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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a 5-liter bioreactor yielding up to 12.5 U/ml phytase activity (1 g of PhyC-R/liter) in the cell-free culture medium (unpublished results) |
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2楼2010-11-12 08:15:27
wd519wd706(金币+8, 翻译EPI+1):感谢 2010-11-15 15:38:05
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芽孢杆菌的植酸酶是beta-螺旋桨式的植酸酶，在钙的存在下才有活性和保持稳定性，对于消化道为中性的动物来说，植酸酶的最佳pH范围为6.0- 9.0。芽孢杆菌在高温下，如60到95°C下的稳定性是作为动物饲料添加剂的另一项重要且有用的特性，因为在粒化的过程中需要蒸汽和高温。 在本篇文章中我们报道了从枯草杆菌VTT E-68013到植酸酶C基因的克隆和表达，使用巴斯德毕赤酵母为宿主。经过在摇瓶中48 h的甲醇浸入，挑选出的巴斯德毕赤酵母GS115-植酸酶C菌种产生并分泌出了植酸酶C的N-糖基化的形式，且细胞外的植酸酶活性为0.82 U/ml（71 mg的PhyC-R/升），比活为8.9 U/mg，代表了68%的细胞外蛋白质(0.1 g/升)。目前整个工艺并未完全优化，在无细胞培养环境中我们使用5升的生物反应器制得PhyC-R的植酸酶活性为12.5 U/ml（1 g of PhyC-R/升）（未见报道）。 由于不是生物专业的，有些可能不专业，见谅！ |
3楼2010-11-12 10:21:16
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2010-11-15 08:41
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