Phytase is used as a feed additive for degradation of antinutritional phytate, and the enzyme is desired to be highly thermostable for it to withstand feed formulation conditions. A Bacillus sp. MD2 showing phytase activity was isolated, and the phytase encoding gene was cloned and expressed in Escherichia coli. The recombinant phytase exhibited high stability at temperatures up to 100 degrees C. A higher enzyme activity was obtained when the gene expression was done in the presence of calcium chloride. Production of the enzyme by batch- and fed-batch cultivation in a bioreactor was studied. In batch cultivation, maintaining dissolved oxygen at 20-30% saturation and depleting inorganic phosphate below 1 mM prior to induction by IPTG resulted in over 10 U/ml phytase activity. For fed-batch cultivation, glucose concentration was maintained at 2-3 g/l, and the phytase expression was increased to 327 U/ml. Induction using lactose during fed-batch cultivation showed a lag phase of 4 h prior to an increase in the phytase activity to 71 U/ml during the same period as IPTG-induced production. Up to 90% of the total amount of expressed phytase leaked out from the E. coli cells in both IPTG- and lactose-induced fed-batch cultivations.

Oligochitosans (OCT) were prepared from chitosan (CTS) by gamma irradiationtechnique. The parameters affecting to the chitosan degradation were studied. And then, OCT nanoparticles wereformed using the method of tripolyphosphate (TPP) cross-linking. Effect of concentration and molecular weight of OCT, concentration of TPP on particle size of the formed OCT nanoparticles were also studied. The formation of OCT nanoparticles was verified by Fourier transform infrared (FT-IR) spectrometer and differential scanning calorimeter (DSC), the morphology was observed using scanning electron microscope (SEM), and the characteristics (particle size and zeta potential) of OCT nanoparticles were also studied. The effect of OCT nanoparticles on strawberry presevation was carried out using the coating method. Results showed that an increase in radiation dose resulted in a decrease of chitosan molecular weight. The OCT with molecular weight of approximately 7.7 kDa was obtained by the synergistic effect of hydrogen peroxide (5 %, v/v) and gamma ray at dose of 30 kGy. The smaller OCT nanoparticles was obtained with a lower molecular weight of OCT. The results of FTIR, DSC indicated the success in the formation of OCT nanoparticles with the particle size approximately 129.9 nm, with the spherical shape. The application of OCT nanoparticles on strawberry has prolonged the preservation times approximately 2.5 times higher comp