Team:METU Turkey/circuit.html

• P450 Monooxgenase Production and its Activity:

Overview:

The aim of modeling P450 monooxygenase production is to predict the amount and the activity of P450 enzyme which will be produced from our circuit . The model was created mainly by designing ODEs and enzyme kinetics equations .Then; these equations were solved by the help of MATLAB software.

As seen in the circuit, CYP6G1 gene , which will produce P450, is synthesized by using B. subtilis specific constituve promoter Pveg. fCPR  gene, used for activation of CYP6G1 in B. subtilis, is also transcribed with same promoter. These two genes has transcribed with SacB genes, after translation which helps to export these proteins through outside of the cell, into the bee gut. [1]  To be able know, how much enzyme is produced in a given time and how much imidacloprid is converted into nontoxic components by P450,  model of this circuit was required.

Differential Equations :

PARAMETERS

N             Plasmid Copy Number

P             Promoter Strength

amrna     Degradation rate of mRNA

K             Dissociation Constant

N             Hill Coefficient

T             Translation Rate

D             Protein Degradation Rate

Enzyme Kinetics for P450 Monooxygenase:

To be able to estimate the rate of the P450 reaction with imidacloprid,  we used Michaelis-Menten Kinetics.  The harmful imidacloprid concentration for honey bees is reported as 1200 µg/l [2], so in our model we assumed that the imidacloprid concentration in bee gut as 1200 µg/l. And P450 monooxgenase will be produced from our circuit and directly secreted into bee gut where imidacloprid is found.

 According to Michaelis-Menten Kinetics;