Dear all, Sorry for off topic discussion. i have a doubt for the enzyme kinetic.
[image: Inline image 2] Above is the reaction of my interest. it is a couple reaction. I want to determine the Km and Vmax for the E1 enzyme by the help of E2 enzyme by decreasing the amount of NADH (at 340 nm). if i don't know the optimum pH for E1. So is it ok, for publication point of view, to determine the Km and Vmax value of E1 enzyme at pH 7.5 ( a physiological pH) . Suppose if i determine the optimum pH of E1 by the help of E2 enzyme, that will be solely depend on the behaviour of E2 at different pH (if i am not wrong). Please suggest. Thanks in advance. -- WITH REGARDS Rohit Kumar Singh Lab. no. 430, P.I. Dr. S. Gourinath, School of Life Sciences, Jawaharlal Nehru University New Delhi -110067
