Q1a. A biochemist is trying to improve the yield of the industrial enzyme invert
ID: 266812 • Letter: Q
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Q1a. A biochemist is trying to improve the yield of the industrial enzyme invertase that is used in the preparation of high fructose corn syrup. He has purified 2 different enzyme samples A and B. Outline an experiment to determine which sample has more enzyme activity. (4 m) Q1b. The stomach enzyme pepsin is used in the digestion of proteins. What is the expected optimum pH of a buffer for studying its activity? (2m) Q2. Vegetable oil was treated in 2 different ways. In the first reaction, it was subjected to the action of strong alkali (KOH) with boiling. In the second reaction, vegetable oil was treated with bile salts and the enzyme lipase. Draw out both reactions and explain the role of bile salts. (6m) Q3. You are given 6 standard carbohydrates: Starch, Glucose, Sucrose, Fructose, Ribose, Maltose and an unknown sample. The unknown gave a dark green black color with anthrone reagent, pale yellow color with lodine, a red orange cloudy precipitate with Barfoeds reagent, a blue color with slight red precipitate at the bottom of the tube with Benedicts reagent, was colorless with Seliwanoffs reagent and gave a brown layer with Bials reagent identify the unknown and provide a detailed explanation to justify your choice (6m) Q.4 The following samples: DNA, RNA, deoxyguanosine, adenosine, cytidine and ribose, were treated with the Dische reagent. State the principle of the Dische test and explain the expected results for each of the above samples. (6m)Explanation / Answer
The invertase enzyme can be quantified using DNS test. In this test, the invertase is allowed to convert sugar into glucose and fructose. These sugars reduce the DNS to give colourimetric readings.
1. Collect pure invertase enzyme and prepare multiple dilutions
2. Add sugar to each of the dilutions and incubate for 10 min.
3. Add DNS and incubate in hot water for 10 min.
4. Stop the reaction using H2SO4.
5. Observe the colour development in colourimetry.
6. Using this data, prepare a standard graph of invertase concentration v/s absorbance.
7. Use the two test invertase samples, and perform above experiment and using std graph measure the invertase concentration in samples.
2. usually stomach pH is around 1 - 2. So pepsin enzyme is best studied at pH round 2.
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