Isolation and Purification of Beta-Amylase from Soybeans
This paper is a detailed research protocol describing the methods and procedures necessary to isolate and purify beta-amylase from soybeans. Two different types of soybeans will be tested- green (immature) and brown (mature). Since the immature soybeans are sweeter, it is presumed that they have a higher beta-amylase activity. Mature beans, by contrast, are richer in protein, but are less sweet in flavor since there is less beta-amylase present to metabolize the starch into sugar. Therefore, the hypothesis for the experiment is: Green Soybeans have higher amylase activity compared to mature soybeans. This will be discovered by biochemical assays that measure the amylase activity in the soybeans. Following is a complete list of the steps in this process, including the preparation of special solutions, and detailed descriptions of the process to be undertaken. A complete list of reagents and chemicals is also presented here. 1.Gather materials and prepare solutions. 2.Isolation and Extraction Procedure Part 1 - Gather materials and prepare solutions. The first step is to acquire the soybeans. It is important to
Add the Maleic acid and EDTA to 800mL of distilled water and adjust pH tp 6.2 with 4M sodium hydroxide (16g/100mL). Add the sodium azide, dissolve and readjust the pH tp 6.2 if necessary. Adjust the volume to 1.0 L, then add BSA and dissolve. Store at 4 deg C. Remember that two doses will be necessary to test both types of soybeans. Dispense the Betamyl substrate solution into the bottom of glass test tubes (0.2mL each) and pre-incubate the contents at 40 deg C. for approximately 5 minutes. Also, pre-incubate the enzyme preparation at 40 degress C for approximately 5 minutes. TO each tube containing the Betamyl solution add 0.2mL of pre-equilibrated (and diluted) enzyme preparation directly to the bottom of the tube, and then incubate for at 40 deg C for exactly 10 minutes. After ten minutes has elapsed, add 3.0mL of stopping reagent to each tube and stir. Using the spectrophotometer (set at 410nm), read the absorbance of the reaction solutions and the reaction blank against distilled water. di-Sodium EDTA (Sigma ED2SS; 1 mM) 0.37g/L
Some common words found in the essay are:
Dispense Betamyl, Procedure Mill, Extraction Procedure, Introduction Background, Purification Method, Add Maleic, Green Soybeans, Predicted Results, Prepare Extraction, Reagent Top-pan, assay solution, 40 deg, enzyme preparation, amylase activity, extraction solution, approximately 5 minutes, maleic acid, mortar pestle, solution add, distilled water, ph tp, green soybeans amylase, ed2ss 1 mm, sigma ed2ss 1, materials prepare solutions,
Approximate Word count = 818
Approximate Pages = 3 (250 words per page double spaced)
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