一种猪溶菌酶来源的抗菌六肽的分离鉴定及其性质
发布时间：2018-12-18 18:03
【摘要】：为提高猪溶菌酶(Sus scrofa lysozyme,SSL)的抗革兰氏阴性菌活性,将其进行了不同蛋白酶的水解,选择抗革兰氏阴性菌效果最好的水解产物,利用凝胶过滤色谱和反相制备色谱进行分离,对其功能成分进行液质联用鉴定。对分离得到的物质进行抗菌活性验证和生物信息学的分析,并在此基础上对抗菌物质的杀菌机理进行了探讨。结果表明,胰蛋白酶的水解产物具有较高的杀灭革兰氏阴性菌的活性,进一步分离纯化得到了具有抗革兰氏阴性菌活性的六肽A-W-V-A-W-K。经化学合成验证,该六肽既保留了SSL的部分抗菌活性,也具备杀灭多种革兰氏阴性菌的能力。进一步分析发现其位于SSL分子C端的一个螺旋-回环-螺旋的结构中,并由此推测其杀菌机理是通过改变细胞膜的渗透性,进而使细胞内溶物流出而造成细胞死亡,而抗菌实验也验证了这一推测。该抗菌肽的发现为后续提高SSL的抗菌活性提供了理论依据。
[Abstract]:In order to improve the activity of porcine lysozyme (Sus scrofa lysozyme,SSL against Gram-negative bacteria, different proteases were hydrolyzed. The functional components were separated by gel filtration chromatography and reversed phase preparation chromatography. The antimicrobial activity and bioinformatics of the separated substances were verified and the bactericidal mechanism of the antibacterial substances was discussed. The results showed that the hydrolysate of trypsin had higher activity to kill Gram-negative bacteria, and the six peptide A-W-V-A-W-Kwith the activity of anti-Gram-negative bacteria was obtained by further isolation and purification. It was proved by chemical synthesis that the six peptides not only retained some of the antibacterial activity of SSL, but also had the ability to kill many Gram-negative bacteria. Further analysis revealed that it was located in a helix-loop-helix structure at the C-terminal of SSL molecule, and its bactericidal mechanism was presumed to result in cell death by changing the permeability of the cell membrane and resulting in the release of intracellular solutes. Antimicrobial experiments also confirmed the hypothesis. The discovery of the antimicrobial peptide provides a theoretical basis for improving the antibacterial activity of SSL.
【作者单位】： 江南大学工业生物技术教育部重点实验室;江南大学粮食发酵工艺与技术国家工程实验室;江南大学生物工程学院;
【基金】：国家重点基础研究发展计划(973计划)(No.2013CB733602) 中央高校基本科研业务费专项资金资助(No.JUSRP51302A) 江苏高校优势学科建设工程项目资助~~
【分类号】：Q51
，
本文编号：2386266
[Abstract]:In order to improve the activity of porcine lysozyme (Sus scrofa lysozyme,SSL against Gram-negative bacteria, different proteases were hydrolyzed. The functional components were separated by gel filtration chromatography and reversed phase preparation chromatography. The antimicrobial activity and bioinformatics of the separated substances were verified and the bactericidal mechanism of the antibacterial substances was discussed. The results showed that the hydrolysate of trypsin had higher activity to kill Gram-negative bacteria, and the six peptide A-W-V-A-W-Kwith the activity of anti-Gram-negative bacteria was obtained by further isolation and purification. It was proved by chemical synthesis that the six peptides not only retained some of the antibacterial activity of SSL, but also had the ability to kill many Gram-negative bacteria. Further analysis revealed that it was located in a helix-loop-helix structure at the C-terminal of SSL molecule, and its bactericidal mechanism was presumed to result in cell death by changing the permeability of the cell membrane and resulting in the release of intracellular solutes. Antimicrobial experiments also confirmed the hypothesis. The discovery of the antimicrobial peptide provides a theoretical basis for improving the antibacterial activity of SSL.
【作者单位】： 江南大学工业生物技术教育部重点实验室;江南大学粮食发酵工艺与技术国家工程实验室;江南大学生物工程学院;
【基金】：国家重点基础研究发展计划(973计划)(No.2013CB733602) 中央高校基本科研业务费专项资金资助(No.JUSRP51302A) 江苏高校优势学科建设工程项目资助~~
【分类号】：Q51
，
本文编号：2386266
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