To determine v max and km of Beta-galactosidase for lactose, the same amount of enzyme (1 ug per tube) was incubated with a series of lactose concentrations under conditions where product concentrations remained negligible. At each lactose concentration, the initial reaction velocity was determined by assaying for the amount of lactose remaining at the end of the assay.
Question: Calculate 1/v and 1/[s] for each entry. (Please note the equations used.)
Lactose concentration (m/M) (believe is [s])
1
2
4
8
16
32
Rate of lactose consumption (u mol/min) (believe is v)
10.0
16.7
25.0
33.3
40.0
44.4
(Please show me how to do aleast the first two. And the formulas used.)
Question: When graphing is 1/v on the y-axis and 1/[s] on the y axis?
To calculate 1/v and 1/[s] for each entry, we need to use the given lactose concentrations ([s]) and corresponding rates of lactose consumption (v).
To calculate 1/v, we take the reciprocal of the rate of lactose consumption. The formula for 1/v is:
1/v = 1 / (Rate of lactose consumption)
To calculate 1/[s], we take the reciprocal of the lactose concentration. The formula for 1/[s] is:
1/[s] = 1 / (Lactose concentration)
Let's calculate 1/v and 1/[s] for the first two entries:
For the first entry:
Lactose concentration ([s]) = 1 (m/M)
Rate of lactose consumption (v) = 10.0 (u mol/min)
1/v = 1 / 10.0 = 0.1
1/[s] = 1 / 1 = 1
For the second entry:
Lactose concentration ([s]) = 2 (m/M)
Rate of lactose consumption (v) = 16.7 (u mol/min)
1/v = 1 / 16.7 = 0.0599
1/[s] = 1 / 2 = 0.5
To graph 1/v and 1/[s], you should plot 1/v on the y-axis and 1/[s] on the x-axis. This is because in enzyme kinetics, the Lineweaver-Burk plot (also known as the double reciprocal plot) is commonly used. It is a linear representation of the Michaelis-Menten equation and makes it easier to determine parameters like Vmax and Km.
By plotting 1/v on the y-axis and 1/[s] on the x-axis, you will obtain a straight line. The slope of this line gives the value of Km, and the y-intercept gives the value of 1/Vmax.