| 枯草芽孢杆菌JNB001蛋白酶特性的研究 |
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| 引用本文： | 余全法,吴劲松,竺礼斌,王海霞,曲道峰,韩剑众.枯草芽孢杆菌JNB001蛋白酶特性的研究[J].中国畜牧兽医,2015,42(12):3261-3267. |
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| 作者姓名： | 余全法 吴劲松 竺礼斌 王海霞 曲道峰 韩剑众 |
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| 作者单位： | 1. 宁波市畜牧兽医局, 宁波 315012;2. 宁波市逸然畜牧有限公司, 宁波 315466;3. 浙江工商大学食品质量与安全系, 浙江省食品安全重点实验室, 杭州 310018 |
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| 基金项目： | 宁波市科技计划重大专项(2013C11033) |
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| 摘 要： | 为了解枯草芽孢杆菌JNB001发酵所产蛋白酶的特性,试验以蛋白酶活力为指标,研究了pH、温度等因素对酶促反应的影响,并通过水解度考察了粗酶液对各种动物蛋白的酶解效果。结果表明,酶反应最适pH为8.0,温度为45 ℃;粗酶液在pH 5.5~9.0的范围内较稳定,过碱则不利于酶液的保存。在45 ℃下热稳定性良好,超过50 ℃保温10 min后残余酶活已不足60%;金属离子Ca2+和Mg2+、还原剂DTT、表面活性剂Tween-80等能大幅度提高酶活力。而Fe2+、Cu2+、金属螯合剂EDTA、变性剂SDS等物质则会抑制蛋白酶活性;酶促反应的Km为6.43 mg/mL,Vmax为47.39 μg/min;针对不同蛋白底物,酶促反应较适pH为 7.0~9.0,温度为40~50 ℃。该粗酶液具有水解猪毛、猪蹄甲、脱毛牛皮、明胶等硬蛋白类物质的能力。上述试验结果可为枯草芽孢杆菌JNB001在病死畜禽无害化生物处理体系等方面的工业应用提供参考。
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| 关 键 词： | 蛋白酶 酶学性质 动物蛋白 酶解效果 |
| 收稿时间： | 2015-05-29 |
Study on Protease Properties of Bacillus subtilis JNB001 |
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| YU Quan-fa,WU Jin-song,ZHU Li-bin,WANG Hai-xia,QU Dao-feng,HAN Jian-zhong.Study on Protease Properties of Bacillus subtilis JNB001[J].China Animal Husbandry & Veterinary Medicine,2015,42(12):3261-3267. |
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| Authors: | YU Quan-fa WU Jin-song ZHU Li-bin WANG Hai-xia QU Dao-feng HAN Jian-zhong |
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| Institution: | 1. Ningbo Animal Husbandry and Veterinary Bureau, Ningbo 315012, China;2. Ningbo Yiran Animal Husbandry Co., Ltd., Ningbo 315466, China;3. Key Laboratory of Food Safety of Zhejiang, Department of Food Quality and Safety of Zhejiang Gongshang University, Hangzhou 310018, China |
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| Abstract: | This experiment was aimed to study the protease properties produced by Bacillus subtilis JNB001 via the index of protease activity to investigate the effects of pH,temperature and other factors on the enzymatic reaction,and the ability of the crude enzyme solution to hydrolyze various animal protein was measured by the degree of hydrolysis.The results showed that the most suitable pH of reaction was 8.0 and the temperature was 45 ℃.The crude enzyme solution in the range of pH 5.5 to 9.0 was relatively stable,while it was not suitable for the preservation of the enzyme solution when pH was over 9.0.At 45 ℃ it had good thermal stability when exceed 50 ℃ for 10 min, the residual activity had been less than 60%.Metal ions such as Ca2+ and Mg2+,reducing agent like DTT,surfactant as Tween-80,etc.could greatly improve the enzyme activity.On the opposite,Fe2+ and Cu2+ as well as other ions,metal chelator EDTA,denaturant SDS and other materials could inhibit the protease activity.The Km was 6.43 mg/mL,and Vmax was 47.39 μg/min.It had a range of reaction pH from 7.0 to 9.0 and temperature from 40 to 50 ℃ in the situation for different protein substrates.The crude enzyme solution could hydrolyze pig hair,pig nail,hair removal of cowhide,gelatin and other hard protein substances.The results provided the strong date support for the Bacillus subtilis JNB001 which could be used in dead livestock harmless biological treatment system and other aspects of industrial applications. |
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| Keywords: | protease enzyme properties animal protein enzymatic hydrolysis effect |
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