Team:Imperial College London/M1/Modelling

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==Model for drug enzyme kinetics==
==Model for drug enzyme kinetics==
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The drugs we are manufacturing are enzymes, so in order to detect them in their assay, analysis of how they interact with substrates is required.  
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The protein of interest is an enzyme.  It will bind to specific substrates and increase the rate of their conversion into products.  Therefore, by monitoring either the substrate concentration or the product concentration, we can indirectly see the activity of the enzyme.  This is quantified by measuring by enzyme activity, in the [https://2009.igem.org/Team:Imperial_College_London/Wetlab/Protocols/cellulase assay] and [https://2009.igem.org/Team:Imperial_College_London/Wetlab/Protocols/PAH PAH assay].  Enzyme activity is the rate of substrate utilisation /product formation per unit time.
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Revision as of 13:44, 12 September 2009

Contents

Introduction

This module consists of producing the drug protein of interest. In order to describe the function of the module, two models have been developed.

Model for protein drug production:

Based on the Genetic circuit, a LacI-IPTG inducible promoter is responsible for kickstarting the production of the drug.

  • In the absence of IPTG, LacI represses the production of the drug (Cellulase or PAH)
  • When IPTG is introduced, the LacI repressing pathway is “de-repressed”, and some output protein is produced.

Our goals

The modelling aims to provide an overview and better understanding of the M1 system’s function by:

  • Characterizing the system.
  • Modeling to account for several factors that may reduce/hinder the production of the protein drug such as:
    • Lac promoter leakiness
    • IPTG toxicity
    • Stability of output protein

The conclusions from the models should help people from the Wetlab to plan their experiments and take into account these limitations. This module is an integral part of the design, as large-scale commercialization of the drug of interest depends on finding the optimal conditions for protein production.

Model for drug enzyme kinetics

The protein of interest is an enzyme. It will bind to specific substrates and increase the rate of their conversion into products. Therefore, by monitoring either the substrate concentration or the product concentration, we can indirectly see the activity of the enzyme. This is quantified by measuring by enzyme activity, in the assay and PAH assay. Enzyme activity is the rate of substrate utilisation /product formation per unit time.



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