Procedures for the carbohydrate LAB Control experiment for m

Procedures for the \"carbohydrate LAB\" Control experiment for measuring glucose in an unknown solution 1. Get 10mL of distilled water in a 15mL conical tube and label it as \"CONTROL\" 2. Get 500HL of \"Unknown Glucose\" and pipette it into the CONTROL conical tube. 3. Take 50 HL of the \"Diluted unknown Glucose and pipette it in acentrifuge tube. 4. Label the centrifuge tube \"Control\". 5. Add 950 uL of Glucose Affinity Reagent to the \"Control centrifuge tube\" which contains the 50 HL of the \"Diluted Unknown Glucose\" already. 6. Mix the solution in the \"Control centrifuge tube\" manually for 10 secs. 7. Place the \"control centrifuge tube\" aside for 5 mins 8. Meanwhile set the spectrometer at 340 nm and blank it. 9. After 5 mins place the solution in a cuvette and place the cuvette in the spectrometer. 10. Record the absorbance value at 340 nm. 33s A Measuring glucose in an milk 1. Get 10mL of distilled water in a 15mLconical tube and label it as \"DILUTED MILK.\" 2. Get 500 uL of \"Milk\" and pipette it into the \"DILUTED MILK\" conical tube. c 3. Take 50 uL of the \"DILUTED MILK\" and pipette it in a centrifuge tube. 4. Label the centrifuge tube \"Milk 5. Add 950 uL of Glucose Affinity Reagent to the Milk centrifuge tube which contains the 50 HL of the diluted \"DILUTED MILK\" already. 6. Mix the solution in the \"Milk centrifuge tube\" manually for 10 secs. 7. Place the \"Milk centrifuge tube\" aside for 5 mins 8. After 5 mins place the solution in a cuvette a place the cuv meter. 9. Record the absorbance value at 340 nm. Measuring lactose in an milk 1. Take 50 HL of the \"DILUTED MILK\" and pipette it in a centrifuge tube. u 2. Label the centrifuge tube \"Milk Enzyme\". tube\" 3. Add 950 HL of \"REAGENT+B-galactosidase\" to the \"Milk t Enzyme centrifuge which contains the 50 uL of the diluted \"DILUTED MILK\" already. 10 secs. 4. Mix the solution in the \"Milk Enzyme centrifuge tube\" manually for 5. Place the \"Milk Enzyme centrifuge tube\" aside for 5 mins in the 6. After 5 mins place the solution in a cuvette and place the cuvette spectrometer. at 340 n 7. Record the absorbance value

Solution

1. There will not be any change in the product of the reaction if glucokinase is used instead of hexokinase as both the enzymes are capable of converting glucose into glucose-6-phosphate. The difference between Glucokinase and hexokinase is their Km value for glucose as glucokinase has lower affinity for glucose as compared to hexokinase which has very high affinity for glucose.

2. Enzymes needed for the reagent mixture to measure concentration of fructose 1-6 bisphosphate will be phosphohexose isomerase and phosphofructokinase.

4. Missing compound 1) Oxaloacetate

2) activator of Phosphofructokinase is ADP and Fructose 2,6 Bisphosphate(potent activator)

3. Question not very clear, But you can use all the enzymes of glycolysis that are used in the cycle till you get phosphoenol puruvate e.g you can start with hexokinase,phosphoglucoisomerase,phosphofructokinase, till the enzyme enolase that ultimately leads to formation of phosphoenolpyruvate.