You are required to isolate, purify and verify the purity of an enzyme from cytosol of a plant
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- Most enzymes do not operate at their biochemical optima in sidea cell. Explain why.To purify a given enzyme from a crude extract that contains it, fractional precipitations, exchange chromatography are successively tested ionic and exclusion chromatography, with the results shown in the following table: Volume of dissolution Protein concentration Enzymatic activity (ml) (mg/ml) (U/ml) a) Calculate the percentage of recovery of the enzyme after each of the manipulations to which it has been subjected. b) Indicate if it is necessary to continue purifying the enzyme or if it is expected to have reached its electrophoretic homogeneity.Briefly discuss the role of thiamine pyrophosphate in enzymatic reactions, using material from this chapter to illustrate your points.
- Idris has successfully extracted enzymatic proteins from the fish viscera (intestines and stomach). After homogenization and centrifugation, he managed to pool the crude enzyme extract. He is characterizing the enzymes. Please help Idris by answering the followingquestions:(a). How do I determine the enzyme activity? Please give the unit. (b) How do I get the specific activity of this enzyme? Please give the unit.Can you please help me answer the following question in three paragraphs and in your own words. 1. How do you determine the effects of substrate concentration on enzyme activity? (Do not write a procedure just give as many details as possible. Please make sure this is at least in three paragraphs and in your own words).Discuss how the equation for enzymatic reaction (given below) was demonstrated in the experimental results in Table 1. Use the appropriate color codes (-), (+), (++), (+++) to describe the expected results in Table 1 Table 1. Enzyme Action MIXTURE ENZYME ACTIVITY Water + catechol (tt A) Enzyme + catechol (tt B) Water + enzyme (tt C)
- Idris has successfully extracted enzymatic proteins from the fish viscera (intestines and stomach). After homogenization and centrifugation, he managed to pool the crude enzyme extract. He is characterizing the enzymes. Please help Idris by answering the followingquestions: (a) How do I run the experiment to find the kinetic properties of the enzyme, KMand Vmax? (b) The enzyme decreased in activity in the presence of NaCl. How do I find out if NaCl is a competitive or non-competitive inhibitor? Explain.True or False Immobilization improves the stability of the enzyme. EnaLne, has a half-life of 10 days in free solution, but under identical conditions of temperature, pH, and medium composition, the measured half-life of a packed column is 30 days. The enzyme is immobilized in a porous sphere 5 mm in diameter.Idris has successfully extracted enzymatic proteins from the fish viscera (intestines and stomach). After homogenization and centrifugation, he managed to pool the crude enzyme extract. He is characterizing the enzymes. Please help Idris by answering the following questions:(a) How do I determine the protein/enzyme concentration? Please give the unit.
- An enzyme catalysed reaction has a Km of 8 mM and a Vmax of 13 nM.s-1. Use the Michaelis-Menten equation to calculate the reaction velocity when the substrate concentration is 18 mM.Several factors contribute to enzyme catalysis. What arethey? Briefly explain the effect of each.Identify the enzymes in Table, whose catalytic effi ciencies are near the diff usion-controlled limit.