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The nucleophile in the first stage of the serine protease
mechanism is
a) H2O
b) the carboxyl of Asp 102
c) the hydroxyl of Ser 195
d) the imidazole of His 57
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I have given the protocol for the cyclodextrin glygosyl transferase assay: One ml of appropriately diluted enzyme sample was incubated at 60 °C for 15 min with 5 ml of 1% (w/v) gelatinized soluble starch in 50 mM, 7.0-pH Tris–HCl buffer. Reaction was terminated by boiling the reaction mixture for 3 min and reaction volume was made to 10 ml with distilled water. Two ml of above reaction mixture was withdrawn and mixed with 3 ml of Tris–HCl buffer, 5 ml of 125 mM Na2CO3, and 0.5 ml of phenolphthalein (25 mg phenolphthalein/100 ml absolute alcohol). Absorbance was measured at 550 nm. The percent decrease of sample was calculated with respect to control containing 5 ml of buffer, 5 ml of sodium carbonate and 0.5 ml of phenolphthalein. where Acontrol = absorbance of control and Atest = absorbance of sample. The amount of β-cyclodextrin (β-CD) produced was estimated from the standard graph of 0–500 μg/ml β-CD concentration against % decrease in absorbance. One unit of CGTase was defined as the amount of enzyme required to produce 1 μm of β-CD/min. Please can you suggest me the formula for the defination given in the last line
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