Downstream Processing

Downstream Processing

 

 
Project description
Downstream Processing Assignment

Task Details <br />
A recombinant enzyme is to be manufactured in Escherichia coli. The enzyme helps clean up dead tissue, and therefore has a large potential market in the pharmaceutical industry. <br />
The enzyme is 14 kDaltons and has one disulphide bond. During fermentation, it can be formed as a correctly folded protein at low temperatures, or as an insoluble aggregate at high temperatures. Both forms are produced within the bacterial cell. The enzyme is robust and easy to refold if/when required. Experimental data has revealed that the enzyme binds selectively to arginine-sepharose chromatography media. <br />
You are required to design two processes to produce, extract and purify this active enzyme: one where the enzyme is produced as a soluble protein, and the other where it is produced as an insoluble aggregate. You must include process flow diagrams where appropriate, and justify your choice of process steps. Compare the two processes you have developed, and discuss the similarities between them, as well as the advantages and disadvantages of each. Based on this discussion, suggest (with justification) the most suitable process route for the production of this enzyme. <br />
Downstream Processing Assignment

Task Details
A recombinant enzyme is to be manufactured in Escherichia coli. The enzyme helps clean up dead tissue, and therefore has a large potential market in the pharmaceutical industry.
The enzyme is 14 kDaltons and has one disulphide bond. During fermentation, it can be formed as a correctly folded protein at low temperatures, or as an insoluble aggregate at high temperatures. Both forms are produced within the bacterial cell. The enzyme is robust and easy to refold if/when required. Experimental data has revealed that the enzyme binds selectively to arginine-sepharose chromatography media.
You are required to design two processes to produce, extract and purify this active enzyme: one where the enzyme is produced as a soluble protein, and the other where it is produced as an insoluble aggregate. You must include process flow diagrams where appropriate, and justify your choice of process steps. Compare the two processes you have developed, and discuss the similarities between them, as well as the advantages and disadvantages of each. Based on this discussion, suggest (with justification) the most suitable process route for the production of this enzyme.
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