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- For the production of a secondary metabolic by Streptomyces coelicolor, a fed batch was performed. At the end of the single batch phase, the following conditions were reached in the reactor: V=10000L, cell concentration X=10g/L and product concentration P=0.1g/L. The feeding was then started with constant flow F= 200L/h, for 100h. Knowing that the substrate concentration in the feeding medium was SF= 80g/L and in the fermentation medium it was practically null, calculate: a) The final concentration of cells and productb) If the reactor were fed with a constant dilution rate (D), what should be the value of D used to reach the same cell concentration obtained in the situation with constant flow?μp= 0.01 g of product/ (g of cells.h)Y x/s = 0.15g cells/g of substrateSuppose that you were tasked with 4 cultures of a specific Bacteria Species in Luria Broth medium: (1) culture A – cells are in lag phase; (2) culture B – cell in log phase; (3) culture C – cells in stationary phase; and culture D – cells in decline phase. Imagine that you observe the growth rates of each culture in a fresh sterilized LB medium. Now, plot the outcome growth curves of cultures A, B, C and D in a single graph.A Fig. 2 Result showing the effect of different germicidal soap on the growth of bacteria Guide questions: 1. Based from figure 2 which soap is most effective? Explain 2. Which is least effective? Explain 3. Explain the result of disc D.
- Imagine you have been given a liquid culture of yeast with a starting concentration of 3.67 x 10' cells/ml and are asked to carry out the sample dilution process shown in the figure below. 100μl 100μl 100μl 100μl 100μl 0.9ml 0.9ml 0.9ml H2O H₂O 6.9ml 0.9ml H₂O H₂O H₂O Original 10-1 102 10-3 104 Culture 105 100μl 100μl 100μl Plate A Plate B Plate C a. How many colonies should have been present on Plate A in this example? - Answers must be whole numbers as partial colonies are not expected. b. Imagine you carried out the same dilution scheme shown in the figure above, but now, you do not know the concentration of the original culture. If you counted 163 colonies on Plate B, what is the concentration of cells/ml in the original culture?(a) Cell culture system can be maintained in several fermentation devices. Based on Figure Q2.1: (Sistem kultur sel boleh dikekalkan di dalam beberapa peranti penapaian. Berdasarkan Rajah Q2.1:1 OUTLET FOR SPENT MEDIUM LIGHT SOURCE RESERVOIR OF STERILE MEDIUM VALUE CONTROLLING FLOW OF MEDIUM PHOTO CELL Figure Q2.1: Laboratory setup for a cell culture system (Ali et al., 2018, Eur. J. Pharm. Med. Res.) [Rajah Q2.1: Penyusunan makmal untuk sistem kultur sel (Ali et al., 2018, Eur. J. Pharm. Med. Res.)]continuous disc stack centrifuge is operated at 5000 rpm for separation of tukers' yeast. At a feed rate of 60 min21, thirty of the cells are recovered. For operation at constant centrifuge speed, solids recovery is inversely proportional to the flow rate. (a) What flow rate is required to achieve 90% cell recovery if the centrifuge speed is maintained at 5000rpm thi What operating speed is required to achieve 90% recovery at a feed rate of 60 min211).
- You wish to centrifuge and pellet yeast cells using a centrifuge. The protocol says that you needto centrifuge the culture at 2,000xg for 10 minutes at room temperature. The rotor of yourcentrifuge has a maximum diameter of 25.4cm and the minimum diameter of 12.4cm. At whatrpm should you spin the centrifuge for pelleting the cells?Two flasks of E. coli are grown in batch culture in the same medium (2% glucose and amino acids; no nitrate) and at the same temperature (378C). Culture #1 is well aerated. Culture #2 is anoxic. After 16 hours the following observations are made: ■ Culture #1 has a high cell density; the cells appear to be in stationary phase, and the glucose level in the medium is reduced to 1.2%. ■ Culture #2 has a low cell density; the cells appear to be in logarithmic phase, although their doubling time is prolonged (over 1 hour). The glucose level is reduced to 0.2%. Why does culture #2 have so little glucose remaining relative to culture #1, even though culture #2 displayed slower growth and has less biomass?Experiment of batch fermentation of E. coli in a stirred tank bioreactor was carried out. 0.5g / L, 1g / L and 2g / L glucose were added on the bacteria, respectively. The absorbance values were measured by taking samples from the mixtures every 30 minutes. (Table 1). According to the values in the table, plot graph of cell- specific growth rate (μ / time) versus time.
- Devise a scheme to prepare a 10-9 dilution spread plate, using the least number of dilution tubes. Note: It is not practical to pipette any volume less than 0.1 ml and your equipment is similar to that in the Micro-lab, you only have 10 ml dilution tubes and a supply of sterile water Describe the growth patterns of each bacterial isolate that you observed in the nutrient agar deeps (shake cultures) and state what this says about their metabolism. Give definitions of these bacterial growth in relation to O2 E. coli: turbid growth throughout Micrococcus luteus:Growth on the top Clostridium sporogenes: growth on the bottomSydney Brenner isolated Salmonella typhimurium mutants that were implicated in the biosynthesis of tryptophan and would not grow on minimal medium. When these bacterial mutants were tested on minimal medium to which one of four compounds (indole glycerol phosphate, indole, anthranilic acid, and tryptophan) had been added, the growth responses shown in the following table were obtained. Mutant Minimal medium Anthranilic Indole glycerol acid Indole Tryptoph phosphate trp-1 trp-2 trp-3 trp-4 trp-6 trp-7 trp-8 trp-9 trp-10 trp-11 - Give the order of indole glycerol phosphate, indole, anthranilic acid, and tryptophan in a biochemical pathway leading to the synthesis of tryptophan. Indicate which step in the pathway is affected by each of the mutations. + 1 + 1 IISydney Brenner isolated Salmonella typhimurium mutants that were implicated in the biosynthesis of tryptophan and would not grow on minimal medium. When these bacterial mutants were tested on minimal medium to which one of four compounds (indole glycerol phosphate, indole, anthranilic acid, and tryptophan) had been added, the growth responses shown in the following table were obtained. Mutant Minimal medium Anthranilic acid Indole glycerol phosphate Indole Tryptophan trp-1 − − − − + trp-2 − − + + + trp-3 − − − + + trp-4 − − + + + trp-6 − − − − + trp-7 − − − − + trp-8 − + − − + trp-9 − − − − + trp-10 − − − − + trp-11 − − − − + Give the order of indole glycerol phosphate, indole, anthranilic acid, and tryptophan in a biochemical pathway leading to the synthesis of tryptophan. Indicate which step in the pathway is affected by each of the mutations.