Hi,
I have just recently become acquainted with MCA and am interested in
applying it to a biotransformation system i am currently working on. The set
of reactions involved would be as follows
1. A -> B (enzyme E1)
2. B + NADH -> C + NAD (enzyme E2)
3. B + D -> E (enzyme E1)
4. D + NADH -> F + NAD (enzyme E2)
5. E -> G (enzyme E2)
A = pyruvate, B = acetaldehyde, C = ethanol, D = benzaldehyde, E = L-PAC,
F = benzyl alcohol, G = diol.
E1 = pyruvate decarboxylase, E2 = alcohol dehydrogenase.
E is the product of interest.
As is apparent from the above scheme, each enzyme handles more than one
reaction. I have at my disposal strains of yeast (S. cerevisiae) , with
varying levels of E1 and E2. Also inhibitors of E2 are also available.
Are there any examples of experimental work on systems where one enzyme
handles more than one step? I propose to calculate response coefficients
based solely on flux data ( from end product vs time profiles) and inhibitor
kinetics. However i am not too clear as to how i would be able to isolate the
effects of an inhibitor on a particular step when the enzymes involved handle
multiple steps. Also, would packages for MCA (metacon, gepasi etc ) be capable of handling such systems? ( I havent had a look at those yet, but would guess
that they would calculate FCC based on elasticities rather than direct
manipulation of flux data).
Any comments or assistance would be appreciated.
Thanx!
Santosh Noronha
noronha@ coe1.engr.umbc.edu
