...coli Josh Brooks Department of Chemistry and Biochemistry, McMurry University, Abilene TX 79697 Abstract: In this experiment our goal was to analyze alkaline phosphatase from E. coli .In order to do this the purification of a protein from E. coli many steps were done through multiple weeks of assays. Finally, once the desired protein was pure a Bradford and a gel can be run to be sure that the protein was indeed purified. Introduction: In order to purify the protein many step are needed and the analyte should be evaluated by measuring the concentration and the activity of the protein. Physical and biological properties are used to help determine purity. These factors can be used to distinguish the analyte from other proteins. Located in the periplasmic space, the space between the cytoplasmic member and the outer membrane, alkaline phosphatase is cleaved and provides the cell with inorganic phosphate. To get a large enough sample of alkaline phosphatase the K12 mutant of E. coli is used. This mutant is unregulated in the cell because it lacks a way to control alkaline phosphatase production. Alkaline phosphatase is best used in acidic conditions and is very stable at high temperatures. By taking advantage of these two properties alkaline phosphatase can be isolated from the other proteins in E. coli. The overall goal is to reduce the total amount of protein but increase the specific activity of the desired protein. The experiment will also determine which particular method...
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...Date Sheet for Sciences General/ Core Courses/All Major Mid Term Examinations -Fall 2013 Semester Date & Day Sem 09:30 -11:00 Sec No. Room No. Teacher's Name Tooba Mohtsham Dr. Shahnaz Ch. Dr. Shahnaz Ch. Sem 11:30 -01:00 Sec No. Room No. Teacher's Name Sem Asifa Kayani Dr. Nikhat Khan 7 7 7 01:30 -03:00 Sec No. Room No. Teacher's Name 1 Introductory Biochemistry Introduction to Biotechnology Molecular Biotechnology A A A 9 35 4 Sci Y Sci Y Sci Y 3 3 Microbiology Electricity and Magnetism A A 56 31 NB-15 NB-8 Data Analysis & Report Writing A Data Analysis & Report Writing B Data Analysis & Report Writing C 33 NB-14 Farah Arif Munaza Bajwa Itrat Batool Naqvi 21-Oct-13 1 5 41 Main Lab NB-7 1 1 English-I English-I N K 25 44 SCI Y SCI Z Sadia Ghaznavi Nasreen Pashsa 3 Mathematics A 28 NB-36 Nighat Altaf 5 Molecular Physiology A 16 SCI 9 SCI 6 SCI 8 SCI 12 SCI 12 Tooba Mohtsham Asifa Kayani Saleha Mehboob Ayesha Aftab Gaitee Joshua 22-Oct-13 Basic Concepts of Environmental Sciences 24 5 A Data Handling and Atomic Spectroscopy 5 A 5 5 Electrical Instrumentation Human and Animal Behavior A A 9 9 12 7 Advanced Topics in Molecular BiologyA 7 7 Medical Biotechnology Plant Ecology A A 19 33 3 SCI 6 SCI R SCI 6 SCI 7 SCI 7 SCI 8 SCI8 Dr.Hooria Younas Dr. Amber Shehzadi Asifa Kayani Ayesha Roohi Saleha Mehboob Saima Mubeen Dr. Saleema...
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...Carbohydrates Carbohydrates are the sugars, starches and fibers found in fruits, grains, vegetables and milk products. They are called carbohydrates because, at the chemical level, they contain carbon, hydrogen and oxygen. Carbohydrates provide fuel for the central nervous system and energy for working muscles. Carbohydrates are classified as simple or complex. Simple carbohydrates contain just one or two sugars, such as fructose (fruits) and galactose (milk products). These single sugars are called monosaccharides. Carbs with two sugars — such as sucrose (table sugar), lactose (dairy) and maltose are called disaccharides. Complex carbohydrates, which are also called polysaccharides have three or more sugars. They are often referred to as starchy foods and include beans, lentils, potatoes, corn, whole-grain breads and cereals. Lipids Lipids are molecules that contain hydrocarbons and make up the building blocks of the structure and function of living cells. Examples of lipids include fats, oils, waxes, certain vitamins, hormones and most of the non-protein membrane of cells. Lipids are not soluble in water. They are non-polar and hydrophobic. Lipids contains a functional group including neutral fats, waxes, phospholipids, and glycolipids. The fatty acids with no carbon-carbon double bonds are called saturated. The ones that have two or more double bonds are called polyunsaturated. Proteins Proteins are large biomolecules, or macromolecules. They are made up of hundreds or...
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...There are four macromolecules that make up all living things. These macromolecules are proteins, carbohydrates, lipids, and nucleic acids. All of these macromolecules are formed from functional groups. The functional group hydroxyl is found in every macromolecule. All of these macromolecules except for lipids are hydrophilic. All of these macromolecules are broken down into monomers by hydrolysis reactions. Proteins have methyl groups, amino groups, and carboxyl groups. Proteins are polymers of amino acids. Proteins functions are determined by how they are folded. Proteins with a primary structure have no function. Proteins with a secondary structure are in the shape of an alpha helix but still have no function. Once a protein achieves tertiary structure that protein has a function. Quaternary structures in proteins are made of many motifs and have the most advanced functions of all the structures, many of them being enzymes due to their three dimensional structure. However, a protein can become unfolded, and once it does it cannot be reversed, this is called denaturation and makes the protein loose its functions. All proteins and polypeptides are links of amino acids held together by peptide bonds. Peptide bonds are formed between two molecules when the carboxyl group and the amino groups in two atoms react which causes dehydration synthesis. Carbohydrates have carbonyl groups, and are organic compounds every time due to the presence of a carbon atom and also have hydrocarbons...
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...compositional similarity with cells of higher organisms. This experiment makes use of these similarities to study individual macromolecular components found within all living cells. Through this experiment we will learn the basic sub-units that make up each of these macromolecules while also learning some of their important structural characteristics. This experiment will consist of two parts. The first of which will divide the yeast cells into three of its major macromolecular components: nucleic acids, proteins and polysaccharides. These components are large macromolecules that are quite unique in their composition, structure and function. However, they share a common feature as each macromolecule is composed of repeating subunits, characteristic of the macromolecule. The subunits are linked together by a bond between two adjacent subunits, formed by the loss of water (condensation). Thus, macromolecules can be broken down by the addition of water across the bond, in a process known as hydrolysis. This process was used in the experimental procedure to allow analysis of each individual macromolecule in its subunit form. Proteins are hydrolyzed into amino acids, nucleic acids are hydrolyzed into sugar, base and phosphate, and polysaccharides are broken down into simple sugars. In the second part of the experiment, the principle method of chromatography was used to analyze the macromolecules isolated in part one of the experiment. With this technique, individual molecular species were...
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...Biochemistry Empirical procedure for purifying Enzyme X: 1. To begin the purification process, mix blue green algae with an appropriate quantity of a buffer and triturate through use of a mechanical crushing process. Centrifuge at 4 degrees C for 10 min at 5,000 rpm. Next, determine whether the supernatant and the precipitate has the highest specific activity of the desired enzyme. The specific activity is the ratio of biochemical activity to the weight or volume of total protein present. The portion containing the greatest concentration of the desired enzyme would be kept for further fractionation and testing. Use of substances such as ammonium sulfate or polyethylene glycol that compete for water with the macromolecules could then be employed. This process, known as "salting out" the organic material from water by varying the solvent ratio or through doing another assay measuring the specific activity. Again, the portion containing the lesser amount would be discarded. Affinity chromatography would next be performed to identify the active enzyme based on the principal that enzymes attach to specific substrates and that specific receptors selectively bind the enzyme, impeding its passage in solution. Next, electrophoresis would be performed to help determine the purity of the compound. In electrophoresis, substances are first sorted by size and charge, and then by charge. There will be a separation of bands in the electrophoresis that corresponds to the...
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...Chemistry of Life – Lab Report The purpose of this lab exercise is to perform chemical tests on a variety of foods to identify the presence of any major molecules essential to life. After acquiring the results we will analyze them and we will be able to describe the general chemical content of those foods. In order to determine which specific major molecules were present in certain foods we conducted four different tests. The iodine test tested for starch by putting a few drops of iodine on a food substance. If the food turned a blue-black color then it was positive for starch. The Benedict’s test tested for the presence of sugar. In order to test you put 1 ml of solution to be tested and add an equal volume of Benedict’s reagent in a test tube and heat in a water bath for two minutes. If the solution goes from blue, to green-yellow, then to a red-brown color it means it is positive for sugar. The next test is the Biuret test and its tests for peptide bonds in proteins. To test you add a few drops of Biuret reagent to a food substance. If it turns a violet-purple color then it is positive for protein. The final test is the Grease Spot test for lipids. If there is a “grease spot” present after conducting this test, then the food tested positive for lipids. The first group of foods that were tested was the fruits and they consisted of apples, bananas, plantains, and avocados. The apple, banana, and plantain contained starch and sugar and this is correct because these two...
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...Devin Hoye Chemistry 1 9/8/12 4th Block 10Th Grade Lorenzo’s Oil Observations made in the beginning Lorenzo was a very normal , joyous kid. After a couple of days he started to have some behavioral problems in school. They really started to increase at home and at school. He is starting to go deaf. Each Day his condition worsens. The next day he is have problems with balance. There appears to be nothing wrong with him mentally. They have now diagnosed his disease . The name of this disease is rare , adrenlenscothy . Myelin another disease causes his brain to dysfunction. The doctor says he only have two years to live. His father researches the disease he finds out that at 20 Months he goes blind and at 24 months his death. He can barely walk at this point and needs assistance. 1 month after his kimo - therapy his hair has stared to fall out. His speech is started to be affected . His is fully aware that he is having seizures and he is in so much pain . He is fighting to his pain and his blood pressure goes down by 10 % at first then rises. By taking him off the fatty acid diet it affects him. The doctor asks Lorenzo asks him will he fine if other doctors are looking him at a meeting . Lorenzo at this point he can’t walk and needs help and when people thought he should be placed into special ed , he shows then he still can think for his self. . His parents tire-less work day and night to find solutions to there sons problem . Until they got a break through...
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...What is Biochemistry and why you should study it? Biochemistry or sometimes we called it as biological chemistry is defined as a scientific study of the chemistry of living organisms, especially the structure, the behavior of a living thing and the function of their chemical component such as proteins ,carbohydrates, lipids, and nucleic acids. Many of these molecules are complex molecules called polymers, which are made up of monomer subunits. Most people consider biochemistry to be same with molecular biology. Nowadays, biochemistry has become the root for understanding all biological processes. It has provided widely explanations for the causes of many diseases in humans, animals and plants. As a student, we should study it because it give to us many kind of knowledge on understanding the biological processes which are happen around us in every single minutes in our life. Since biochemistry is very important, we must study it to know how this biochemistry contributed for the sustainable of tomorrow in the main field of medicine, agriculture and industry. Biochemistry is applied in many health field such as dentistry, medicine and veterinary medicine. For example, in the field of medicine, biochemistry have contribute in the clinical study to maintain and to give a better and healthy life to the population of human all around the world. This have been done by the scientist on how they use biochemistry to diagnose and control the spreading of diseases, product a new...
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...Biochemistry Task 1 Sandra Clem 000448176 September 7, 2015 A. ( https://bnzm.files.wordpress.com/2010/09/dnareplication.jpg ) B. ( https://bnzm.files.wordpress.com/2010/09/dnareplication.jpg ) C. ( http://limbiclab.files.wordpress.com/2012/12/limbic_lab_levinandtijan2003transcriptionreg ulationandanimaldiversity.pdf ) D. The toxins that the death cap mushroom produce work against the enzyme RNA polymerase by stopping the transcription process. This in turn causes a domino effect as the mRNA will be stopped and no proteins will be regenerated from the cell. All of our tissues need protein to survive. If the death cap mushroom is ingested it starts this process in the GI tract. The liver and kidneys are filters for toxins and they try to filter the toxins. These toxins are prohibiting tissue repair by blocking RNA polymerase and no proteins can be made from mRNA this process causes liver and kidney failure and can cause death if not diagnosed early on. ( International Journal of Hepatology Volume 2012, http://dx.doi.org/10.1155/2012/487480) E. eferences R HudonMiller, S. (2012) Death cap mushrooms. Retrieved from https://wgu.hosted.panopto.com/Panopto/Pages/Viewer/Default.aspx?id=45c5aef2370e43ebad 9c40f073926331 . International Journal of Hepatology (Volume 2012), http://dx.doi.org/10.1155/2012/487480 ...
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...Biologically Important Molecules, Carbohydrates, Proteins, lipids and Nucleic Acids Tiffany Osbey Jackson State University Lab Report Authors: Tiffany Osbey Portia Mcmlertrey Jamia Ransome Darrius Bryant Abstract: Organic molecules are those primarily made up of carbon, hydrogen and oxygen. The common organic compounds of living organisms are carbohydrates, proteins, lipids, and nucleic acids. Each of these macromolecules (polymers) are made of smaller subunits (monomers). The bonds between these subunits are formed by dehydration synthesis. This process requires energy; a molecule of water is removed (dehydration) and a covalent bond is formed between the subunits (Fig.1). Breaking this bond is called hydrolysis; it requires the addition of a water molecule and releases energy. Each class of these macromolecules has different structures and properties. For example, lipids (composed of fatty acids) have many C-H bonds and relatively little oxygen, while proteins (composed of amino acids) have amino groups (-NH3+) and carboxyl (-COOH) groups. These characteristic subunits and chemical groups impart different properties to the macromolecules. For example, monosaccharide’s such as glucose are polar and soluble in water, whereas lipids are non-polar and insoluble in water. Objective: Perform lest to detect the presence of biology important carbohydrates, proteins lipids and nucleic acid. Explain the importance of a positive and a negative control in biochemical test. Use biochemical...
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...their monomers. • Summarize how polymers are made and broken down and how water participates in these reactions. • Describe the structure and functions of: o Simple sugars. o Nucleotides o Amino acids • Summarize the different forms and functions of complex carbohydrates. • Describe the possible levels of protein structure. • Summarize the fundamental property of all lipids • Illustrate the structure of triglycerides and phospholipids. Advanced learning objectives After the biomolecules lectures, students should be able to: • Predict what type of bond would be formed using electronegativity information. • Develop the critical thinking skills that allow you to evaluate scientific experiments that seek to explore how life started on earth. • Summarize why and where carbohydrates are commonly combined with other macromolecules. • Describe the relationship between functional groups and the molecules they are part of. • Compare and contrast the structure and functions of DNA and RNA. • Describe the four types of lipids. o Summarize and predict commonalities and differences in lipid structure and function • Describe how man-made fats are different from those found in nature and what their properties are. Be able to define, identify, and (where...
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...3.6 NOTES – MACROMOLECULES AS NUTRITION DO NOW: Identify the macromolecule: carbohydrate, protein, lipid, or nucleic acid. 1. | | 2. I am a macromolecule that is used as a source for short term energy. | | 3. | | 4. I am a macromolecule that is used to store genetic material. | | 5. | | 6. | | 7. | | 8. I am a macromolecule that is used for long term energy storage. | | 9. | | 10. I can be solid or liquid at room temperature. It all depends on the number of H-bonds in my “zigzag” lines | | 11. | | 12. I am a macromolecules with SO MANY different functions and SO MANY different shapes. | | 13. | | 14. The monomer for my macromolecule is a nucleotide. | | 15. | | NUTRITION LABELS AND DAILY DIETS The following macromolecules are essential to our diet: proteins, lipids, and carbohydrates. What percent of your daily diet does each macromolecule makeup? (% protein + % carbohydrates + % lipids= 100%) Guide to calories in macromolecules Macromolecule | Calories Per Gram | Protein | 4 | Carbohydrates | 4 | Lipids | 9 | Make a list of everything you ate yesterday. Identify the breakdown of proteins, carbohydrates and lipids. Try to come up with a good picture of which macromolecule makes up the majority of your diet. You can use the following website for nutritional information: http://nutritiondata.self.com/ or http://ndb.nal.usda.gov/ Daily Diet Breakdown...
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...substance. There are several different classifications of biomolecules that make up all the living things in this world. One of these classes is lipids, it is the only class that does not consist of polymers. There are three subcategories of lipids: triglycerides (fats), phospholipids, and steroids. All lipids share the same characteristic of being hydrophobic. Because they are hydrophobic, they separate from water and this makes it easy to test if a substance is made of lipids or not. In this experiment, we used ethanol to test...
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...Chapter 3 the Molecules of Life 1. Describe the causes and consequences of lactose intolerance. Lactose intolerance is the inability to digest lactose the main sugar found in milk. For people with lactose intolerance, the problem starts when lactose enters the small intestine. To absorb this sugar, digestive cells must produce a molecule called lactase. Lactase is an enzyme, a protein that helps drive chemical reactions. People with lactose intolerance produce insufficient amounts or a defective version of the enzyme. Lactose that is not broken down in the small intestine passes into the large intestine, where bacteria feed on it and belch out gaseous by-products, producing uncomfortable symptoms. 2. Explain why lactose intolerance has evolved differently in humans spread throughout the world. The enzyme lactase, like all proteins, is encoded by a DNA gene. Lactose-intolerant people have a defect in their lactase gene. A small change in a DNA nucleotide sequence can have a major effect on the production of a protein and the well-being of an organism. 3. Define macromolecules and list the three categories of macromolecules discussed in this chapter. Macromolecules are any very large complex molecule Three categories of macromolecules: 1) Carbohydrates 2) Proteins 3) Nucleric acids 4. Compare a dehydration reaction to hydrolysis. Dehydration reaction is a chemical reaction that removes a molecule of water. Cells link monomers together...
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