摘要
从118份样品中分离到1株产植酸酶的枯草芽孢杆菌(Bacillus subtilis WHNB 02),其发酵液经乙醇沉淀、硫酸铵分级沉淀及Sephadex G-100柱层析等步骤后分离纯化了该酶,纯化倍数约为31.5倍,回收率为13.0%.该酶为单体酶,SDS-PAGE测得的分子量约为43 KD,以植酸钠为底物的Km值为0.5 mmol/L,酶反应的最适温度为60℃,80℃作用10 min酶活保存61%,最适pH为7.0,在pH 6.0～10.0范围内稳定,酶活性及稳定性都需Ca2+存在.ED-TA,Mn2+,Ba2+(5 mmol/L)对酶活具有很大的抑制作用.
WHNB 02, an isolate of Bacillus suntilis, was obtained 118 samples. An extradellular phytase produced from it was purified to homogeneity by the steps of alcohol precipitation, （ NH4 ）,.SO4 fractionation and Sephadex G - 100 column chromatography. Purification of about 31.5 fold was achieved with an overall yield of 13.0%. The moklucular weight of the purified enzyme was estimated to be 43 kD on SDS - PAGE. The Km value for phytase was 0.5 mmol/L. The optimum temperature for phytase activity was 60℃ and about 61% of its original activity remained after incubation at 80℃ for 10 min. The optimum pH for the activity of the enzyme was 7.0 and it appeared stable in a pH range of 6.0 - 10.0. Calcium ions were required for its activity and stability. EDTA or metal ions such as Mn^2＋ and Ba^2＋ at 5mmol/L inhibited its activity.
出处
《西南农业大学学报（自然科学版）》
CSCD
北大核心
2005年第6期770-773,784,共5页
Journal of Southwest Agricultural University
基金
国家"十五"攻关项目资助课题(2002BA514A-12)
关键词
枯草芽孢杆菌
植酸酶
纯化
Bacillus suntilis
phytase
purification
