I have absolutely no clue how to do this assignment Your supervisor at the Nutri

Question

I have absolutely no clue how to do this assignment
Your supervisor at the NutriZyme, Inc. has been charged with the task to establish a new manufacturing facility and design the processes for production of digestive enzyme supplements. The supplement is a mixture of protease (proteolytic enzyme), lipase, and amylase. He has assembled three task teams each assigned with the production of a specific enzyme and asked the teams each to submit a proposal detailing the selection of bacterial production strain, the selection of raw materials, the production process designs, the recovery and purification methods. Because of your prior trainings in industrial microbiology, you have been selected as the team leader responsible for the production of a- amylase. All enzymes produced should be in the form of dry powder ready to be pressed into pills or enclosed in gel caps. Your target yield is to produce 50kg of a-amylase from a 100,000L bioreactor You decide to seek assistant from the microbiologists on your team for the selection of bacterial strains, raw materials, and growth conditions. Your engineers are familiar with the design and control of production vessel and product recovery and purification methods Your microbiologists have made the initial progress report regarding the characteristics of 1 The a-amylase The target enzyme and its associated impurities have the following characteristics Net charge at pH 7.18 0 Ammonium sulfate | fraction (%) Proteins weight (kDa a-amylase 58 Impurity A 580 Impurity B 893 Impurity C 60 Impurity D 223 Impurity E 8 30 40 30 50 30 40 0 +2 -1

Explanation / Answer

1a . One can use a fixed angle rotor centrifuge to obtain the pellet and separate the culture supernatant.

1b to lyse the cells one can resuspend the cells in a particular buffer and use a high pressure homogenizer , upto 1000 bar . One can pass these bacterial cells twice for complete Lysis.

2. To obtain primary isolation one can use a particular concentration of ammonium sulfate to precipitate the protein of interest.

3. One can use ion exchange chromatography which relies on the principle of charge difference of the protein for separation. Care should be taken that conductivity should be less than 5mS/cm.

4. For final product isolation one can go for lyophilization so that water component goes off from the sample. Before that it is better to dialyse the product against water and then lyophilize.

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