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PROTEINS M.SC IST SEMESTER ADVANCE NUTRITION-I INTRODUCTION Protein is a complex, high molecular mass organic compound that consist of amino acids. • Proteins were discovered by Jons Jakob Berzelius in 1838. • They are essential for structural and functional use of all living cells. • Proteins also act as an important macro nutrient in the human diet, supplying the body’s needs for amino acids, the building blocks of protein. • CONT….. Amino end COO- Carboxylic end H3 N C H R a carbons Side Chain All amino acids have the same general formula. CHEMICAL COMPOSITION OF PROTEIN All proteins are synthesized from amino acid molecules and 20 different amino acids are used in protein synthesis although a protein can contain many hundreds of amino acid units overall CLASSIFICATION OF AMINO ACIDS Amino acids are the units of proteins. There are 20 amino acids. They are classified into four groups: 9 amino acids are termed as dispensible or essential amino acids and 11 amino acids are non essential amino acids. The amino acids are divided into four groups: 1. Essential 2. Non essential 3. Conditionally essential 4. Limiting amino acids ESSENTIAL AMINO ACIDS: These are not synthesized in the body un adequate quantity. Therefore they have to be supplied through food daily. Inadequate consumption of any one of these essential amino acids may result in negative nitrogen balance. The 9 essential amino acids are: 1. ISOLEUCINE 2. LEUCINE 3. LYSINE 4. METHIONINE 5. PHENYLALANINE 6. THREONINE 7. TRYPTOPHAN 8. VALINE 9. SERINE NON ESSENTIAL AMINO ACIDS: These non essential amino acids are those that can be produced within the human body using carbon skeletons and amino groups from other amino acids. These 11 non essential amino acids are: 1. ALANINE 2. ARGININE 3. ASPARAGINE 4. ASPARTIC ACID 5. CYSTINE 6. GLUTAMIC ACID 7. GLUTAMINE 8. GLYCINE 9. PROLINE 10.TYROSINE 11. HISTIDINE ARGININE AND HISTIDINE are essential for infants as their body is not capable of synthesizing these amino acids. CONDITIONALLY ESSENTIAL AMINO ACIDS: Sometimes a non essential amino acid becomes essential under certain circumstances e.g. The body normally uses the essential amino acid phenylalanine to make tyrosine (a non essential amino acid)but if the diet fails to supply enough phyenylalanine or if the body cannot make the conversion for some reasons, as happens in the inherited disease phenylketouria, then tyrosine becomes conditionally essential. Similar is the case with Cysteine and Methionine. LIMITING AMINO ACIDS: A limiting amino acid is an amino acid that is present in relatively small amounts but below the recommended essential amino acid requirements e.g. wheat has less amount of amino acid lysine. Lysine would therefore be called as limiting amino acids. A vegetarian would therefore have to cope up with lysine requirement by taking a supplement or by eating foods high in lysine, like soybeans and other legumes. NUTRITIONAL CLASSIFICATION OF PROTEINS DEPENDING ON QUALITY S.No. Group 01. Complete proteins: eg egg proteins promote good growth in rats and other animals. Limiting essential amino acids 02. Partially complete proteins: eg. Wheat protein promote moderate growth. Partially lacking in one or more essential amino acids. 03. Incomplete proteins: eg. Gelatin or zein do not promote growth. Completely lacking in one or more essential amino acids. Nil FUNCTIONS OF PROTEINS Growth and maintenance of tissue. Formation of essential body compounds (Insulin, gastrin, growth hormone etc.) Transport of nutrients Regulation of water balance Maintain of appropriate pH. Defense and detoxification (the toxins present in foods are detoxified by enzymes mainly in liver which convert them into harmless substances). Source of energy (though proteins can provide 4Kcal energy per gram like carbohydrates, they are used for energy purpose only when the diet has inadequate carbohydrate and fat. DISEASES DUE TO PROTEIN DEFFICIENCY PROTEIN ENERGY MALNUTRITION Two severe forms of PEM are Kwashiorkor and Marasmus. Kwashiorkor results from an acute deficiency of protein, whereas, marasmus results from a more chronic deficiency. Kwashiorkor:- It is an African word meaning a “Disease of the displaced child” who is deprived of adequate nutrition. Occurring mostly in children between the age of 1 and 3 years, when they are completely weaned (taken off the breast). CONT….. Manifestation and signs of kwashiorkor:1. Odema 2. Moon face 3. Growth failure 4. mental changes 5. hair and skin changes 6. infection 7. Nutrient deficiency 8. Short supply of digestive enzymes, digestive tract lining deteriorates. 9. Blood protein is no longer synthesized, so the child become anemic. 10. Develop fatty liver, caused by lack of the protein carriers that transport fat out of liver. 11. Antibodies to fight off invading bacteria are degraded due to which child becomes more prone to infections. 12. Dysentry 13. Measles - which might make a healthy child sick for a week or two, kills the kwashiorkar child within two or three days. CONT….. MARASMUS:- Marasmus is more common in children below the age of 1 years. Manifestation and signs of Marasmus 1. Severe growth retardation 2. Old man’s or monkey’s face 3. Extreme emaciation 4. loose and hanging skin folds over arms and buttocks 5. Absolute weakness 7. Slow metabolism 8. Muscle wasting including weakened heart muscles 9. Slow brain development, permanent impairing learning ability. 10. There is little or no fat under the skin to insulate against cold Protein deficiency can also cause fatigue, insulin resistance, hair loss, loss of hair pigment, loss of muscle mass, low body temperature, hormonal irregulation, as well as skin elasticity. Severe protein deficiency, encountered only in times of famine, is fatal, due to the lack of material for the body to construct its own proteins. DIGESTION OF PROTEIN Mouth: saliva does not contain any protein digesting enzyme. So digestion of proteins does not occur in the oral cavity. However, saliva can denature the uncooked natural proteins such as that present in raw egg , unboiled milk or uncooked germinating seeds. Digestion of protein in stomach: Action of gastric juice: The gastric glands of the stomach secrete gastric juice. It contains hydrochloric acid, Proenzymes*-Pepsinogen and prorennin. Hydrochloric acid maintains a strongly acidic PH of about 2-3.7 in the stomach.HCL Kills bacteria and other harmful organisms that may be present alongwith food. HCI converts pepsinogen and prorennin into pepsin and rennin respectively. Once pepsin is formed it changes pepsinogen into pepsin. Such an activation is called autocatalytic reaction. Pepsin and rennin are absent in invertebrates. Gastric juice is thoroughly mixed with food until it becomes a semifluid mass called chyme. Various reactions are summarized below: CONT….. HCL 1. Pepsinogen (Proenzyme) pepsin HCL 2. Prorennin (Proenzyme) Rennin Rennin 3. Casein (Milk Protein) Paracasein 4. Paracasein+ Ca Calcium Paracaseinate (Curd) Pepsin 5. Calcium Paracaseinate Pepsin 6. Proteins Peptones Peptones CONT….. Pepsin can digest even collagens of connective tissue fibres, but not keratins of horn, hair, skin or nail. Rennet tablets which contain rennin are extracted from calf gastric mucosa. These tablets are often used commercially for coagulating the casein of milk to curd. It is important to note that adult cows and humans do not secrete rennin. The function of rennin is then taken over by pepsin and other milk- coagulating enzymes. Sometimes chime is squeezed into oesophagus. This causes burning of some cells. It is called heart burn (hyper acidy). FUNCTIONS OF HYDROCHLORIC ACID : It kills harmful bacteria. It provides acidic medium in the stomach for gastric digestion. It changes pepsinogen into pepsin and prorennin into rennin. It softens the food and dissolve the cement materials between the cells of the food in order to make them readily available for enzyme action. It stops the action of salivary enzyme. It is believed to control the opening and closing of pyloric opening of the stomach. DIGESTION OF PROTEINS IN THE SMALL INTESTINE: Action of Pancreatic juice: Pancreatic juice contains proenzymes – trypsinogen, chymotrypsinogen and procarboxypeptidase. All these are concerned with proteins digestion. The bile provides alkaline medium for various reactions. All these reactions are summarized below: CONT….. Enterokinase of Intestinal juice 1. Trypsinogen Trypsin (Proenzyme) Trypsin 2.Chymotrypsinogen chymotrypsin (proenzyme) Trypsin 3. Procarboxypeptidase Carboxypeptidase (Proenzyme) Trypsin 4. Peptones Peptides Chymotrypsin 5.Peptones Peptides Carboxypeptidase 6.Peptides Smaller peptides + Amino acids B. DIGESTION OF NUCLEO PROTEINS : The cellular foodstuffs which are rich in nuclei (liver, kidney, pancrease etc) are likewise rich in nucleic acid and nucleoproteins. HCL in stomach Nucleo protein Removes protein portion which is digested together with other food proteins ribonuclease &deoxyribonuclease Free nucleic acid nucleotides and nucleosides from pancreatic juice in duodenum ph 8.0 Nuclease, nucleotidases, nucleosidases in small peptones, purines and pyrimidines Intestines, ph 7.6 ABSORPTION OF AMINO ACID Absorption in amino acids takes place in small intestine. The process requires energy. The basic amino acids lysine, arginine and histidine share a carrier system with cystine. The dependence of amino acid transport on Na ion suggests a direct interaction between the carrier and Na ion. This is similar to that observed in the absorption of glucose. According to the available information, the amino acid associates with the carrier and Na ion in the microville and the complex travels to the inner side of the membrane where it dissociates, releasing the amino acid and Na ion into the cytoplasm. The carrier returns back and functions repeatedly. The Na ion is then actively transported out of the cell. PROTEIN UTILIZATION 1. 2. Product of protein digestion and tripeptides and amino acid, as well as protein secreted into the gut lumen (enzyme, slouged cells etc.) are available for absorption and are transported in the portal vein to the liver. The liver can deaminate amino acids to carbohydrates or the amino acid can enter the amino acid pool. Homeostasis regulations control the concentration of specific amino acid in the amino acid pool. Body protein synthesis and breakdown is regulated such that in the health, the amount of protein taken in is exactly balanced by protein excreted in feaces, urine and skin. Protein balance studies have demonstrated the zero protein balance in health, the negative nitrogen balance in infection or trauma, and the positive balance in growth and pregnancy. Muscle mass is maintained in steady state with a fraction of muscle protein being destroyed and rebuilt daily using amino acids from the pool. CONT…… 3.Amino acids are also required for synthesis of visceral protein by liver and other tissues. Albumin, transferrin, retinol binding protein, and other serum protein are measured to asses the capacity of the body to synthesize visceral proteins. Albumin and globulin are examples of simple proteins that yield only amino acids on hydrolysis. FACTORS AFFECTING PROTEIN UTILIZATION 1. 2. Amino acid balance: An excess of one amino acid may in certain circumstances, reduce the total nutritive value. It is now recognized that relative amino acid requirements vary with age. Therefore the value of the pattern of amino acids in a specific food or diet towards promoting growth will vary considerably depending on a person’s age. Calorie intake: Protein content of diet cannot be evaluated without consideration of the adequacy of the calorie intake. When the calorie intake drops below a certain critical point, protein will be deaminated and used as a source of energy and cannot be used for the synthesis of tissues. 3.Immobility :The ability to synthesize protein is greatly reduced among people who are immobile. Elderly who are bedridden loose protein mass even when dietary protein and energy seem adequate. These have been a similar problem among astronauts, who lose protein as a result of both weightlessness and immobility in space fight. 4. Injury: An increase in nitrogen loss after injury is well documented. High protein intakes either before or after injury do not prevent this loss. However loses are recovered more rapidly once healing begins. 5. Emotional stability: Emotional stresses such as fear, anxiety or anger increase the secretion of epinephrine from the adrenal gland, which in turn causes a series of changes that result in the loss of nitrogen. SOURCES OF PROTEIN Animal foods are rich source of vital dietary element, while protein is also found in plant foods, such as grain and legumes and in eggs and dairy products such as milk and yogurt. In order to obtain the full range of essential amino acids, one should eat a variety of protein foods. Many people choose meat (beef, pork and lamb) as their main source of protein, and eat it regularly through the week. This is not necessarily the best approach. . CONT….. Animal meats commonly contain excess fat and lack other important vitamins and minerals, such as complex carbohydrates and dietary fiber. Plant foods such as legumes, nuts, seeds and grains also provide protein. Soya products are particularly popular (e.g. tofu) but contain phytoestrogens, which can be harmful in excessive quantities. A combination of plant and animal proteins is recommended for a balanced diet. PROTEIN AS A SOURCE OF ENERGY 1 GM OF PROTEIN ON OXIDATION PRODUCES 4 KCAL OF ENERGY PROTEIN TYPE Enzymes EXAMPLE Amylase Structural protein Keratin, collagen Hormones Insulin, glucagon Contractile proteins Actin, myosin Storage proteins Transport proteins Immunological proteins Ferritin Haemoglobin serum albumin Antibodies Toxins Neurotoxin FUNCTION Promote the breakdown of starch to the simple sugar glucose. Hair, wool, nails, horns, hoofs, tendons, cartilage Regulate use of blood sugar Contracting fibres in muscles Stores iron in spleen Carries oxygen in blood Carries fatty acids in blood Rid the body of foreign protein Cobra venom blocker of nerve functions PROTEIN REQUIREMENTS Protein yields approximately 4 calories per gram, which is the same energy concentration as carbohydrate. The recommended level of protein intake for the general population is 12 to 15 percent of total calories. Therefore, someone consuming 2,000 calories per day has an energy equivalent of 240 to 300 calories (60 to 75 grams) of protein per day. Table: ICMR Recommended dietary Allowances of Proteins GROWTH PROTEIN GRAMS PER DAY Man 60 Woman 50 Pregnant Woman 50+15 Lactation • 0-6 Months • 6-12 months 50+25 50+18 Infancy • 0-6 Months • 6-12 months 2.05/kg 1.65/kg Children • 1-3 yrs • 4-6 yrs • 7-9 yrs 22 30 41 Boys • 10-12 yrs • 13-15 yrs • 16-18 yrs 54 70 78 • 10-12 yrs • 13-15 yrs • 16-18 yrs 57 65 63 Girls FACTORS AFFECTING PROTEIN REQUIREMENTS 1.Exercise Both anaerobic and aerobic exercise affect protein requirements in different ways. Exercise increases the oxidation of amino acids as well as the rate of protein turnover in lean body mass during recovery. 2.Caloric Intake Because protein can be used for tissue repair and synthesis as well as for energy, protein requirements will increase as total energy intake decreases. As total caloric intake is reduced, energy needs may no longer be satisfied by carbohydrate and fat intake alone, necessitating that protein be used to provide energy. 3. Negative Energy Balance For clients pursuing body-fat reduction, body-fat loss goals require that a caloric deficit be maintained until the goal is reached. These individuals seek to modify their body composition. During a negative energy balance, amino acids are used to assist in energy production. This is called gluconeogenesis. The increase in gluconeogenesis is supported by the release of branched-chain and other amino acids from structural proteins to maintain glucose homeostasis during exercise. NITROGEN BALANCE The term nitrogen balance refer to the amount of nitrogen intake in food protein and the amount of nitrogen excretes in the urine. Foreg.1 gm of urinary nitrogen result from 6.25gm of protein. Thus if 1gm of nitrogen is excreted in the urine for 6.25gm of protein is consumed, the body is said to be in the nitrogen balance. The body’s nitrogen balance are indicates how will its tissues are being maintained. There are three state of Nitrogen Balance: 1. Positive Nitrogen Balance 2. 3. Negative Nitrogen Balance Equilibrium CONT…… Positive Nitrogen Balance: If an individual have positive nitrogen balance. It means that person consuming more protein than his body needs. Negative Nitrogen Balance: If an individual have negative nitrogen balance. It means that person’s nitrogen output is greater than his nitrogen input. Equilibrium: If an individual nitrogen has reached a state of equilibrium. It means that person is excreting about the same amount of nitrogen as he is taking in. QUALITY OF PROTEIN Protein quality is a measure of the usefulness of a protein food for various purposes, including the growth, maintenance, repair of tissues, formation of new tissue and production of egg, wool and milk. Protein quality of food: There are two factors which determine the protein quality of food: i) How well the essential amino acid content matches the requirement of the human body. ii) How easily the protein is digested and absorbed into the body for utilization. METHODS FOR DETERMINING THE PROTEIN QUALITY 1.) Biological value : The biological value of protein is defined as the percentage of absorbed nitrogen retained by the body. BV = Nitrogen retained Nitrogen absorbed × 100 For the measurement of BV, the experimental animals, namely weaning albino rats are chosen. They are first fed with a protein – free diet for 10 days. Then they are kept on a 10% protein diet to be tested for BV. Urine and feces are collected for both the periods i.e. protein – free diet and protein diet. Nitrogen is estimated in the diet, feces and urine samples. Biological value can be calculated by the following formula: CONT…. BV = ( N absorbed – N lost in metabolism) × 100 N absorbed BV = [ In – (Fn – Fc)] – ( Un – Uc) × 100 In – ( Fn – Fc) Where In = Nitrogen ingested Fn = Nitrogen in feces ( on protein diet) Fc = Nitrogen in feces ( on protein – free diet) Un = Nitrogen in urine ( on protein diet) Uc = Nitrogen in urine ( on protein – free diet) CONT… This indicator is based on the assumption that more nitrogen is retained when the essential amino acids are present in sufficient amounts to meet the needs for growth. A food with a biological value of 70 or more is considered capable of supporting growth. The higher the biological value the better is the quality of the dietary protein. The biological value provides a reasonably good index for the nutritive value of proteins. But unfortunately this method has an inherent drawback. It cannot take into account the nitrogen that might be lost during the digestion process. For instance, if the ingested nitrogen is 100 mg, absorbed is 10 mg and retained is 8 mg, the BV 8/10 X 100 = 80.This figure is erroneous, since the major part of the protein (90 mg ) did not enter the body at all for utilization. 2.) NET PROTEIN UTILIZATION The biological value makes no allowance for losses of nitrogen in digestion. This is included in the net protein utilization. NPU = Retained Nitrogen × 100 Intake of Nitrogen Miller and Bender (1995) developed a direct method of estimating NPU. Groups of 28 day old albino rats are used. One group is fed on non – proteins diet while other groups are fed on the test diets containing 10% protein for 10 days. The animals are killed at the end of 10 days. Body nitrogen is estimated by Kjeldahl method. CONT… NPU = Body nitrogen of the test group – Body nitrogen of the non – protein group + Nitrogen consumed by by non – Protein group Nitrogen consumed by test group This is equal to BV × availability. NPU can be calculated from nitrogen balance data in man. To measure both quantity and quality of the protein in a diet the net dietary protein value NDpV is used. NDpV = Intake of Nitrogen × 6.25 × NPU op. 3.) NET DIETARY PROTEIN ENERGY RATIO It is often convenient to express the protein content of a food in terms of the percentage of the energy content provided by protein. The protein content of a diet can be similarly expressed and an additional factor given for the quality of the mixed protein. Net dietary Protein energy ratio Protein energy Net dietary intake × NPU op. CONT… The NPU of different protein sources ranges from approximately 40 to 94 with the animal protein sources near the top and that from vegetable protein sources near the lower end. The average Indian diet has NPU of 65. NPU measurements like BV which consider only one intake level and zero, tend to overestimate the nutritional quality of some proteins. The best biological estimated of protein quality are provided by the slope of the intake response line from several points in the range of intakes where the line is linear, it should not include zero protein intake, If carcass N retentions in animals are used in this way, the index is the relative nutritive value and the line of the test protein is related to a standard. 4.) PROTEIN EFFICIENCY RATIO This test consists of feeding weaning ( 21 day old ) albino rats with a 10% test protein diet and recording the gain in body weight for a period of 4 weeks. Protein efficiency ratio is represented by gain in the weight of rats per gram protein ingested. PER = Gain in body weight (g) Protein ingested (g) The PER for egg protein is 4.5; for milk protein 3.0; for rice protein 2.2. PER OF DIFFERENT PROTEINS Diet PER Egg hen Protein intake Gain in body in 4 weeks (g) weight in 4 weeks (g) 28 132 Wheat flour 26 40 1.5 gelatin 20 -4 -0.2 4.7 CONT… This method had been very useful in comparing a new protein source against reference protein such as egg protein and does evaluate other factors such as relative digestibility. The test has some serious limitations. The PER is not a true efficiency ratio because not all the protein is used for growth only that consumed above maintenance. It varies with the food intake. The calorie intake must be adequate. In terms of speed of operation and expenses, however, the PER method is advantageous. 5.) CHEMICAL SCORE This is based on the chemical analysis of the protein for the composition of essential amino acids which is then compared with a reference protein (usually egg protein). The chemical score is defined as the ratio between the quantity of the most limiting essential amino acid in the test protein to the quantity of the same amino acid in the egg protein, expressed as percentage. Chemical score: mg of the limiting amino acid / g test protein mg of the same amino acid / g egg protein × 100 CONT… The chemical score of egg protein, for any one of the essential amino acids, is taken as 100 and the rest of the proteins are compared. Chemical scores of foods are as follows : eggs – 100; meat – 70; milk cow – 65; liver – 66; fish – 60; rice – 60; soybean – 57; Bengal gram – 44; groundnuts – 44; wheat – 42; sesame – 40; and gelatin – 0. NUTRITIVE VALUE OF PROTEINS OF SOME FOOD STUFFS food B.V NPU PER Limiting amino acids Egg 96 96 3.8 Nil Milk 90 85 2.8 SAA Meat 74 76 3.2 SAA Fish 80 74 3.5 Tryp Rice 80 77 1.7 Lys,Thr Wheat 66 61 1.3 Lys, Thr, Maize 50 48 1.0 Lys, Thr, Tryp Bengal gram 74 61 1.1 SAA Groundnut 55 - - Lys, SAA, Thr THANK YOU