To express a yeast gene in E. coli, your task is to design a strategy to insert the yeast gene into the bacterial plasmid. Below is a map of the area of the yeast genome surrounding the gene in which you are interested.

 

The distance between each tick mark placed on the line above is 100 bases in length

Below are the enzymes you can use, with their specific cut sites shown 5’-XXXXXX-3’ 3’-XXXXXX-5’

 

The plasmid is 5,000 bases long and the two farthest restriction enzyme sites are 200 bases apart. The plasmid has an ampicillin resistance gene somewhere on the plasmid distal from the restriction cut sites.

                              

You do the digestion of the insert and the vector and then ligate the two digestions together. You then transform the ligation into bacteria and select for ampicillin resistance. You get three colonies on your transformation plate. You isolate plasmid from each one and cut each plasmid with the enzyme XbaI. You then run your three digestions on an agarose gel and see the following patterns of bands. Describe what each plasmid actually was that was contained in each of the three colonies.

 

What is the Colony 3’s plasmid is;

#Question id: 14309

#Section 5: Bioprocess Engineering and Process Biotechnology

Glucose is converted to ethanol by immobilized yeast cells entrapped in gel beads. The specific rate of ethanol production is: q_P = 0.2 g ethanol/g-cell-h. The effectiveness factor for an average bead is 0.8. Each bead contains 50 g/L of cells. The voids volume in the column is 40%. Assume growth is negligible (all glucose is converted into ethanol). The feed flow rate is F = 400 l/h and glucose concentration in the feed is S_0i = 150 g glucose/l. The diameter of the column is 1 m and the yield coefficient is about 0.49 g ethanol/g glucose. The column height is 4 m. What is the glucose conversion at the exit of the column?

#Question id: 14310

#Section 5: Bioprocess Engineering and Process Biotechnology

Glucose is converted to ethanol by immobilized yeast cells entrapped in gel beads. The specific rate of ethanol production is: q_P = 0.2 g ethanol/g-cell-h. The effectiveness factor for an average bead is 0.8. Each bead contains 50 g/L of cells. The voids volume in the column is 40%. Assume growth is negligible (all glucose is converted into ethanol). The feed flow rate is F = 400 l/h and glucose concentration in the feed is S_0i = 150 g glucose/l. The diameter of the column is 1 m and the yield coefficient is about 0.49 g ethanol/g glucose. The column height is 4 m. What is the ethanol concentration in the exit stream?

#Question id: 14311

#Section 5: Bioprocess Engineering and Process Biotechnology

Consider following fig, which applies to a fed-batch system.

Assume at t = 0, V = 100 l, X = 2 g/l, m = 1 h-1 , S0 = 4 g/l, and S = 0.01 g/l. V is increased at a constant rate such that dV/dt = 20 l/h = F (or flow rate) and X is constant at all times. What is µ at t = 5 h?

#Question id: 14312

#Section 5: Bioprocess Engineering and Process Biotechnology

An industrial waste-water stream is fed to a stirred-tank reactor continuously and the cells are recycled back to the reactor from the bottom of the sedimentation tank placed after the reactor. The following are given for the system: F = 100 l/h; S0 = 5000 mg/l; mm = 0.25 h-1 ; Ks = 200 mg/l; α (recycle ratio) = 0.6; C (cell concentration factor) = 2; Y ^M _X/S = 0.4. The effluent concentration is desired to be 100 mg/l.  Determine the required reactor volume.

#Question id: 14313

#Section 5: Bioprocess Engineering and Process Biotechnology

An industrial waste-water stream is fed to a stirred-tank reactor continuously and the cells are recycled back to the reactor from the bottom of the sedimentation tank placed after the reactor. The following are given for the system: F = 100 l/h; S0 = 5000 mg/l; mm = 0.25 h-1 ; Ks = 200 mg/l; α (recycle ratio) = 0.6; C (cell concentration factor) = 2; Y ^M _X/S = 0.4. The effluent concentration is desired to be 100 mg/l.  Determine the cell concentration in the reactor and in the recycle stream.

_____________

#Question id: 14314

#Section 5: Bioprocess Engineering and Process Biotechnology

An industrial waste-water stream is fed to a stirred-tank reactor continuously and the cells are recycled back to the reactor from the bottom of the sedimentation tank placed after the reactor. The following are given for the system: F = 100 l/h; S0 = 5000 mg/l; mm = 0.25 h^-1 ; Ks = 200 mg/l; α (recycle ratio) = 0.6; C (cell concentration factor) = 2; Y ^M _X/S = 0.4. The effluent concentration is desired to be 100 mg/l.  The effluent concentration is desired to be 100 mg/l. If the residence time is 2 h in the sedimentation tank, determine the volume of the sedimentation tank and cell concentration in the effluent of the sedimentation tank.