concentration of each enzyme and with [X] = 1 µM. Which curve corresponds to which enzyme? Explain.
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- Nicotiana tabacum cells are cultured to produce a polysaccharide gum. The reactor used is a stirred-tank reactor with an initial volume of 100 L. The maximum specific rate of growth of the culture is 0.18 d-1 and the yield coefficient of substrate in biomass is 0.5 gX/gS. The concentration of the limiting substrate in the medium is 3% (m/v). The reactor is inoculated with 1.5 g/L of cells and operated in batch until the substrate is exhausted, when the medium is fed with a constant flow rate of 8 L/d. The fed batch occurs in a quasi-steady state condition.a) Estimate the time of the batch step and the concentration of cells reached in this phase, considering exponential cell growth.b) The fed batch phase is carried out for a period of 40 days. What is the final concentration of cells in the reactor?c) The bioreactor is available for the process for 275 days a year, with an interval of 24 hours between each cultivation. What is the most advantageous operating mode for the process…The purification of cytochrome C begins with 1) yeast homogenization using a bead beater in the presence of BME (a reducing agent) and a protease inhibitor from approximately 900 grams of Baker’s yeast. Then, 2) insoluble cell contents were removed by centrifugation at 4,000 x g for approximately ten minutes. The ruptured cells (lysate) after centrifugation had a total volume of 0 mL and a 1.0 mL aliquot was set aside for further analysis. The following data was obtained from the 1.0 mL aliquot to quantify the protein amount and purity: The absorbance at 410 nm of the aliquot was 0.460 (1 cm pathlength). The absorbance at 595 nm from a 1.0 mL Bradford Assay solution that was diluted by 250-fold from the aliquot was 0.681 (1 cm pathlength). Using the information given, Calculate the total protein amount in mg from the absorbance at 595 nm. Calculate the cytochrome C amount in mg from the absorbance at 410 nm using Beer’s Law.An enzyme that catalyzes the reaction X ⇌ Y is isolated from two bacterial species. The enzymes have the same Vmax but different Km values for the substrate X. Enzyme A has a Km of 2.0 μM, and enzyme B has a Km of 0.5 μM. The plot below shows the kinetics of reactions carried out with the same concentration of each enzyme and with [X] = 1 μM. Which curve corresponds to which enzyme? Explain.
- If the volume of a Staphylococcus aureus cell is estimated at 0.5 μm3, how many cells could be accommodated, in principle, in 5 mL of saturated culture? (1 mL = 1 cm3). Show your calculations.Calculate the amount of GFP in grams produced in the 25-L fermentation used to generate cells for your homogenization lab. To do this problem, recall that for the homogenization step, E. coli cell paste was suspended in Tris buffer, the resulting cell suspension was homogenized. For this calculation, assume that the concentration of GFP in the clarified lysate is 2.0 mg GFP/mL solution. The concentration of cells in the 25-L fermentation was 8% by volume.in a clean, non-sterile 15 mL centrifuge tube, prepare a 2.0% yeast suspension by adding 0.06 g Saccharomyces cerevisiae to 3 mL yeast growing medium (56 mM glucose, 20 mM HEPES, pH 6.8). What percent of yeast suspension is left after a 1:10 dilution?
- Nutrient Agar (NA) is a general purpose medium used for the cultivation of a wide variety of non- fastidious microorganisms (Merck, 2000). Its ingredients are listed in Table 2. You are tasked to prepare 300ml of NA, determine the amount of ingredients and write in column 3 in Table 2. Show your calculations. *A sample calculation was made for you. Peptone = 300 ml x 0.5% = 1.5 g Table 2. Medium Composition of Nutrient Agar (NA) with the recommended proportions (Merck, 2019) Ingredients In % In Grams/300ml Peptone 0.5 *1.5 Meat extract 0.3 Agar 1.5 Distilled Water As neededStaphylococcal nuclease has a ΔΔG‡ of -84.1 kJ mol-1 at 25.0 °C. If the uncatalyzed rate is 0.630x10-13 µmol s-1, calculate the enzyme-catalyzed rate in µmol s-1. (Use R = 8.3145 J mol-1 K-1)You are cultivating Escherichia coli in a chemostat culture. The maximum specific growth rate (µmax) of E. coli that you can reach is known as 1.0 h-1 at your culture conditions. Describe what you would observe for each condition if you have the following settings: F (flow rate of the fresh medium into the bioreactor vessel) (L/h) V (Volume of the liquid culture in the bioreactor vessel) (L) a) 1 1 b) 5 4 c) 1 2
- You plot a reaction progress curve and find the equation of the line to be y = -0.0961x + 0.7142. Calculate the U, U/μL and U/mL of lysozyme activity in this fraction. The lysozyme activity is done identical to what is indicated in the lab manual.Inoculate 250-µL overnight cell culture into 50 ml LB medium (in a 250 ml flask). Shake vigorously at 37 °C to OD600~0.5-0.6 (usually it takes about 2-3 hours). Why should we use a larger flask during culture at this step? Why do we need to wait till this OD range is achieved?A bacterial culture is grown using either octadecane (C18H38) or pentachlorophenol (C6HCl5O) as the sole source of carbon and energy. The cell yield value is determined by dry weight analysis to be 1.49 for octadecane and 0.05 for pentachlorophenol. Using either octadecane or pentachlorophenol, please describe the steps taken (and the final result) showing what percentage of the substrate carbon will be found as cell mass and as CO2? Please show all calculations.