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ananth ahil
ananth ahil's picture

Dear sir,
I am a JRF working in Polyhydroxyalkanoates production and purifiacation.Iam using p.putida for the production of MCL -PHA using glucose as a carbon source in nitrogen limited condition. So, intially I developed the medium Nutrient Rich Medium whch has rich nitrogen source, for promoting the biomass for my inoculum development. When I used to carried out the batch experiment, the OD reached 18, and the PHA yield percentagewas 9.3. When i opted for fed batch culltivation by exponential feeding  the od was not increased, it reached stationary phase,culture does not consume the glucose till the end of cultivation ,It was obseved by HPLC.we used 50 g/L of glucose and 0.66 g/L of ammoniumsulphate in fed batch fermentation.the media was designed by Metabolic flux analysis using GAMS .
Here i included the media composition which iam using for batch and fed batch fermentaions. 
Kindly do the needful
Nutrient Rich Medium  Composition :
Nutrient Broth  :10
Yeast Extract :15
Ammonium Sulphate:5

Basal Medium Composition :
(NH4)2SO4 - 0.66
Na2HPO4·12H2O - 7.30
KH2PO4 - 2.30
NaHCO3 - 0.30
CaCl2·2H2O - 0.01
MgSO4·7H2O - 0.25