BIO II Prac3.docx - Name Lim Connie ID 29274710 Practical 3...

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Name: Lim Connie ID: 29274710 Practical 3 Report Title: amylase activity in Dormant, Germinating and Seedling stages of barley Method: Amylase extract was prepared from germinating barley. First, dried and weighed 10 germinating barley seeds before crushing them. 10ml of buffer was added and continuously crushing and mixing for 3 minutes. Next, filtered the solution and measure the volume using measuring cylinder. A five-fold dilution was made by adding 20ml buffer and 5ml amylase extract to prepare a diluted amylase extract. A control was then done by adding 5ml of diluted amylase extract and was placed for ten minutes in the hot water bath. Hence, leave it for cooling at room temperature. Next, to calculate activity of amylase in germinating barley. Measure the amount of starch hydrolysed per minute to reach achromic point and mass of barley germinants used (in unit of mg starch hydrolysed/min/g barley tissue). In this session, on the ceramic plates, one drop of iodine was added into 21 wells. By adding 5ml of buffer solution and 1ml of 0.5% starch solution, a reaction mixture was prepared. Thus, add a drop of reaction mixture to a drop of iodine in the well. Furthermore, amylase reaction mixture was prepared by adding 1ml diluted amylase extract to reaction mixture. Immediately start with well 0 via add a drop of amylase reaction mixture to iodine. Add another drop of amylase reaction mixture to another well at every 1 minute interval each. Repeat this until achromic point was reached and record the time taken. Next, boiled control extract and repeat the experiment which is used to determine the influence of temperature on activity of amylase which hydrolyse starch. As at a higher temperature, it will cause hydrogen bonds holding enzyme amylase to break which will change the shape of enzyme and denatured.

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