利用硫酸铵盐析缓冲液透析、聚乙二醇浓缩、DEAE-Sepharose FF窝子交换层析等方法,纯化了枯草芽孢杆菌WD-23 β-甘露聚糖酶,并研究了其酶学性质。结果表明:纯化的枯草芽孢杆菌WD-23 β-甘露聚糖酶的分子质量约为40 ku,酶的纯化倍数为14.1倍; 最适反应pH和pH稳定范围分别为5.6和5.0～7.0; 最适反应温度和温度稳定范围分别为55 ℃和40～70 ℃; Ca2+对该酶的促进作用最为明显,Li+的抑制作用最明显。
Abstract
The objective of this study was to purify β-mannase from Bacillus subtilis WD-23 and examine its enzymology properties. This study used the combining methods of ammonium sulfate, semi-permeable membrane dialysis, polyethylene glycol concentration, DEAE-Sepharose FF ion exchange chromatography, Sephadex G-75 gel filtration and SDS-PAGE gel electrophoresis to purify β-mannase. The result showed that the purified ratio of β-mannase of B. subtilis WD-23 was 14.1 by using the above methods, and the molecular mass was about 40 ku, the optimal pH and the stability range of pH were 5.6 and 5.0-7.0 respectively, and optimal temperature and the stability range of temperature were 55 ℃ and 40-70 ℃ separately, Ca2 + was the most obvious effect on the activation of the enzyme, on the contrary, the inhibitory effect of Li+ was the most obvious.
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基金
收稿日期:2013-10-26 修回日期:2014-03-06
基金项目:哈尔滨市科技局攻关项目(2008AA6B6BN025,GJ2011GG002112)
第一作者:张冰,硕士生; 向冰冰,硕士生。*通信作者:赵敏,教授。E-mail:82191513@163.com。
引文格式:张冰,向冰冰,崔岱宗,等. 枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质[J]. 南京林业大学学报:自然科学版,2014,38(3):88-92.
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