Validation of the model
• On different carbon sources (Glucose for example).
> > GrowthRate = 0.10;
> > CarbonIntake = �10.0;
> > model = changeRxnBounds(model,'BiomassReac_c_test_2',GrwothRate,'b');
> > model = changeRxnBounds(model,'GLC_tx_c',CarbonIntkRate,'b');
```
**> >** fba = MinOptmz

• Similarly perform the validation of model on nitrogen and sulfur sources i.e., reported in the literature. Draw the flux map to check the flow of carbon throughout the model.

Once the model has been developed and validated, write the model in either . mat, .sbml or .xlsx format for further analysis (**Figure 5C**).

#### **3.7 Target identification through essentiality analysis and gene knockout**

Developed model can be used for novel drug target identification by prioritizing essential genes through *in silico* gene knockout of single or double genes. Reaction deletion command lists out the reactions that help the pathogen model to cultivate too fast. On the other hand, gene knockout inhibit the particular gene of metabolome which act as chokepoint in pathogen. Consequently, it provokes the survival of pathogen that establishes the necessity of that gene as drug target for therapeutic use.

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