| 一株产耐高温蛋白酶蜡样芽胞杆菌的分子鉴定和酶学性质研究 |
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| 引用本文： | 佘新松,邵建扬,孙春巧,方茹月,储雅琪,江治萍,代雨婷,柏晓辉.一株产耐高温蛋白酶蜡样芽胞杆菌的分子鉴定和酶学性质研究[J].核农学报,2020,34(8):1698-1704. |
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| 作者姓名： | 佘新松 邵建扬 孙春巧 方茹月 储雅琪 江治萍 代雨婷 柏晓辉 |
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| 作者单位： | 1 黄山学院生命与环境科学学院,安徽 黄山 245041; 2 绩溪县林业局,安徽 宣城 245300 |
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| 基金项目： | 安徽省高等学校自然科学研究重点项目;草原局科技中心项目;安徽省自然科学基金;国家林业;国家级大学生创新创业训练计划项目 |
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| 摘 要： | 为筛选具有工业应用价值的产蛋白酶新菌种,本研究采用酪素培养基进行新菌株筛选,利用生理生化试验和16S rDNA测序进行菌种鉴定,并结合生化方法测定其蛋白酶的酶活。结果表明,本研究筛选的一株产蛋白酶菌株HSU-2 (GenBank登录号:MN315516)为蜡样芽胞杆菌(Bacillus cereus)。该菌种产蛋白酶的最适pH值和温度分别是7.0和60℃,为一种耐高温的中性蛋白酶。该蛋白酶可耐受5.0%过氧化氢、5.0%十二烷基磺酸钠和5.0%去污剂Triton X-100作用;且5.0%去污剂吐温80可显著提高该蛋白酶的酶活力。此外,Mg2+、Zn2+、K+、Ni2+和Cu2+ 5种金属离子可显著降低该蛋白酶酶活力,其中Zn2+抑制作用最强;而Ca2+可提高该蛋白酶酶活力。本研究为产蛋白酶菌株HSU-2的应用开发和大规模生产奠定了理论基础。
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| 关 键 词： | 菌株 分子鉴定 蛋白酶 酶活力 酶学性质 |
| 收稿时间： | 2020-02-06 |
Molecular Identification and Enzymology Analysis of a High Temperature Resistant Protease-Producing Bacillus cereus Strain |
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| SHE Xinsong,SHAO Jianyang,SUN Chunqiao,FANG Ruyue,CHU Yaqi,JIANG Zhiping,DAI Yuting,BAI Xiaohui.Molecular Identification and Enzymology Analysis of a High Temperature Resistant Protease-Producing Bacillus cereus Strain[J].Acta Agriculturae Nucleatae Sinica,2020,34(8):1698-1704. |
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| Authors: | SHE Xinsong SHAO Jianyang SUN Chunqiao FANG Ruyue CHU Yaqi JIANG Zhiping DAI Yuting BAI Xiaohui |
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| Institution: | 1 College of Life and Environment Science, Huangshan University, Huangshan, Anhui 245041; 2 Forestry bureau of Jixi county, Xuancheng, Anhui 245300 |
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| Abstract: | In order to screen new protease-producing strains with industrial application value, the casein medium was used to screen new strains. Then the physiological and biochemical experiments and 16S rDNA sequencing were used to identify the strains. Likewise, the protease activity was determined by biochemical methods. Our results showed that the protease-producing strain HSU-2 (GenBank accession number: MN315516) was Bacillus cereus. The enzyme activity parameters of the protease from strain HSU-2 showed that the optimal pH and temperature of the protease were 7.0 and 60 ?C, respectively, indicating that it is a high temperature resistant neutral protease. The protease can tolerate 5.0% hydrogen peroxide, 5.0% sodium dodecyl sulfonate, and 5.0% Triton x-100, while 5.0% Tween 80 significantly increases the enzyme activity of the protease. Meanwhile, the effects of metal ions on the activity of protease showed that Mg2+, Zn2+, K+, Ni2+, and Cu2+ significantly reduced the activity of protease, among which Zn2+had the strongest inhibitory effect. Conversely, Ca2+ could increase the activity of protease. Our results laid a theoretical foundation for the industrial application and large-scale production of the protease-producing strain HSU-2. |
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| Keywords: | strain molecular identification protease enzyme activity enzymology properties |
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