| 嗜水气单胞菌群体感应信号分子AI-2的细胞外生物
合成及活性检测 |
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| 引用本文： | 赵晶,张福蓉,崔一,许永斌,陈明,权春善,范圣第.嗜水气单胞菌群体感应信号分子AI-2的细胞外生物
合成及活性检测[J].微生物学通报,2015,42(10):1858-1865. |
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| 作者姓名： | 赵晶 张福蓉 崔一 许永斌 陈明 权春善 范圣第 |
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| 作者单位： | 1. 大连民族大学生命科学学院 国家民委-教育部生物化工重点实验室 辽宁 大连 116600,1. 大连民族大学生命科学学院 国家民委-教育部生物化工重点实验室 辽宁 大连 116600,2. 大连工业大学 生物工程学院 辽宁 大连 116034,1. 大连民族大学生命科学学院 国家民委-教育部生物化工重点实验室 辽宁 大连 116600,2. 大连工业大学 生物工程学院 辽宁 大连 116034,1. 大连民族大学生命科学学院 国家民委-教育部生物化工重点实验室 辽宁 大连 116600,1. 大连民族大学生命科学学院 国家民委-教育部生物化工重点实验室 辽宁 大连 116600 |
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| 基金项目： | 国家自然科学基金项目(No. 21272031)；辽宁省教育厅一般项目(No. L2013508) |
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| 摘 要： | 【目的】对嗜水气单胞菌群体感应信号分子AI-2进行细胞外生物合成及活性检测。【方法】对LuxS、MtnN-1、MtnN-2蛋白进行氨基酸序列分析、表达及纯化。以S-腺苷同型半胱氨酸(SAH)为底物，利用纯化的LuxS分别与MtnN-1及MtnN-2蛋白共同作用合成AI-2，并利用哈维氏弧菌报告菌株BB170检测AI-2活性。【结果】嗜水气单胞菌培养液上清中AI-2活性在8 h达到空白对照的16.96倍。氨基酸序列分析表明，嗜水气单胞菌与水生病原菌哈维式弧菌和迟钝爱德华氏LuxS一致性达到76%以上，MtnN-1与MtnN-2氨基酸序列一致性为26.37%，其中MtnN-2与哈维氏弧菌和迟钝爱德华氏菌Pfs一致性达到53%以上。成功表达及纯化了LuxS、MtnN-1和MtnN-2蛋白，细胞外LuxS和MtnN-1共同作用合成的AI-2活性是空白对照的45.04倍，LuxS和MtnN-2共同作用合成的AI-2活性是空白对照的63.62倍。【结论】嗜水气单胞菌能够合成信号分子AI-2。MtnN-1和MtnN-2氨基酸序列尽管存在较大差异，但两者均能与LuxS共同催化AI-2的细胞外生物合成。
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| 关 键 词： | 嗜水气单胞菌，群体感应信号分子AI-2，生物合成，LuxS，MtnN |
In vitro biosynthesis and activity detection of quorum sensing
signal molecule AI-2 of Aeromonas hydrophila |
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| ZHAO Jing,ZHANG Fu-Rong,CUI Yi,XU Yong-Bin,CHEN Ming,QUAN Chun-Shan and FAN Sheng-Di.In vitro biosynthesis and activity detection of quorum sensing
signal molecule AI-2 of Aeromonas hydrophila[J].Microbiology,2015,42(10):1858-1865. |
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| Authors: | ZHAO Jing ZHANG Fu-Rong CUI Yi XU Yong-Bin CHEN Ming QUAN Chun-Shan and FAN Sheng-Di |
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| Institution: | 1. College of Life Science, Dalian Nationalities University, State Ethnic Affairs Commission-Ministry of Education Key Laboratory of Biochemical Engineering, Dalian Nationalities University, Dalian, Liaoning 116600, China,1. College of Life Science, Dalian Nationalities University, State Ethnic Affairs Commission-Ministry of Education Key Laboratory of Biochemical Engineering, Dalian Nationalities University, Dalian, Liaoning 116600, China,2. School of Biological Engineering, Dalian Polytechnic University, Dalian, Liaoning 116034, China,1. College of Life Science, Dalian Nationalities University, State Ethnic Affairs Commission-Ministry of Education Key Laboratory of Biochemical Engineering, Dalian Nationalities University, Dalian, Liaoning 116600, China,2. School of Biological Engineering, Dalian Polytechnic University, Dalian, Liaoning 116034, China,1. College of Life Science, Dalian Nationalities University, State Ethnic Affairs Commission-Ministry of Education Key Laboratory of Biochemical Engineering, Dalian Nationalities University, Dalian, Liaoning 116600, China and 1. College of Life Science, Dalian Nationalities University, State Ethnic Affairs Commission-Ministry of Education Key Laboratory of Biochemical Engineering, Dalian Nationalities University, Dalian, Liaoning 116600, China |
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| Abstract: | Objective] To realize in vitro biosynthesis and activity detection of quorum sensing signal molecule autoinducer 2 (AI-2) of Aeromonas hydrophila. Methods] LuxS, MtnN-1, MtnN-2 proteins were analyzed for amino acid sequences, expressed and purified. AI-2 was synthesized from S-adenosylhomocysteine (SAH) with purified LuxS and MtnN-1, or purified LuxS and MtnN-2 separately. The activity of AI-2 was detected by the reporter strain Vibrio harveyi BB170. Results] The activity of AI-2 in A. hydrophila culture supernatant was 16.96-fold of the control at 8 h. As analysis of amino acid sequence alignment indicated, the identity of LuxS of A. hydrophila with that of V. harveyi and Edwardsiella tarda is above 76%. The identity of MtnN-1 with MtnN-2 is 26.37%. MtnN-2 has relatively high homologies with representative Pfs amino acid sequences. The identity of MtnN-2 with that of V. harveyi and E. tarda is above 53%. LuxS, MtnN-1 and MtnN-2 proteins were successfully expressed and purified. The activity of AI-2 produced with purified LuxS and MtnN-1 in vitro was 45.04-fold of the control; the activity of AI-2 produced with purified LuxS and MtnN-2 in vitro was 63.62-fold of the control. Conclusion] A. hydrophila could secrete AI-2. MtnN-1 and MtnN-2 could both cooperate with LuxS and realize in vitro biosynthesis of AI-2, though the amino acid sequences between the two showed relatively big differences. |
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| Keywords: | Aeromonas hydrophila Quorum sensing signal molecule AI-2 Biosynthesis LuxS MtnN |
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