POLINE MUTHONI MURIUKI

BSCN/2020/67229

MEDICAL BIOCHEMISTRY LAB REPORT

TITLE: DETERMINATION OF THE TCA CYCLE DEHYDROGENASE

AIM

To find the color change in tetrazolium depending on the levels of dehydrogenase enzyme
activity which drive the conversion of various intermediate with the help of co-factor and co-
enzyme.

BACKGROUND

The TCA cycle is a critical metabolic pathway which mediates energy generation. It plays an
integral role in the metabolism of lipids, carbohydrates and proteins.

TCA cycle hydrogenase drive the conversion of various intermediates with the help of co-
enzyme therapy yielding ATP.

MATERIALS

1. Oxaloacetate
2. Ketoglutarate
3. Malate
4. Distilled water
5. Liver
6. Hemogenate
7. Tetrazolium
8. Test tube

PROCEDURE

1. Label 5 test tubes as A, B, C, D, E

2. ADD 10 drops of oxaloacetate to testube A, 10 drops of ketaglutarate to test B, and 10
drops of Succinate to test tube C, 10 drops of malate to test tube D, and 10 drops of
distilled water to testube D.
3. Add 10 drops of liver homogenate in all testubes
Add 10 drops of tetrazolium to all testubes except testube F.
4. Gently swill and incubate for 10 minutes.

OBSERVATION
Intermediate observation
Oxaloacetate straw
Ketoglutarate straw
Succinate yellow
Malate orange
Water red

DISCUSSION
Tetrazolium acts as a proton acceptor thereby changing its color to red depending on the
product and the level of dehydrogenase activity.
Therefore, when the dehydrogenase is offered their respective substrate will produce
prospective products which can be detected by the color change of tetrazolium.
CONCLUSION
In oxaloacetate, there are low traces of dehydrogenase, in ketoglutarate has low traces of
dehydrogenase, succinate has moderate dehydrogenase, while malate and water have high
traces of dehydrogenase.

REFERENCES
Lininger’s Principle of Biochemistry by Nelson.