Tacrolimus Experiment

Tacrolimus is specific T-cell inhibitor and potent immunosuppressant agent [no]. Tacrolimus (FK506 or Fujimycin) is a 23-membered polyketide macrolide with immunosuppressant activity that was discovered in culture broth of the soil bacterium Streptomyces tsukubaensis in 1984 []. The immunosuppressant tacrolimus was initially discovered in fermentation broth of Streptomyces tsukubaensis, which was isolated from Japan soil of the Tsukuba region. Most of the initial studies performed with streptomyces tsukubaensis had been focused on strain improvement and development of fermentation procedures at industrial level []. In initial studies Fujisawa scientists isolate a tacrolimus producing strain of streptomyces tsukubeansis, no 9993 and established

Response surface methodology is a group of techniques that are used to study the relations between one or measured dependent factors (responses) and several input (independent) factors [27]. The effect of concentration of the three selected variables, ammonium sulphate, yeast extract and 1,2 propylene glycol was studied by this method. The concentration ranges selected for the three factors are listed in (Table 4). All other factors (glucose 10 g/L, glycine 0.2%, temperature 28 °C, initial pH of medium 7.0, agitation rate 200 rpm, inoculum volume 2%) were kept constant. To calculate optimum values of selected three factors, a set of 20 experiments was generated using a 23 full factorial CCD, with six replicates at the centre point, was employed to fit a second order polynomial model. In each experiment the 250 ml flask contained 50 ml production medium and three factors namely ammonium sulphate, yeast extract and propylene glycol, were varied in their levels according to the design of the software. All other factors were kept constant (glucose 1 %, glycine 0.2 %, temperature 28 °C, pH 7.0, agitation rate 200 rpm). All the flasks were incubated at 28 °C in Kuhner incubator shaker. Samples were taken at 9

The Placket-Burman experimental design matrix set up for the screening of main factors for tacrolimus production (continued)
Run
No Propranol (%) CaCO3 (g/L) FeSO4 (g/L) MgSO4 (g/L) Tacrolimus production
(mg/L)
Experimental