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- topic: ISOLATION AND CHARACTERIZATION OF CARBOHYDRATES 1. Erythrose is an aldoterose. Describe the result when it is subjected to each of the following tests:a. Molisch’s Testb. Barfoed’s Testc. Benedict’s Testd. Seliwanoff’s Test2. Illustrate the reaction and identify the principle involved in the Nelson test.1. What is this lipid structure, give its role/ function, and enumerate its hydrolysis products or components. 2. Explain which hydrolyzed component will be positive in the Acrolein test? Ninhydrin test?4 A I. Refer to the figure below and answer the following questions: 45 5 55 6 65 PH 7 75 8.5 0 10 B 15 20 25 30 35 Temperature (°C) 40 45 Legend: Blue - wild-type ß-galactosidase; Red - mutant ß-galactosidase a. What is the optimum pH of wild type ß-galactosidase? b. What is the optimum temperature of mutant ß-galactosidase? c. Which enzyme has the greater activity at pH 7.2? d. Which enzyme has the greater activity at a temperature of 42.5°C? e. Which enzyme has greater activity if pH decreases from 7.5 to 6.4? f. Which enzyme has greater activity if temperature increases from 40°C to 41 °C? 50 55
- Based on this video https://www.youtube.com/watch?v=rKng5-ij6kQ Provide a schematic diagram for the Barfoed’s, test methodologies in determining the presence of carbohydrates. Also, give the basic principle for the test.Example of a Protein Purification Scheme: Purification of the Enzyme Xanthine Dehydrogenase from a Fungus Volume Total Total Specific Percent Fraction (mL) Protein (mg) Activity Activity Recovery 1. Crude extract 2. Salt precipitate 3. Ion-exchange chromatography |4. Molecular-sieve chromatography 5. Immunoaffinity chromatography 3,800 22,800 2,460 0.108 100 165 2,800 1,190 0.425 48 65 100 720 7.2 29 40 14.5 23 1.8 275 152.108 11 Calculate the specific activity of step#4. Note that percent recovery=% Yield.Based on this video https://www.youtube.com/watch?v=rKng5-ij6kQ Provide a schematic diagram for the Molish ’s test methodologies in determining the presence of carbohydrates. Also, give the basic principle for the test.
- C. Choosing the Proper Buffer Solution 1. Choosing the Proper Buffer Solution In Protein Precipitation, two liters of 5mM buffer solution with pH 5.2 is needed in the isolation of albumin. Which among the following buffer solution is best fitted for said purpose? Justify your answer. Buffer solutions pKa Acetate buffer 4.73 Tris- (hydroxymethy) aminomethane 8.08 Phosphate buffer 7.20 Discussion: 2. Preparation of the Chosen Buffer System Calculate and measure the amounts (in grams if solid and in mL if liquid) of weak acid and conjugate base needed to be able to prepare the chosen buffer system in part A above. Express your answer in useful units (that is, prepare it from practical amounts or concentrations of starting materials). D. Titration of an Amino acid Graph: titration of Glutamic acid Glu Glu Glu" Glu- COOH Co0 co0 Co0 H,N*-C -H H,N-C-H H,N*-c-H H,N -C -H CH2 CH2 CH, CH2 CH CH, CH, CH2 COOH соон co0 Co0 14 12 10 pK pH pkg Isoelectric point pk, 1.0 2.0 3.0 Equivalents of COH"…a. State the importance of using following reagents in SDS-PAGE. 1. Acrylamide 2. Bisacrylamide 3. Tetramethylethylelediamine 4. Glycerol 5. Ammonium persulfate b. Briefly describe the importance of two dimensional electrophoresis in protein separation?describe the structural features of protein transaminase/aminotransferases (primary, secondary, tertiary, or quaternary)? 2. identify the structural properties of transaminase/aminotransferases that are related to heat resistance or sensitivity and provided a plausible explanation?
- A. You are given the following carbohydrates: glucose, maltose, sucrose, and starch. Explain the results described below. a. A carbohydrate solution gave a negative result in the Benedict's and Barfoed's tests. When treated with hydrochloric acid and boiled for several minutes, the solution showed a positive result. Explain. b. A solution containing only one carbohydrate gave a Cu2O precipitate with Benedict's reagent. Which possible carbohydrates are present in the solution? c. Another sample of the solution from (b) failed to give a Cu2O precipitate with Barfoed's reagent within 5 min. Which carbohydrate is present in the solution?[C. Mucic Acid Test for Galactose and Lactose Galactose, on being oxidized with HNO3 forms mucic acid, an isomer of saccharic acid. Mucic is insoluble andforms characteristic sandy crystals which serve to identify galactose. Examine the crystals under the microscope. Describe the appearance of a few typical crystals.pls answer the following questions 1.Classify the following proteins according to their biological function: casein, albumin gluten, and myoglobin. 2.Which levels of protein structure organization are lost during hydrolysis and denaturation? 3.What is the Beer-Lambert's Law? Why is it relevant to the quantitative analysis of proteins? -What are the other protein components of milk aside from casein?