Extraction of Amylase Enzyme From Yam | Experiment

Extraction of Amylase Enzyme From Yam | Experiment Amylolytic enzymes are widely distributed in plant tissues, e.g. in storage tissues such as seeds and tubers and in vegetative organs such as leaves. There exist two types of amylases in some species of plants, (E.C.3.2.1.1; 1-4-ÃŽÂ ±-D-glucan glucohydrolase) and (E.C.3.2.1.2; 1-4-ÃŽÂ ²-D-glucan maltohydrolase) amylases [Thoma, J.A., J.E. Sprandlin and S. Dygert, 1971]. Beta-amylase (-1, 4-glucan maltohyrolase, E.C.3,2,1,2) is an exoamylase that attacks the non reducing ends of starches molecules, producing a-maltose and a limit dextrin as products [Thoma, J.A., J.E. Sprandlin and S. Dygert, 1971]. In starch-enriched tissues, ÃŽÂ ²-amylase may play a role in the mobilization of starch during germination or sprouting tubers [Greenwood, C.T. and E.A. Milne, 1968]. Many reports have been demonstrated that ÃŽÂ ²-amylase has a great commercial value in food and beverage industries. The enzyme is useful in structural studies of starch and glycogen. Marshal and Whelan [Marshall, J. et al 1973] report on the removal of any contaminating ÃŽÂ ²-glucosidase. The practical interest of ÃŽÂ ²-amylase was concentrated on its capacity to produce maltose syrups from starch [Biovin, P., 1997.]. ÃŽÂ ²-amylase has previously been purified and characterized from different types of plant sources and a few of microbial origin. In higher plants, the molecular characterization of ÃŽÂ ²-amylase has been carried out on enzyme purified from the organs enriched in starch such as sweet potato tubers [Balls, A.K.,1948, et al], leaves [Vikso-Nelson, A., et al 1997], bulbs [Dicko, M.H., et al, 2000], seeds of various cereal species such as barley [Shinke, R. et al 1971], wheat [Trachuk, R. et al 1966], rice [Okamato, K. and T. Akazawa, 1978] and other higher plants such as soybean [Gertler, A. and Y. Birk, 1965]. On the other hand, much less information is available on the purification and characterization of ÃŽÂ ²-amylase from root. The present study reports the pur ification of ÃŽÂ ²-amylase from Yam (Dioscorea esculenta) root to a pure state along with its characterization. Amylase is an enzyme that breaks starch down into sugar. Amylase is present in human saliva, where it begins the chemical process of digestion. Foods that contain much starch but little sugar, such as rice and potato, taste slightly sweet as they are chewed because amylase turns some of their starch into sugar in the mouth. The pancreas also makes amylase (alpha amylase) to hydrolyse dietary starch into disaccharides and trisaccharides which are converted by other enzymes to glucose to supply the body with energy. Plants and some bacteria also produce amylase. As diastase, amylase was the first enzyme to be discovered and isolated (by Anselme Payen in 1833). Specific amylase proteins are designated by different Greek letters. All amylases are glycoside drolases and act on ÃŽÂ ±-1,4-glycosidic bonds. It will start to denature at around 60C. Amylase digests not only carbohydrates but also dead white blood cells. For example, when you are low in amylase you are a candidate for abscesses (inflamed areas with pus but not bacteria). If you have a toothache and are being treated with antibiotics, but it doesnt go away, chances are you have an abscess. Amylase is involved in anti-inflammatory reactions such as those caused by the release of histamine and similar substances. The inflammatory response usually occurs in organs which are in contact with the outside world, i.e., the lungs and skin. These include skin problems such as psoriasis, eczema, hives and all types of herpes. Some lung problem including asthma and emphysema may require amylase plus other enzyme formulas depending on the particular ailment. There are many types of amylases, but of importance are: ÃŽÂ ±-amylase, ÃŽÂ ²-amylase and glucoamylase. A diagram of an amylase molecule from human saliva. 1.1 ÃŽÂ ²Ãƒâ€šÃ‚ ­ Amylase (EC 3.2.1.2) (alternate names: 1,4-ÃŽÂ ±-D-glucan maltohydrolase; glycogenase; saccharogen amylase) Another form of amylase, ÃŽÂ ²-amylase is also synthesized by bacteria, fungi, and plants. Working from the non-reducing end, ÃŽÂ ²-amylase catalyzes the hydrolysis of the second ÃŽÂ ±-1,4 glycosidic bond, cleaving off two glucose units (maltose) at a time. During the ripening of fruit, ÃŽÂ ²-amylase breaks starch into maltose, resulting in the sweet flavor of ripe fruit. Both ÃŽÂ ±-amylase and ÃŽÂ ²-amylase are present in seeds; ÃŽÂ ²-amylase is present prior to germination, whereas ÃŽÂ ±-amylase and proteases appear once germination has begun. Cereal grain amylase is key to the production of malt. Many microbes also produce amylase to degrade extracellular starches. Animal tissues do not contain ÃŽÂ ²-amylase, although it may be present in microrganisms contained within the digestive tract. 1.1.2 CARBOHYDRATE METABOLISM Digestion of carbohydrate begins in the mouth by the action of salivary ÃŽÂ ±-amylase.Only limited digestion of carbohydrate occurs, however, because salivary ÃŽÂ ±-amylase is denatured in the stomach due to the low pH. Digestion begins again in the small intestine when pancreatic ÃŽÂ ±-amylase is secreted. Starch is broken down into maltose, isomaltose, and maltotriose by ÃŽÂ ±-amylase through the hydrolysis of ÃŽÂ ±-1-4 glycosidic bonds. These products as well as any other disaccharides that were ingested must be further digested to their respective monosaccharide units by brush border enzymes (maltase, isomaltase, lactase, and sucrase) before absorption. Maltose is hydrolyzed to two glucose molecules by maltase. Isomaltose is hydrolyzed to two glucose molecules by isomaltase. Lactose is hydrolyzed to one molecule of glucose and one molecule of galactose by lactase. Sucrose is hydrolyzed to one molecule of fructose and one molecule of glucose by sucrase. After absorp tion, glucose, galactose, and fructose are transported to the liver via the portal blood. The liver can transform galactose and fructose into glucose (Gropper et al 2005). 1.1.3 REACTIONS OF BETA AMYLASE Starch + H2O in vitro breakdown of semicrystalline starch particles by beta-amylases increases significantly if they act together with glucan, water dikinase starch substrate of different sources, e.g. wheat, wheat bran, rice bran beta-amylase hydrolyzes alpha-1,4-linkage, raw starch granules from potato, wheat, rice and corn, with the granules from rice being the best substrate, beta-amylase attacks very slowly on the starch granules, hydrolyzes corn granules efficiently at 45 °C . Beta-amylase is an exo-enzyme that catalyzes the hydrolysis of the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, Glu-172 and Glu-367 are catalytic residues, substrate recognition mechanism, enzyme structure beta-amylase is an inverting enzyme that hydrolyzes the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, catalytic mechanism, Glu-172 acts as general acid, Glu-367 acts as general base catalyzes the hydrolysis of alpha-1,4-glucosidic linkages of soluble starch, and liberates beta-anomeric maltose from the nonreducing ends, exo-acting enzyme, composed of two functional domains, a catalytic domain: domains A and B, and starch-binding domain: domain C, beta-amylase has three carbohydrate-binding sites aside from the active site: two in domain B named Site2 and Site3, one in domain C named Site1, roles of these sites in the catalytic reaction and raw starch-binding, beta-amylase hardly h ydrolyzes raw starch from wheat, corn, potato or sweet potato, but binds to it strongly hydrolyzes the alpha-1,4-glucosidic linkage liberating beta-maltose from the non-reducing end of substrate, enzyme/domain structure, starch binding site in domain C, catalytic mechanism starch substrate of different sources, e.g. wheat, wheat bran, rice bran starch substrate of different sources. Beta-amylase hydrolyzes alpha-1,4-linkage, raw starch granules from potato, wheat, rice and corn, with the granules from rice being the best substrate, no efficient hydrolysis of raw starch granules, very slow enzymic attack catalyzes the release of maltose from soluble starch. Malbranchea sulfureastarch substrate of different sources, e.g. wheat, wheat bran, rice bran 106.9% of the activity with amylose, soluble starch, amylose and amylopectin are the most suitable substrates, some activity against native starch, exo-hydrolase that releases beta-maltose from the non-reducing end of alpha-1,4-linked poly- and oligoglucans until the first alpha-1,6-branching point along the substrate molecule is encountered, beta-amylase should be a key enzyme in starch degradation during the germination of millet seeds, enzyme activity increases during days 1-4 of germination starch substrate of different sources, e.g. wheat, wheat bran, rice bran best substrate, pure and low quality starches, maize starch, tapioca starch maltose is the major end product, traces of maltooligosaccharides, no glucose as product. Beta-amylase is involved in starch degradation during mango ripening, which is clearly triggered by detachment from the mother-plant starch enzyme induction upon a cold shock at 4 °C leads to starch-dependent maltose accumulation, which might be required for protection of the photosynthetic electron transport chain, maltose influences the carbohydrate metabolism. Of the components of starch, amylopectin presents the great challenge to hydrolytic enzyme systems. This is due to residues involved in 1,6-glycosidic branch points which constitute about 4-6% of the glucose present. Most hydrolytic enzyme are specific for 1,4-glycosidic links yet the 1,6-glycosidic links must also cleaved for complete hydrolysis of amylopectin to glucose. Some of the most impressive recent exercises in the development of new enzymes have concerned debranching enzymes. It is necessary to hydrolyse starch in a wide variety of processes which may be condensed into two basic classes; Processes in which the starch hydrolysate is to be used by microbes or man and processes in which it is necessary to eliminate starch. In the former processes, such as glucose syrup production, starch is usually the major component of reaction mixtures, whereas in the latter processes, such as the processing of sugar cane juice, small amounts of starch which contaminate non-starchy materials are removed. Enzymes of various types are used in these processes. Although starches from diverse plants may be utilized, corn is the worlds most abundant source and provides most of the substrate used in the preparation of starch hydrolysates. There are three stages in the conversion of starch Gelatinisation, involving the dissolution of the nanogram-sized starch granules to form a viscous suspension; Liquefaction, involving the partial hydrolysis of the starch, with concomitant loss in viscosity; and Saccharification, involving the production of glucose and maltose by further hydrolysis. Galatinisation is achieved by heating starch with water, and occurs necessarily and naturally when starchy foods are cooked. Gelatinized starch is readily liquefied by partial hydrolysis with enzymes or acids are saccharified by further acidic or enzymic hydrolysis (Chaplin,2004). USES OF AMYLASE Amylase enzyme finds use in bread making and to break down complex sugars such as starch (found in flour) into simple sugars. Yeast then feeds on these simple sugars and converts it into the waste products of alcohol and CO2. This imparts flavour and causes the bread to rise. While Amylase enzymes are found naturally in yeast cells, it takes time for the yeast to produce enough of these enzymes to break down significant quantities of starch in the bread. This is the reason for long fermented doughs such as sour dough. Modern bread making techniques have included amylase enzymes (often in the form of malted barley) into bread improver thereby making the bread making process faster and more practical for commercial use. When used as a food additive, and may be derived from swine pancreas or mould mushroom. Bacilliary amylase is also used in clothing and dishwasher detergents to dissolve starches from fabrics and dishes. Workers in factories that work with amylase for any of the above uses are at increased risk of occupational asthma. 5-9% of bakers have a positive skin test, and a fourth to a third of bakers with breathing problems are hypersensitive to amylase. An inhibitor of alpha-amylase called phaseolamin has been tested as a potential diet aid. Blood serum amylase may be measured for purposes of medical diagnosis. A normal concentration is in the range 21-101 Mol/L. A higher than normal concentration may reflect one of several medical conditions, including acute inflammation of the pancreas, macroamylasemia, perforated peptic ulcer, and mumps. Amylase may be measured in other body fluids, including urine and peritoneal fluid. Two amylases are common to the baking industry, alpha-amylase and beta-amylase also known as alpha-1,4-glucan glucanohydrolase and alpha-1,4-glucan maltohydrolase. Amylases convert starch into sugar : the ÃŽÂ ±-amylase will cleave the starch randomly (the so called 1-4 bonds in the starch) while the ÃŽÂ ²-amylase can only chop off two sugar units at the time at the end of the starch chain. Normally there is enough ÃŽÂ ²-amylase present in the flour but sometimes addition of ÃŽÂ ±-amylase is needed. The ÃŽÂ ±-amylase will cut the starch into smaller units called dextrins and the more ÃŽÂ ±-amylase activity there is, the better for the ÃŽÂ ²-amylase because there are more extremities available. So the substrate for the ÃŽÂ ²-amylase is either starch or dextrins and the product is maltose. Alpha-amylase is an endoenzyme that attacks linkages within the molecular structure. It randomly cleaves starch chains at interior a-1,4-glycosidic linkages producing short chains of glucose molecules or dextrins. Beta-amylase is an exoenzyme and cleaves maltose units from the non-reducing end of the starch molecule. In order for these enzymes to function, the starch granule must be ruptured so that the individual starch molecules are available for enzymatic action. Depending upon their origin, alpha- and beta-amylases show differences in pH and temperature optima, thermostability, and other chemical stability. They do not require co-enzymes for activity, although alpha-amylase activity is enhanced by the presence of calcium. The pH optimum for alpha-amylase is 4.5 and it is inactivated at a pH of 3.3 to 4.0. This pH dependence decreases the efficacy of this enzyme in sour doughs. Beta-amylase is active across a much broader pH range, 4.5-9.2, with a pH optimum of 5.3. Alpha-amylase is relatively thermostable up to 70 °C, whereas beta-amylase loses about half of its activity at this temperature. Fungal amylase is the least temperature stable, followed by cereal amylase, while bacterial amylase is stable at higher temperatures. New intermediate stability enzymes have been developed that are active above the gelatinization temperature of starch (60 °C), but are totally inactivated at the later stages of baking (80-90 °C). The objective is to maximize the anti-staling effect without creating a gummy, sticky product. INHIBITORS OF AMYLASE ACTIVITY Amylase inhibitors are naturally present in many plants and protect the plant from pests by not allowing the insect to break down starch and gain energy from it. Plants may contain separate protease inhibitors as well or amylase inhibitors may play a dual role and also inhibit proteases. Protein amylase inhibitors as well as non-protein amylase inhibitors exist. Amylase inhibitors may be active against a wide variety of amylases or may be specific to certain insect amylases or mammalian amylases (Franco, et al 2002). Structure of Proteinaceous Amylase Inhibitors The determination of the structure of a complex between porcine pancreatic amylase  and a protein amylase inhibitor isolated from bean (Phaseolus vulgaris) showed interaction between the pancreatic amylase active site and the inhibitor. Conformational changes were observed in the pancreatic amylase upon the binding of the inhibitor. The inhibitor was found to be a dimer with a disaccharide attached to one of the amino acid residues.  (Bompard G., et al, 1996). Amylase Inhibitors in yam tuber (Shivaraj, et al,. 1979) reported that sweet potatoes do not contain amylase  inhibitors while (Rekha, et al,1999) reported the presence of amylase inhibitors in 79 of  the 100 varieties tested. Cultivar differences as well as isolation procedure could account for  these differing results. Before performing amylase inhibitor assays, Shivaraj and others  homogenized sweet potato with water, allowed the samples to sit for 1 hour, centrifuged the  samples, collected the supernatant, and then subjected the supernatant to heat treatment  (80 °C for 10 minutes) to destroy native amylases. Rekha and others homogenized yam tuber in a sodium phosphate buffer containing polyvinyl pyrrolidone and sodium chloride,  stored the samples in the refrigerator, centrifuged the samples, and then performed  trichloroacetic acid precipitation to remove native amylases. Rekha and others chose to use  TCA precipitation rather than the heat treatment Shivaraj found heat treatment to be in effective at destroying all native amylase activity. YAM Yam is the common name for some species in the genus Dioscorea (family Dioscoreaceae). These are perennial herbaceous vines cultivated for the consumption of their starchy tubers in Africa, Asia, Latin America and Oceania. There are many cultivars of yam. Yam (Dioscorea spp., Dioscoreaceae) is classified as monocotyledonous but is considered to be closely related to dicotyledonous plants as a second cotyledon remains undeveloped in the embryo (Lawton and Lawton, 1967). The storage organ is probably a swollen hypocotyl (Lawton and Lawton, 1969), but is often described as a swollen root. A number of species are grown widely in the humid tropics with D. rotundata and D. cayenensis being of most importance, followed by D. alata and D. esculenta (Akoroda, 1993). These are all of African or East Asian origin, with only the minor species D. trifida being of American origin (Brà ¼cher, 1989). The tubers contain about 1-3  % protein on a dry weight basis (Coursey, 1995). Yam is source of carbohydrate; carbohydrates are one of the three major food groups needed for proper nutrition. Amylase is the digestive enzyme needed to digest carbohydrates. Carbohydrates in food are an important and immediate source of energy for the body. Starch refers to carbohydrates found in plants (grains). Vegetables and fruits are a source of sugar and are broken down to sugar or glucose. Carbohydrates are present in at least small quantities in most food, but the chief sources are the sugars are the sugars and the sugars and the starches (Wright, 1993) Uses of yam Food Yams of African species must be cooked to be safely eaten, because various natural substances in raw yams can cause illness if consumed. (Excessive skin contact with uncooked yam fluids can cause the skin to itch. If this occurs, a quick cold bath will stop the itching.) Yam is consumed in various ways, but is usually boiled and eaten. This involves cutting yam into pieces, then peeling the skin, and boiling the starchy meat. This is usually consumed with palm oil (traditional way), or with other sauces. The boiled yam can also be pounded with a traditional mortar and pestle to create a thick starchy paste known as Pounded Yam. This is also eaten with traditional stews and sauces. Another method of consumption is to sun dry the raw yam pieces. When dry, the pieces turn a dark brown color. This is then milled to create a powder known as elubo in Nigeria. The brown powder can be prepared with boiling water to create a thick brown starchy paste known as amala. This is also consumed with the local stews and sauces. The most common cooking method in Western and Central Africa is cooked boiled yam. (Wikipedia 2003). In India this vegetable is also called Garadu. In central part of India people cut small slices of the vegetable, deep fry them, sprinkle lots of spices on it and eat as snacks. In southern part of India, it is eaten with fish curry and is a local favorite.(Wikipedia 2003) 1.2 TYPES OF YAM Dioscorea rotundata and D. cayenensis Dioscorea rotunda, the white yam, and Dioscorea cayenensis, the yellow yam, are native to Africa. They are the most important cultivated yams. In the past they were considered two separate species but most taxonomists now regard them as the same species. There are over 200 cultivated varieties between them. The Kokoro variety is important in making dried yam chips. They are large plants; the vines can be as long as 10 to 12 meters (35 to 40 feet). The tubers most often weigh about 2.5 to 5  kg (6 to 12 lbs) each but can weigh as much as 25  kg (60 lbs). After 7 to 12 months growth the tubers are harvested. In Africa most are pounded into a paste to make the traditional dish of pounded yam (Kay 1987). Dioscorea alata A piece of cake made with Ube (water yam).Dioscorea alata, called water yam, winged yam and purple yam, was first cultivated in Southeast Asia. Although not grown in the same quantities as the African yams, it has the largest distribution world-wide of any cultivated yam, being grown in Asia, the Pacific islands, Africa, and the West Indies (Mignouna 2003). In the United States it has become an invasive species in some Southern states. In the Philippines it is known as ube (or ubi) and is used as an ingredient in many sweet desserts. In Vietnam, it is called khoai mà ¡Ã‚ »Ã‚ ¡ and is used mainly as an ingredient for soup. In India, it is known as ratalu or violet yam. In Hawaii it is known as uhi. Uhi was brought to Hawaii by the early Polynesian settlers and became a major crop in the 1800s when the tubers were sold to visiting ships as an easily stored food supply for their voyages (White 2003). Dioscorea opposita Dioscorea opposita, Chinese yam, is native to China. The Chinese yam plant is somewhat smaller than the African, with the vines about 3 meters (10 feet) long. It is tolerant to frost and can be grown in much cooler conditions than other yams. It is now grown in China, Korea, and Japan. It was introduced to Europe in the 1800s when the potato crop there was falling victim to disease, and is still grown in France for the Asian food market. The tubers are harvested after about 6 months of growth. Some are eaten right after harvesting and some are used as ingredients for other dishes, including noodles, and for traditional medicines (Kay 1987).Air potato Dioscorea bulbifera Dioscorea bulbifera, the air potato, is found in both Africa and Asia, with slight differences between those found in each place. It is a large vine, 6 meters (20 ft) or more in length. It produces tubers; however the bulbils which grow at the base of its leaves are the more important food product. They are about the size of potatoes (hence the name air potato), weighing from 0.5 to 2  kg (1 to 5 lbs). Some varieties can be eaten raw while some require soaking or boiling for detoxification before eating. It is not grown much commercially since the flavor of other yams is preferred by most people. However it is popular in home vegetable gardens because it produces a crop after only four months of growth and continues producing for the life of the vine, as long as two years. Also the bulbils are easy to harvest and cook (Kay 1987). In 1905 the air potato was introduced to Florida and has since become an invasive species in much of the state. Its rapid growth crowds out native vegetation and is very difficult to remove since it can grow back from the tubers, and new vines can grow from the bulbils even after being cut down or burned (Schultz 1993). Dioscorea esculenta Dioscorea esculenta, the lesser yam, was one of the first yam species cultivated. It is native to Southeast Asia and is the third most commonly cultivated species there, although it is cultivated very little in other parts of the world. Its vines seldom reach more than 3 meters (10 feet) in length and the tubers are fairly small in most varieties. The tubers are eaten baked, boiled, or fried much like potatoes. Because of the small size of the tubers, mechanical cultivation is possible; which, along with its easy preparation and good flavor, could help the lesser yam to become more popular in the future (Kay 1987). Dioscorea trifida Dioscorea trifida, the cush-cush yam, is native to the Guyana region of South America and is the most important cultivated New World yam. Since they originated in tropical rain forest conditions their growth cycle is less related to seasonal changes than other yams. Because of their relative ease of cultivation and their good flavor they are considered to have a great potential for increased production (Kay 1987). Dioscorea dumetorum Dioscorea dumetorum, the bitter yam, is popular as a vegetable in parts of West Africa; one reason being that their cultivation requires less labor than other yams. The wild forms are very toxic and are sometimes used to poison animals when mixed with bait. It is said that they have also been used for criminal purposes (Kay 1987). 1.3 Nutritional value Yams are high in vitamin C, dietary fiber, vitamin B6, potassium, and manganese; while being low in saturated fat and sodium. Vitamin C, dietary fiber and vitamin B6 may all promote good health. Furthermore, a product that is high in potassium and low in sodium is likely to produce a good potassium-sodium balance in the human body, and so protect against osteoporosis and heart disease. Yam products generally have a lower glycemic index than potato products, which means that they will provide a more sustained form of energy, and give better protection against obesity and diabetes. Aim and objective The objective of this experiment is to extract the amylases mainly ÃŽÂ ²-amylase from yam tuber and determine the enzymatic activities of the enzymes. At the end of the experiment, the amylase extracted from yam tuber can be made use of in the industries like; the pharmaceutical, plastic and textile industries among others in place of barley commonly made used of. This is even favorable considering the land mass covered by yam and also large usefulness of the yam. CHAPTER TWO 2.0 MATERIALS AND METHODS 2.1 MATERIALS Yam (Dioscoreaceae esculenta),used was from Oja Oba Market in Iwo Osun State, Ethanol, Soluble starch, 3,5-dinitrosalicylic acid, sodium hydroxide, sodium potassium tertarate, Sephadex G200 was obtained from Pharmacia fine chemicals, Uppsala, Sweden, disodium hydrogen phosphate were products of British Drug House(BDH), poole England. The distilled water was obtained from the Department of Biochemistry, Obafemi Awolowo University, Ile Ife. 2.2 EQUIPMENTS Water incubator manufactured by Grant Instruments(Cambridge) Ltd, Weighing balance made in Switzerland, Centrifuge manufactured by microfield instruments England. Spectrophotometer. 2.2 Preparation of buffer and Reagents 2.2.1 preparation of 0.016 M sodium acetate buffer, pH 4.8 To prepare acetate buffer for, 73.10 g of sodium acetate was dissolved in 900 ml of distilled water, 4.2 ml of acetic acid was added and then made up to 1 Litre in volumetric flask. 2.2.2 Preparation of 2 N sodium hydroxide To 8 g of sodium hydroxide pellets was dissolved in 100ml of distilled water. 2.2.3 Preparation of colour reagent Dinitrosalicyclic acid colour reagent, was prepared by dissolving 1.0 g of 3,5-dinitrosalicyclic acid in 50 ml of distilled water. 30.0 g sodium potassium tartrate tetrahydrate was added slowly with 20 ml of 2N sodium hydroxide. It was diluted to 100 ml with distilled water. 2.2.4 Preparation of 1% starch Prepared by dissolving 1.0 g of soluble starch in 100 ml of 0.016 M sodium acetate buffer pH 4.8. It was boiled to dissolve and cooled, diluted to 100ml with distilled water. 2.3 Method The rate at which maltose is released from starch is measured by its ability to reduce 3,5-dinitrosalicylic acid according to Bernfold(1955) . One unit releases one micromole of ÃŽÂ ²-maltose per minutes at 25ËÅ ¡C and pH 4.8 under the specified conditions. 2.3.1 Mashing and Extraction of Enzymes With the use of mortar and pestle, 434.52 g of yam were ground and 400ml of homogenization buffer (i.e 0.016 M sodium acetate pH) was added and stirred, it was kept in a refrigerator for 1 hour with intermittent stirring at 10 minutes interval. It was then centrifuged at 4000 rpm for 10 minutes into components. Assay for protein and enzyme activity was then carried out after it has being stored in 70% of ammonium sulphate(212.4 g/L). 2.3.2 Purification Purification of ÃŽÂ ²-amylase: All enzymes purification steps were carried out at room temperature. Enzyme Precipitation: The crude extract was initially fractioned by 70% (v/v) ammonium sulphate. After centrifugation at 4000 rpm for 10 minutes, the precipitated pellets were collected and re-suspended in of cold buffer. The solution was dissolved in 0.016 M sodium acetate buffer of pH 4.8 and layered on a Sephadex G-200 Colum (1 x 40 cm). Fractions of 5 ml were collected. The fraction was monitored for protein at 280 nM. Elution was in 0.016 M sodium acetate buffer, pH4.8.

Lagrange Theorem

SAP transactions and reports Customer-specific transactions and reports Note Note It is possible that the origin of some objects cannot be identified clearly (either as SAP transactions and reports or customer-specific transactions and reports). In this case, the specified number of all transactions and reports is larger than the total of SAP and customer-specific transactions and reports. The possible deviation is due to the fact that only information for used objects is transferred. Objects in managed systems that exist there but are not used in the analyzed period are not transferred ND therefore have no designated origin.Information about customer objects that are not used is only displayed if you have selected the Get All Customer Objects option when you created the analysis. The following result types are displayed for each of the three named groups: Analyzed Not analyzed because, for example, there is no data for the client you specified when creating the analysis. Not analyze d in other clients The analysis fails with specified client due to missing data. Objects are used In other clients. You can base a new analysis with different clients, on this entry. Used.The objects are used in the analysis period and In the systems specified for the analysis. Not used (only affects customer-specific transactions and reports), If you have selected the Get All Customer Objects option when you created the analysis. Example Example You want to know how many transactions and reports were analyzed. The entry Is under All Transactions and Reports, Analyzed (Number). The transactions and reports of a result type, for example, the transactions and reports analyzed, are shown In a table. The following object properties are displayed: Object name, for example, transaction name such as SEES.Object type, for example, T for transaction or R for report. Object description, for example, Data Browser for transaction SEES. Origin, for example, S for SAP displayed in the All Transac tions and Reports overview Logical component, displayed for result type Analyzed, Not Analyzed or Not Analyzed in other Clients. SIDE, the system ID specified when creating the analysis; displayed for result type Used. System installation number; displayed for result type Not Used (customer objects). Threshold value, displayed for result type Analyzed, Not Analyzed or Not Analyzed in other Clients.Client specified when creating the analysis; displayed for result type Used. Result, for example, the number of dialog steps in a transaction Object description language You can copy objects of result types Used and Not Used to the rule database. Activities To open a table of transactions and reports, for example, a table of all the analyzed transactions and reports, click on the result type, for example on Analyzed. To copy an object to the rule database, click the result type, for example, Used to open the object table, highlight the object, and choose Add to Rule Database. Note NoteWhen copying the object to the rule database, you get a message informing you that a check step has been created or that the object is already in the rule database. To restrict the results to a specific system, open the input help for System and select a system. By default, the results refer to all systems you specified when creating the analysis. Note Note You can restrict the results by combining systems and time periods. You can choose another results overview tab: Graphical Overview Results in Analysis Structure Details of Analysis Error Statistics More Information Analysis Results Analysis Project Analyses Analysis Projects

Advertising and the construction of Violent White Masculinity Essay

The article â€Å"Advertising and the construction of Violent White Masculinity† points to the controversy of violence and media. It emphasizes that the mainstream debate about media and violence does not emphasis or analyze the most important aspect, namely gender. Katz states that the media illustrates a wrong image of individuals committing crime by calling them ‘youth’ crimes or ‘kids’ love. He believes however that the reason why crime is committed 90% by males is due to the masculinity emphasized in our society. He also mentions that there is lack of attention on criminals from majority groups such as Whites but an emphasis by crime conducted by minority groups. The article suggests that research has avoided to explicitly specify the relationship between masculinity and violent behavior by white men has been avoided in research.Katz believes to be able to illustrate some ways to display the hegemonic construction of masculinity that is present i n mainstream magazine ads and how these ads help to normalize violent male behavior. Katz suggests that one way violence is normalized is through the movie industry which shows white males engaging in violent actions without being depicted as villains. Moreover, they are regarded as heroes. The boom of the movie industry collided with the instability that was created economically for the White middle-class population. The heroes of the movie industry served as role models enabling the male white population to gain stability with masculine power through size and strength. According to Katz the physical body was the only way to obtain dominance and control for those who did not have economic resources to achieve manhood. Katz mentions another way our society tries to normalize violence in the white middle class, namely through advertisements. As Katz writes the ads are full of depictions of dangerous looking men. He points out the bidirectional relationship between advertisements and other socially accepted events such as sports and movies. Advertisements are filled with dangerous looking violent sports men or movie stars and on the other hand the movies or sports events display ads containing the same violent men, enhancing the impact these ads have on the white middle class. The other reason why the media exhibits the violent man is due to the decline of gender differences. Differences between males and females became more rigid. Men needed a way to differ from women, this was possible with becoming more identical with violent behavior. This is where violence becomes justified especially  against women as only a means to represent masculinity.Katz examines further the nature of ads regarding violence and masculinity. As he writes there are specific premises that are represented in ads about masculinity and violence. The first one is the notion that violence is genetically programmed male behavior. This notion is justified by ads showing historical events that were lead by powerful and violent and dangerous men. Some ads also show scenarios that involve violence against other ethnic groups or women. The second premise is concerned with the military and sports that are another source of masculinity. Uniformed soldiers and players are used in different kinds of ads. Using athletes or soldiers for these products, advertising creates the idea that using the advertised products will not decrease masculinity. The third point is the identification of muscularity with masculinity. Muscles are equated masculinity and power. Physical fitness becomes a major way for white men to keep their masculinity. Finally there is the notion of heroic actions being equal to violent masculinity. Movies play a very important role in illustrating this equation. One other significant point is that movies no only justify violence but also glorify violence.

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Essay on Is Addiction a Brain Disease - 2426 Words

Position Paper: Is Addiction a Brain Disease? Introduction Addiction, it is all around us, affecting people from all walks of life, it is not limited to certain social classes or lifestyles. It is found in every ethnic group, regardless of gender or age. It affects our neighbors, our friends, and our family either directly or indirectly. Although substances such as alcohol and illegal drugs are two of the most common addictions we hear about, there is a wide range of substances and even activities such as gambling and shopping. There is some debate whether addiction is a brain disease or a choice. This controversy has been going on for years and a decision whether or not addiction is a brain disease has yet to be reached.†¦show more content†¦If a person can make the choice to take drugs then they can also make the choice to stop taking drugs, especially during those times in between when they are not taking drugs. For a person to voluntarily continue taking drugs of their own accord then they should be held accountable for their addictive behavior and not use brain disease as an excuse. Further, circumstances such as economical, health, or personal issues resulting in a person’s choice to take drugs ironically can be the same reasons for them to either change their pattern, stop taking drugs, or get help versus the unavoidable, involuntary effect of a diseased brain. Additionally, those that choose to continue being addicted over a long period of time illustrates that by making that choice they are not helpless compared to a person who has a chronic disease. â€Å"Rather than being the inevitable, involuntary product of a diseased brain, these actions represent the essence of voluntariness. The addict’s behavior can be modified by knowledge of the consequences. Involuntary behavior cannot (Satel, Goodwin, 1998, pg. 5). As a result of that knowledge the addicted person is in control and can take be in charge of their actions to change the cycle and make a deliberate effort to change. 5 My Position: My position is optimistic onShow MoreRelatedIs Addiction A Brain Disease?1680 Words   |  7 Pages Cognitive Addiction Addiction is a Brain Associated Disorder that Allows the Body to Become Dependent on a Substance Oliver Sutton General Psychology 150-101 Professor Shawn Matthews 30 September 2014 Oliver Sutton Professor Shawn Matthews General Psychology 150-101 30 September 2014 Is addiction a brain disease? The definition of disease is a disorder in an organism that produces specific signs/symptoms that affects a specific location and isn’t a direct result of any kind ofRead MoreAddiction Is A Brain Disease Essay1119 Words   |  5 PagesLeshner and Addiction A prevailing belief in the field of addiction is that drug addiction is a â€Å"brain disease† characterized by â€Å"uncontrollable, compulsive drug craving, seeking, and use, even in the face of negative health and social consequences† (Leshner 2). In his article â€Å"Addiction Is a Brain Disease,† Alan I. Leshner claims that repeated use of drugs alters brain structure and function (1) and that once addicted, few are able to revert back to occasional use (2). Because of their â€Å"uncontrollable†Read MoreIs Addiction to Pornography a Brain Disease?1027 Words   |  4 PagesIs Addiction, Brain Disease? Pornography addiction and sex addiction are very alike sicknesses, arguably a sex addiction is not a disease or sickness however, and I feel as if it is. Walking around a sex addict you would probably never know about their disorder unless told otherwise. Many debates are about whether or not sexual addiction is an actual brain disease but just like drug addiction and alcohol addiction I classify it as the same. In a study taken in 2013 The Discovery Channel foundRead MoreAddiction Is A Chronic Brain Disease1757 Words   |  8 Pages Addiction is a chronic brain disease that often results in some sort of relapse. Addiction is characterized by inability to control drug use which results in problems with one’s behaviors and interpersonal relationships. This disease causes compulsive behaviors such as the need to use drugs despite the many harmful consequences that affect the addicted individual and those around him or her. Although for most people, the initial decision to use drugs is a one time lapse in judgement, the brain isRead MoreAddiction : Lack Of Will Power Or Brain Disease?1186 Words   |  5 Pages Addiction: Lack of Will Power or Brain Disease? Demonika M. Martin PS101 Introduction to Psychology Park University . I. Introduction Addiction is a disease that continues to fester and destroy individual’s lives. Once an individual is under the influence of drugs they no longer have control over their actions. Drug user’s brains are hijacked and taken on an explosive ride that begins with pleasure and ends in the damage of their brain. â€Å"Drug use is on the rise in this country andRead MoreDrug Addiction: A Brain Controlling Disease823 Words   |  3 Pagesdrug addiction has left a major impact on the brains of each user. Drug addiction is a relapsing brain disease that is characterized by brain malfunction that alters the way the users think and behave. Normal balancing Mechanisms go haywire, allowing the drug to control the brain. Drug addiction has been a major concern for scientist in recent years. With research drug addiction is categorized as a neuron-disease from its large impact on the brain. Drug addiction is a complex brain disease characterizedRead MoreThe Disease of Addiction Creates and Imbalance Between the Limbic and Frontal Lobes of the Brain1889 Words   |  8 PagesPatho-physiologists define addiction as a disease, thus affording to the chemical imbalance between the limbic and frontal lobes of the brain (Litowitz 2012). Psychologists have taken a broader approach, linking neuroscience with a binomial relationship to human behaviour in order to diagnose and challenge whether addiction is mind over matter, or indeed a neurological disorder (Burton, Westen, Kowalski, 2012). The following essay will firstly discuss three theories of psychology and how they eachRead MoreThe Problem Of Drug Addiction952 Words   |  4 Pagesthe eventual drug addiction is essentially involuntary. Drug addiction is a complex and chronic disease, a brain disease, which changes the way the brain functions. Drug addiction, much like chronic illnesses such as heart disease and diabetes , disrupts healthy, normal functioning organs. This has harmful consequences that are both preventable and treatable (Branch, 2011, pp.263-265). Drug addiction is considered a brain disease as drugs change the brain structure and how the brain operates. TheseRead MoreA Critical Look At Brain Disease1371 Words   |  6 PagesCritical Look at Brain Disease Addiction, such as drug or alcohol addiction, is explained by a Dr. Nora Volkow in a HBO series on addiction as a disease of the brain that translates into abnormal behavior. This disease is known as brain disease which refers to disruptions in the brain s motivational and reward circuitry that results from the cumulative effect of repeated use of certain substances. The documentary, Addiction, drove home the point that drug addiction is a brain disease that is aRead MoreAddiction as a Disease: Addiction is a term that has traditionally been used to refer to1400 Words   |  6 PagesAddiction as a Disease: Addiction is a term that has traditionally been used to refer to psychiatric syndrome that is caused by illicit drug use. Actually, addition is the only psychiatric condition whose symptoms are regarded as an illegal activity. In most cases, this term is described on the basis of drug use, which is the main focus of many research and treatment programs. Generally, drug addiction has significant negative effects on individuals using the drug and those around them such as

Black, White, Yellow And Brown - 909 Words

We are now in an age of colors with brown and yellow, not only the ancient divisions of black and white. Even though there are diverse people in the United States, it is totally absurd that if some people think America is a color-blind society. I used to think that colorblindness of black, white, yellow and brown was the only way to cut across the lines of racial equality issues. At that time, I was one of the supporters of color blind who think that â€Å"color blind† approach to the problems of racial inequality: The legal and political systems should simply ignore skin color and treat everyone the same† (172). Similarly, I believed that â€Å"Blind† refers treating individuals as equally as possible because people are blind to individual’s race, culture, and ethnicity. In this sense, colorblind society is another name of a society where everything is perfect and every individual is always happy, Utopia. On the other hand, I recently found out that color blindness might create or allow to persist entrenched discrimination while studying. In this respect, people put aside their differences. They are not only afraid of each other’s differences, but also do not want to recognize the difference. As the textbook stated, there are many differences between majorities and minorities. For example, white Americans are in a position of privilege. In other words, they have perceived the power of the privileged and the well off, unlike other racial minorities who are generally ignored by otherShow MoreRelatedThe Great Gatsby By F. Scott Fitzgerald951 Words   |  4 Pagesargument in the year of 2000 that Gatsby was a black man. Thompson explains his analysis and provides numerous facts from the â€Å"The Great Gatsby.† He shows how it is nearly impossible for an individual to say that Jay Gatsby was not a black man although he passes for a white man. Fitzgerald uses countless symbo ls throughout his novel, which can lead one to certainly agree with Thompson’s theory. Was Jay Gatsby a black man? Yes, Gatsby was almost certainly a black man. All of Thompson’s claims seem to beRead MoreEssay on Observations of Chemical Changes1398 Words   |  6 PagesAlso needed were, a piece of white paper, a piece of black paper, and the chemicals from the Observation of Chemical Changes Experiment Bag. For ever reaction, I wrote down the chemical combination, the well number, and my observations of the reactions against the white and dark backgrounds. Well#/Question | Chemicals | Reaction | A1/A | NaHCO3 amp; HCl - CO2 | The reaction with these chemicals, I had seen small white bubbles, both background white and black shows the same type of changeRead MoreThe Sound of a Hundred Feathers: The Symbolism of Richard Hooks Painting, Adoption of the Human Race748 Words   |  3 PagesRace, the main colors are a muted blue , a shading of yellow to dark brown ,black ,white and a secondary coloring of vibrant yellows and reds. The painting is that of Native Americans. In the Native American culture the color blue signifies wisdom and confidence. The chief is depicted in all blue. The color blue correlates with the role that is personified by the chief in the painting. The sky backdrop is black , black means strength. The black sky encases both the eclipsed moon and bolt of lightningRead More Color Blindness Essay752 Words   |  4 Pagescolors of the spectrum are able to be matched by mixtures of only three color sensitivities. 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The mustard yellow would be an acidic indicator and the royal blue a basicRead MoreExperiment to Prove Light is Needed for Photosynthesis798 Words   |  3 Pagesso that the light doesn’t get to that area if the leaf. I will the test the leaf for starch using iodine solution and the areas that got expose to sunlight will have starch and will turn a dark blue or black colour. The area that had the tinfoil in on it will have no starch and stay a yellow brown colour. Article How Photosynthesis works (article 1) Photosynthesis is divided into two parts as it is a very complex process. In the first stage which is the light dependent stage where lightRead MoreCharacteristics Of A Red Orange1643 Words   |  7 Pagesred to yellow, instead of having equal parts of red and yellow it would be two parts red and one part yellow. b. Red-Violet - This can be made when trying to create the secondary colors by adding more red to blue, instead of having equal parts of red and blue it would be two parts red and one part blue. c. Yellow-Orange - This can be made when trying to create the secondary colors by adding more yellow to red, instead of having equal parts of red and yellow it would be two parts yellow and oneRead MoreInvention Exhibit Each Area732 Words   |  3 Pagesand wall text was mostly white with boarders of the section’s theme color and the texts in black. Many of the signs and wall pieces had objects (gears, headphone, and cystoscopes) printed into paper behind the pexiglass. The base of the case hold the artifacts were metal and the covering for the cases was made of glass. The Hartford section used dark orange, brownish orange, and lighter orange. 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Blue dye amp; NaOCI. Record findings, then add 1 drop of HCI f. NaOCI amp; KI. Observe, then add 1 drop of starch. See starch turns black g.KI amp; Pb(NO3)2. Describe precipitate formed. h.NaOH amp; phenolphthalein. Observations

Meddle by Pink Floyd free essay sample

Meddle holds it’s own as the 6th album of English group, Pink Floyd. Arguably, Meddle is Floyd’s greatest album. The band packed so much sound in six songs that it’s almost impossible to hear the full capacity of the album through one sitting. It requires more, due it’s ocean deep depth. While this may seem intimidating to casual listeners at first, rest assured Meddle’s beauty comes from the complexity by being so seemingly simple at the first listen. Whether it be the incredible musicianship of the four Brits, or the watery effects of â€Å"Echoes†, Floyd created an album that’s a staple to any chill summer. The first song, â€Å"One of These Days† begins with windy effects that sound as if they were blowing off a coast itself. â€Å"One of These Days† has a driving bass that contrasts nicely to the dulcet, soft spoken quality of â€Å"A Pillow of Winds†. We will write a custom essay sample on Meddle by Pink Floyd or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Third track, â€Å"Fearless,† begins with heavy laden guitar chords in a mesmerizing riff you’ll be humming all day. The relaxedness of the tune and the decrescendo and crescendo combo of the thick piano sound and guitar chords, make it one of the standout tracks. In the background, you can hear screaming fans that sounds like a crowd cheering at a bullfight, when in fact, it’s the excitement of a crowd from the world cup soccer (football) game. The fourth song, â€Å"San Tropez†, is a music translation of comic strip Peanuts, if there ever was one. It’s almost as if you see Woodstock and Snoopy concocting up mischief in the backyard while Schroeder is composing on his baby grand. â€Å"San Tropez† brings a jazzy, head-bobbing vibe anybody needs after a long day at work. After â€Å"San Tropez†, a blues influenced howling song plays about a dog named Seamus (complete with actual dog howls). And at last, comes â€Å"Echoes†, the sixth and final track of Meddle. The twenty-three minute masterpiece is the perfect wrap-up to the album. For anyone unfamiliar with long songs, twenty-three minutes seems pretty daunting, with a dead spot nearly expected after the first three minutes. Instead, Floyd uses all twenty-three minutes to demonstrate superior musician mastery through drawn-out sounds layered with watery pings and slide guitar while the bass is played like waves rolling to the shoreline of a beach. Gilmour wails on guitar and teases you into a groove of funk for a while until about halfway through the song, screeches that resemble whale calls, come in. Floyd sends you directly into a musical climax of a ride and leaves you mellowed out long after the album stops playing. Sweet.