You are interested in examining the different kinetics of two isozymes of malate dehydrogenase (MDH). Isozymes are two similar enzymes that catalyze the same reaction but are from different genes or splice variants, giving them different properties. One of the MDH isozymes is located in the mitochondria, whereas the other is located in the cytoplasmic fraction. Your goal is to simply separate the two isozymes and test the samples in the partially purified state.

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Biology

5 July, 22:36

You are interested in examining the different kinetics of two isozymes of malate dehydrogenase (MDH). Isozymes are two similar enzymes that catalyze the same reaction but are from different genes or splice variants, giving them different properties. One of the MDH isozymes is located in the mitochondria, whereas the other is located in the cytoplasmic fraction. Your goal is to simply separate the two isozymes and test the samples in the partially purified state.

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Rory Salinas · Answer 1

Cell lysis

Explanation:

Since there are two different locations of the enzyme there are ways to separate them effectively. In order to achieve the purest possible native form the following can be done.

For the amounts in the MDH in the cytosol: the cells can be fractured and the centrifuged, All the aqueous material from the cytosol will be in the supernatent. This can be poured off and purified to the desired purity. Mitochonrtia MDH, all the mitochondria will be in the pellet. After the supernatent is poured of the pellet fraction that contains the mitochondrial MDH can be purified as the scientist sees fit.

Both types of enzymes can now be used in downstream processes