摘要
以枯草芽胞杆菌LD-8547菌株发酵液为材料,提取分离溶栓酶,研究了几种有机溶剂及变性剂对枯草杆菌溶栓酶活性的影响.结果表明,甲醇对溶栓酶表现为抑制作用,在10%～15%浓度范围时,其抑制作用最强;乙醇在5%～15%浓度范围内对酶活有激活作用,但随浓度的增大则转变为较强的抑制作用;正丁醇和异丙醇对酶活力则表现出强烈的抑制作用;丙三醇对酶活力有轻微的抑制作用,随着浓度增加抑制作用逐渐增强;异戊醇在5%～15%浓度范围内有一定的激活作用,但当浓度大于15%时则表现为抑制作用.醛类对酶活也有较强的抑制作用;丙酮、DMSO、DMF、SDS和异氰硫酸胍对酶活均有不同程度的抑制作用;脲浓度<0.01mmol/L时略有激活作用,随浓度增大转变为抑制作用.实验同时对EDTA在不同浓度和不同时间内对酶活力的影响进行了研究,结果表明,浓度<2mmol/L的EDTA基本不影响酶活力,但随着浓度的增大,其抑制作用明显增大.本实验还以牛血为材料,对酶液进行了体外抗血凝和溶血栓实验,结果表明,该酶具有抗血凝和溶血栓效果.
The fibrinolytic enzyme was extracted and purified from fermented broth of Bacillus subtilis LD-8547. The effects of different organic solvents and denaturants on the enzyme activity were investigated. The results indicated that methanol inhibited the enzyme activity, and the inhibitive effect reached the peak at the concentration of 10%-15%. Alcohol increased the enzyme activity a little at the concentration of 5%-15%, but inhibited it at high concentration. Butanol and isopropyl alcohol showed a strong inhibition on the enzyme activity. Glycerol had a light inhibitive effect on the enzyme activity, and its inhibitive effect gradually increased when its concentration became higher. Isoamyl alcohol could slightly affect the enzyme activity at the concentration of 5%-15%. But when the concentration was greater than 15%, it showed an inhibitive effect. Aldehydes had a relatively strong inhibitive effect on the enzyme activity. Acetone, DMSO, DMF, SDS and guanidine sulfate could inhibit the activity at different degrees. Urea showed a weak inhibitive effect at a concentration lower than 0.01 mmol/L, but its effect became stronger with the increasing of its concentration. The effects of EDTA on the enzyme activity at different concentrations and different times were also studied, indicating that EDTA almost had no effect on the enzyme activity when its concentration was lower than 2 mmol/L. However, with the increasing of its concentration, the inhibitive effect of EDTA significantly increased. At the same time, the anticoagulant and thrombolytic functions of this enzyme in vitro were studied by using cow blood, showing that this enzyme had both anticoagulant and thrombolytic effects. Fig 2, Ref 13
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2008年第6期825-829,共5页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(Nos.30500325
30771400)
国家"863"计划项目(No.2007AA10Z430)
霍英东教育基金会高等院校青年教师基金项目(No.111032)
福建省高校新世纪优秀人才支持计划资助~~
关键词
溶栓酶
有机溶剂
变性剂
溶血栓
fibrinolytic enzyme
organic solvent
denaturant
thrombolytic
