Lecture Notes 2

Lecture Notes 2 - Concept Question(s) on general bioprocess...

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Click to edit Master subtitle style 8/9/11 Concept Question(s) on general bioprocess
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8/9/11 Question setup, p 1 Consider a process that makes recombinant human streptokinase using E coli bacteria. At the end of the fermentation step, the streptokinase is intracellular (in the cytoplasm).
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8/9/11 Question setup, p 2 Fermenter Homogenizer Centrifuge Depth filter Ultrafiltration system Clarified lysate Purified streptokinase in PBS (Multiple purification steps: chromatography, etc.)
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8/9/11 Get pencil and paper handy for the CQ’s coming up…
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8/9/11 QUESTION 1 – working w enzyme Based on an enzymatic assay run at standard conditions, the concentration of streptokinase activity in the 2000 L of clarified lysate is 50 U/ml, and the total protein concentration is 20 mg/ml. (“U” refers to units of biological activity. 100 % pure streptokinase has a specific activity of 50 U/mg.) What is the specific activity of streptokinase in the clarified lysate? A ( red ): 0.4 U/mg B ( yellow ): 50 U/mg C ( green ): 1 x 105 U
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8/9/11 Answer to Q1 Streptokinase is an enzyme, as the suffix “ase” denotes. “Activity” in enzyme chemistry is an indirect measure of the mass of functioning enzyme in a sample based on the rate at which it can convert a substrate in an assay run at a standard set of conditions. Enzyme activity is proportional to enzyme mass. “Specific” means per unit mass of something*. The mass * For example, the typical units for the specific heat of a substance are calories/deg C per gram of substance .
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8/9/11 Answer to CQ1 – contd.
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Lecture Notes 2 - Concept Question(s) on general bioprocess...

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