State two thermodynamically nonideal conditions which could impact the AUC experimental analysis of proteins

Cells manage to synthesize a variety of seemingly thermodynamically unfavored molecules every day. Phosphorylated glucose is an example of one of these molecules. The direct phosphorylation reaction is depicted below. As indicated, the forward reaction is unfavorable with a positive standard free energy change:   glucose + phosphate⇄ glucose-6- phosphate + water                              ΔG°’ = +14 kJ/mol   Describe specifically a thermodynamically favorable route by which the cell can produce glucose-6- phosphate from glucose. Please note cells cannot change either the temperature or the pressure at which the reaction is occurring. Please help me understand by using steps. Thank you!

4. Below is a Michaelis-Menten plot for a wild-type (WT) and mutant (V105A) enzyme isolated from the bacterium Staphylococcus aureus. The enzyme is involved in carbohydrate metabolism and is a potential biocatalyst for the large-scale production of rare sugars. (8) mg V₂ μmol s1 1.50- 1.25- 1.00- 0.75- 0.50- 0.25- 0.00 0 100 200 300 [s], mM 400 500 WT (a) Estimate the KM and Vmax for the wild-type and mutant enzyme from the graph. V105A (b) Calculate the keat and keat/KM for the wild-type and mutant enzyme based on your estimated values in (a) if the total enzyme concentration is 0.5 µmol/mg. (c) Is the mutant enzyme a more or less efficient catalyst than the wild-type enzyme? Briefly explain.

Two-dimensional gel electrophoresis of proteins in a cell extract provides a qualitative way to compare proteins with respect to intra- cellular abundance. Describe a quantitative approach to the deter- mination of number of molecules of an enzyme per cell.

Where is the active site for this enzyme located?

If protein X produces a higher absorbance reading at pH 7 than it does at pH 9, what can you conclude?    Question 1 options:   A lesser amount of protein X was used in the reaction at pH 7 than at pH 9.   A greater amount of protein X was used in the reaction at pH 9 than at pH 7.   Protein X is a more efficient catalyst at pH 7 than it is at pH 9.   Protein X is a more efficient catalyst at pH 9 than it is at pH 7.

Why is it important to know the Km and Vmax values of an enzyme?