A16 Compound A is the substrate for two enzymes, E, and E2, involved in the synthe- sis of an amino acid and an antibiotic, respectively. Their reaction rates, r, and r2, at an arbitrary concentration of E, and E, were measured at different concen- trations of A. However, the data were not labeled properly (shown in Table P.4.1), and the student who did the measurement needs help to sort out which dataset is for E,. Determine the K and rmax tration of A. Which set of data is more likely to be for E, and which for E,, and why? for both enzymes, with respect to the concen- Table P.4.1 Reaction kinetics of a common substrate A for two enzymes. 3 4 5 Concentration of A (mM) 0.2 0.6 1.2 2 8 9 12 15 Reaction rate (r,) (mmol/L*min) 3.33 4.29 4.62 4.76 4.84 4.88 4.9 4.92 4.94 4.95 4.96 4.97 Reaction rate (r,) (mmol/L*min) 0.09 0.23 0.38 0.5 0.6 0.67 0.71 0.75 0.8 0.82 0.86 0.80

A set of biomolecules listed in the table at right are in solution at pH 6.8, when they are passed through the multi-step separation process shown below. The sample passes through an ultrafilter with MWCO of 50,000 (assume this filter gives a perfect separation of molecules above and below the MWCO), and the retentate then flows through an ion exchange column filled with beads that have a positive charge under the operating conditions (pks = 7.3). Inlet Biomolecule Mixture pH 6.8 Ultrafiltration MWCO 50,000 Permeate Retentate lon Exchange Column Biomolecule Molecular weight pl A 68,000 4.2 B 92,600 9.6 C 144,000 A-3. Which biomolecules (A, B, C, etc.) exit the process in the permeate stream? K C-3. Which biomolecules are retained inside the ion exchange column? D E F G H B-3. Which biomolecules exit the process after passing through the ion exchange column? 5,800 68,000 Packed with DEAE cellulose beads, which have positively charged groups below a pH of 7.3 156,000 45,000 16,000 None…

An enzyme contains an active site aspartic acid with a pKa = 5.0, whichacts as a general acid catalyst. On the accompanying template, draw thecurve of enzyme activity (reaction rate) versus pH for the enzyme (assumethat the protein is stably folded between pH 2–12 and that the active siteAsp is the only ionizable residue involved in catalysis). Briefly explain theshape of your curve.

Can you please help show how to find Kr?

A purified protein fraction has a total sample volume of 360 µL.  The sample has a corrected A280 of 0.484, and the blank corrected A280 was 0.052.  (Both values were measured with a path length of 1.00 cm.)  If 5.00 µL of the sample was used in a reaction, calculate the mass of protein in the reaction (in µg).