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Forty grams of powdered Jueyen chili was mixed with 30.00 mL cold of Tris-HC extracting buffer to isolate its crude protein content. Then, 20.00 mL aliquot of the crude protein extract was partially purified using ammonium sulfate precipitation at 60% to 80% saturation. The protein content of the crude and partially purified protein extracts was determined using a colorimetric Bradford assay with Bovine Serum Albumin (BSA) as standard. The results of the experiment were as follows: Absorbance of 2 mL standard BSA solutions Concentration of the Absorbance at 595 nm Absorbance of crude and partially purified protein extracts Sample Absorbance at

Forty grams of powdered Jueyen chili was mixed with 30.00 mL cold of Tris-HC extracting buffer to isolate its crude protein content. Then, 20.00 mL aliquot of the crude protein extract was partially purified using ammonium sulfate precipitation at 60% to 80% saturation. The protein content of the crude and partially purified protein extracts was determined using a colorimetric Bradford assay with Bovine Serum Albumin (BSA) as standard. The results of the experiment were as follows: Absorbance of 2 mL standard BSA solutions Concentration of the Absorbance at 595 nm Absorbance of crude and partially purified protein extracts Sample Absorbance at

Chemistry
Chemistry
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ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
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Forty grams of powdered Jueyen chili was mixed with 30.00 mL cold of Tris-HCl extracting buffer to isolate its crude protein content. Then, 20.00 mL aliquot of the crude protein extract was partially purified using ammonium sulfate precipitation at 60% to 80% saturation. The protein content of the crude and partially purified protein extracts was determined using a colorimetric Bradford assay with Bovine Serum Albumin (BSA) as standard. The results of the experiment were as follows: Absorbance of 2 mL standard BSA solutions Concentration of the Absorbance at 595 nm standard BSA solution, pg/mL Absorbance of crude and partially purified protein extracts Sample Absorbance at 595 nm |Undiluted Crude Protein Extract 1:5 Dilution Crude Protein Extract 1:10 Dilution Crude Protein Extract Undiluted Partially Purified Protein Extract 1:5 Dilution Partially Purified Protein Extract 1:10 Dilution Partially Purified Protein Extract 0.00 0.150 1.496 62.50 0.166 0.832 250.00 0.200 0.286 375.00 0.224 2.300 500.00 0.252 1.205 750.00 0.298 0.340 1000.00 0.351
Transcribed Image Text:Forty grams of powdered Jueyen chili was mixed with 30.00 mL cold of Tris-HCl extracting buffer to isolate its crude protein content. Then, 20.00 mL aliquot of the crude protein extract was partially purified using ammonium sulfate precipitation at 60% to 80% saturation. The protein content of the crude and partially purified protein extracts was determined using a colorimetric Bradford assay with Bovine Serum Albumin (BSA) as standard. The results of the experiment were as follows: Absorbance of 2 mL standard BSA solutions Concentration of the Absorbance at 595 nm standard BSA solution, pg/mL Absorbance of crude and partially purified protein extracts Sample Absorbance at 595 nm |Undiluted Crude Protein Extract 1:5 Dilution Crude Protein Extract 1:10 Dilution Crude Protein Extract Undiluted Partially Purified Protein Extract 1:5 Dilution Partially Purified Protein Extract 1:10 Dilution Partially Purified Protein Extract 0.00 0.150 1.496 62.50 0.166 0.832 250.00 0.200 0.286 375.00 0.224 2.300 500.00 0.252 1.205 750.00 0.298 0.340 1000.00 0.351
Table 1. Ammonium sulfate saturation table (grams per 1L sample). Final concentration of (NH4)2SO4, % saturation 50 60 70 80 10 20 30 40 90 100 Grams of solid (NH4):SO4 to be added to 1 L of solution 0 55 113 176 242 314 390| 472| 561 657 761 10 57 117 182 251 325 405 491 584 685 20 59 121 188 260 337 421 511 609 30 61 126 195 270 351 438 533 40 63 130 202 281 365 457 50 65 135 210 292 381 60 67 140 219 305 70 70 146 228 80 73 152 90 76 1. Determine the amount of powdered ammonium sulfate needed in bringing the solution from (a) 0% to 60% saturation and (b) 60% to 80% saturation assuming that the volume remains constant. 2. Calculate for the protein concentration (µg/mL) of the crude and partially purified protein extracts. 3. Based on these observations, was the partial purification of the target protein successful or not? Justify. 4. Calculate the total protein content (g) of the Jueyen chili sample using the calculated crude protein extract concentration. Initial concentration of (NH4)2SO4, % saturation
Transcribed Image Text:Table 1. Ammonium sulfate saturation table (grams per 1L sample). Final concentration of (NH4)2SO4, % saturation 50 60 70 80 10 20 30 40 90 100 Grams of solid (NH4):SO4 to be added to 1 L of solution 0 55 113 176 242 314 390| 472| 561 657 761 10 57 117 182 251 325 405 491 584 685 20 59 121 188 260 337 421 511 609 30 61 126 195 270 351 438 533 40 63 130 202 281 365 457 50 65 135 210 292 381 60 67 140 219 305 70 70 146 228 80 73 152 90 76 1. Determine the amount of powdered ammonium sulfate needed in bringing the solution from (a) 0% to 60% saturation and (b) 60% to 80% saturation assuming that the volume remains constant. 2. Calculate for the protein concentration (µg/mL) of the crude and partially purified protein extracts. 3. Based on these observations, was the partial purification of the target protein successful or not? Justify. 4. Calculate the total protein content (g) of the Jueyen chili sample using the calculated crude protein extract concentration. Initial concentration of (NH4)2SO4, % saturation
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