The nonoxidative glycolysis (NOG) pathway can achieve stoich…

The nonoxidative glycolysis (NOG) pathway can achieve stoichiometric carbon recovery for sugar-derived biosynthesis. Analyze the pathway figure below and answer the following questions: 1. In order to test whether this pathway can completely convert 1 F6P to 3 AcP, you were asked to try it in a test tube first. Please list the names of the enzymes in the labeled steps 2-4 that you need to purify to make this NOG pathway work.  (10 pts) 2. Which enzymatic steps are providing the thermodynamic driving force to make this NOG work? (5 pts) 3. You purified phosphoketolases (PK) (step 1) from three different organisms and tried them individually in the NOG reactions. After adding F6P and DHAP to the enzyme mixtures to start the reactions, you found all three PKs could drive the production of AcP. However, when you tested the PK activities, you found these PKs have different specificities toward F6P and X5P. Only one of the three has activities toward both F6P and X5P (F/XpK). The other two have only activities for either F6P (FpK) or X5P (XpK). Explain how were the other two PKs able to complete the NOG reactions? If there are any additional reactions that are not present in the figure to enable the NOG, please include these reactions in your answer. (20 pts)

You were tasked to analyze the oxidation rate of glucose by…

You were tasked to analyze the oxidation rate of glucose by the EMP glycolysis pathway in vitro (in a test tube). You purified all 10 enzymes in the EMP pathway and supplemented glucose, NAD+, and ADP in the test tube. When you analyzed the product pyruvate over a period of time, you found that its concentration was increasing but quickly reached plateau. In order to monitor the rate continuously, you added more glucose in the test tube but found no increase in pyruvate. Your advisor suggested adding two more enzymes in the test tube, a pyruvate decarboxylase (PDC) and an alcohol dehydrogenase (ADH), along with glucose. You were happy to find out that glucose can be continuously oxidized now as long as you supply glucose. 1. What could be the reason that pyruvate stopped increasing in the beginning even with the addition of glucose? (5 pts) 2. What product would you expect to detect when PDC and ADH were added to the test tube? (5 pts) 3. How did the addition of PDC and ADH help with the continuous oxidation of glucose? (5 pts)