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The phytases from Bacillus species are beta-propeller phyta- ses requiring calcium for activity and stability and exhibiting an optimum pH between 6.0 and 9.0 and are suitable for animals with neutral digestive tracts. The stability of Bacillus phytases in a high temperature range, between 60 and 95°C, is another important and useful characteristic for their application as animal feed additives because the process of pelleting uses steam and high temperatures。 In this paper, we report the cloning and expression of the phyC gene from B. subtilis VTT E-68013 using P. pastoris as the host. After 48 h of methanol induction in shake flasks, the se- lected P. pastoris GS115-PhyC strain produced and secreted an N-glycosylated form of phytase C with an extracellular phytase activity of 0.82 U/ml (71 mg of PhyC-R/liter) and a specific activity of 8.9 U/mg protein, representing 68% of the total extracellular protein (0.1 g/liter). At present, while the biopro- cess has not yet been fully optimized, we are producing PhyC-R using a 5-liter bioreactor yielding up to 12.5 U/ml phytase activity (1 g of PhyC-R/liter) in the cell-free culture medium (unpublished results) |
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