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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medium (unpublished results)

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