is cleaved to give 2 fragments (40S) and (60S) so.INITIATION * mRNA binds to 40S ribosomal subunit. This addition helps the transfer of mRNA from the nucleus to the cytoplasm. 65 .g. Many of these ribosomes can aggregate to translate a single mRNA . TRANSLATION (Protein synthesis) The mRNA migrates from the nucleus to the cytoplasm (ribosomes) where protein biosynthesis occurs. and is called polyribosomes. changes of RNA after their synthesis mRNA together with tRNA and rRNA undergo some modification after their transcription and before they are released from the nucleus to the cyto¬plasm. 2.i. r RNA is synthesized as one simple long strand . * Translation of genetic code that are carried by mRNA to the ribosome can be divided into 3 stages aInitiation b-Elongation cTermination a..Cleavage of the transcripted RNA e.e. This binding needs a protein fac¬tor which is termed : initiation factor 3 (IF-3). * The ribosome is the cellular structure on which various mechanisms occur to assemble the protein molecule. These changes may be 1Addition of some new nucleotides (AMP) to the 3' end of the transcripted mRNA. the rRNA molecules are ready to be incorporated into the ribosome.

3 and hydrolysis of GTP into GDP +Pi. The formation of 80S ribosome is thus complete. 2 .complex This complex together with a third protein factor which is termed initia¬tion factor 3 (IF-l) will attach the anticodon of t-RNA to the first codon (on mRNA) to form an initiation complex with 40 S ribosomal subunit. The initiation complex attaches with 60 S ribosomal~subunit with release of initiation factors 1 . 66 .*The aminoacyl-tRNA interacts with GTP and another protein factor that is termed : Initiation factor 2. (IF-2) ----.

Complex Aminoacyl tRNA site A The free NH2 group of amino acid 2 (aa2) attracts the COOH group of the amino acid 1 (aa1). b. 67 . *The first amino acid in the polypeptide chain (first aminoacyl tRNA)gets attached to the P site. Aminoacyl tRNA enters A site as follows Aminoacyl tRNA + Elongation factor 1 (EF-1) + GTP--------. These are P site (Peptidyl site) and A site (aminoacyl site). GTP and elongation factor 2 . the ribosome moves in the direction of a 5'------3' direction on mRNA to translocate the newly formed peptidyl tRNA in site P.ELONGATION The binding of the proper aminoacyl tRNA in A site depends on proper codon recognition. In the presence of the translocase enzyme . The subsequent aminoacyl tRNA are attached to A site. Site P is now free. This leads to the transfer of peptide chain to site A under the influence of peptidyl transferase enzyme with release of the peptidyl tRNA.*The complete ribosome contains 2 sites for tRNA molecules.

Immunoglobulin.g.g. REGULATION OF PROTEIN SYNTHESIS : OPERON MODEL Operon can regulate protein biosynthesis .TERMINATION After many cycles of elongation . POST-TRANSLATIONAL REGULATION Some proteins after being translated undergo additional changes to be active. Operon is a group of compo¬nents in DNA that are essential for the transcription of structural 68 . It occupies by a third aminoacyl~tRNA according to codon-anticodon and the process is repeated. Releasing factor hydrolyzes the bond between the peptide and tRNA at site P . the terminating codon of mRNA is reached at site A. Conversion of proline to hydroxyproline. There is no tRNA with anticodon to recognize the terminating codon of mRNA.* Site A is now free. The polypeptide chain is thus increased each time by one amino acid. These changes may be 1Addition of carbohydrate To form glycoprotein e. C. and release the protein molecule. 2e. 3Conversion of inactive to active form proinsulin is converted to active insulin by removal of C chain.

2Operator. This repressor has a high affinity for the operator. 69 .Opera¬tor is free ------------. The operator is between the promoter site (at which the RNA polymerase is attached) and the structural genes (G1 . this prevents the transcription of the structural genes. Inducer is a substance that can bind with the repressor -------. These components are 1Promoter. Each component is formed of a number of nucleotide in DNA molecule. G2 G3).Promotor together with RNA-polymerase enzyme will initiate transcription of structural genes to synthesize proteins. Examples of inducers and repressors 1Clucocorticoids act as inducers for the synthesis of gluconeogenic enzv¬mes. 3Structural gene(s). When repressor is atta¬ched to the operator . Structural genes Regulator gene Promotor Operator Transcription of regulator gene leads to the formation of a protein termed : repressor. 2Insulin acts as an inducer for glycolytic enzymes and as repressor for gluconeogenic enzymes. 4Regulator gene.gene(s) of certain protein(s).

70 .

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