Singh, SurenPermaul, KugenPuri, Adarsh KumarChanderman, Ashira2016-11-182016-11-182016663055http://hdl.handle.net/10321/1745Submitted in complete fulfillment for the Degree of Master of Applied Sciences in Biotechnology, Durban University of Technology, Durban, South Africa, 2016.A bacterial strain producing an extracellular phytase was identified as Enterobacter sp. ACSS. Optimization of process parameters using statistical methods such as Plackett-Burman design (PBD), the steepest ascent method, and response surface methodology (RSM) significantly improved phytase production by 4.6–fold in shake-flasks. In addition, an overall 1.9-fold increase in phytase production was attained in fed-batch fermentations in a 5 l laboratory fermenter, respectively. The purified 62 kDa phytase from Enterobacter sp. ACSS was active between 40 to 80°C and an acidic pH range of 2.0 to 6.0 with half-life of 693 and 577.5 min at 60°C and pH 2.0, respectively. Additionally, the enzyme is fairly stable with proteolytic enzymes under physiological conditions. It was activated by Ca+2, Mg+2 and Mn+2 while inhibition was caused by Zn+2, Cu+2, Fe+2, Pb+2, Co+2, Ba+2 and surfactants. The Km, Vmax and Kcat observed were 0.21 mM, 131.58 nmol mg-1s-1 and 1.64 × 103 s-1, respectively. The enzyme released inorganic phosphate from animal feed (4.0-6.62 mg/g of diet) and insoluble metal-phytates (45-219 µg/ml) and was effective in improving the characteristics of brown bread. Overall, this study shows that Enterobacter sp. ACSS has the potential to produce significant titres of a thermo- and acid-stable phytase and can be applied in dephytinizing animal feeds, and the baking industry.144 penPhytasesFeedsEnterobacterPhosphatesThesishttps://doi.org/10.51415/10321/1745