Bio-practical-3-report-amylase-activity-in-germinating-barley.pdf

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StuDocu is not sponsored or endorsed by any college or university Bio Practical 3 Report - Amylase activity in germinating barley Biology Ii (Monash University) Downloaded by Jackson Wong ([email protected]) lOMoARcPSD|5449659
Practical 3 1. Ten germinating seeds were dried and weighed before crushing them. 10ml of buffer was added slowly into the crushed seeds. The solution was filtered and the volume was measured and recorded. A diluted amylase extract was made by a five-fold dilution. A control extract was prepared by placing five ml of the diluted amylase extract into boiling water bath for 10 minutes. Boiled control extract was used to determine the effect of high temperature on the activity of amylase to hydrolyze starch as high temperature cause denaturation of enzyme. One drop of iodine was placed into 21 labelled wells. Reaction mixture was prepared from 5 ml of buffer, 1 ml of 0.5% starch solution and 1 ml of diluted amylase extract. A drop of reaction mixture was added immediately to the well labelled 0 and to sequential wells at every 1-minute interval until the achromic point was reached. This experiment was repeated by using boiled amylase extract. Amylase activity in germinating barley was calculated by measuring the amount of starch hydrolyzed per minute to reach achromic point and finding the mass of barley tissue used. It was calculated in the unit of mg starch hydrolyzed/min/g barley tissue. 2 ml of amylase reaction mixture and 2 ml of Benedict’s reagent were mixed and placed in the boiling water bath for 10 minutes. Any color change was observed and recorded to identify the presence of maltose. The experiment was repeated by using 10 dormant and 10 whole barley seedlings which include the use of a boiled control. If achromic point had been reached for the control, the experiment should be repeated by using additional buffer solution and less amylase extract while keeping the total volume at 7 ml. The experiment can also be repeated by

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