抗菌肽Tachyplesin Ⅰ的分子设计、结构与活性分析

doi:10.11843/j.isn.0366-6964.2022.06.024

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (6): 1905-1913.doi: 10.11843/j.isn.0366-6964.2022.06.024

抗菌肽Tachyplesin Ⅰ的分子设计、结构与活性分析

孙栋¹, 蔡寅川², 蒋思雨¹, 李璇³, 郝刚^1*

1. 西南民族大学食品科学与技术学院，成都 610041;
2. 四川阿坝州工业经济研究所，汶川 623000;
3. 重庆市计量质量检测研究院，重庆 401120

收稿日期:2021-06-20 出版日期:2022-06-23 发布日期:2022-06-25
通讯作者: 郝刚，主要从事多肽的生物活性及构效关系研究，E-mail: indianahg@hotmail.com，Tel: 86-28-85928240
作者简介:孙栋(1994-)，男，甘肃庆阳人，硕士生，主要从事畜产品加工与安全研究，E-mail: 441506324@qq.com
基金资助:
国家重点研发计划(2018YFD0400101)；西南民族大学研究生创新型科研项目(CX2021SZ75)

Molecular Design, Structure and Activity Analysis of Antimicrobial Peptide Tachyplesin I

SUN Dong¹, CAI Yinchuan², JIANG Siyu¹, LI Xuan³, HAO Gang^1*

1. College of Food Science and Technology, Southwest Minzu University, Chengdu 610041, China;
2. The Research Institute of Industrial Economy of Aba, Wenchuan 623000, China;
3. Chongqing Academy of Metrology and Quality Inspection, Chongqing 401120, China

Received:2021-06-20 Online:2022-06-23 Published:2022-06-25

摘要/Abstract

摘要： 本研究旨在抗菌肽Tachyplesin Ⅰ(TP Ⅰ)的结构活性基础上，重新设计一种全新的抗菌肽TP Ⅰ-Y4。保持母肽链长与电荷数不变，将序列中形成TP Ⅰ两个二硫键的4个半胱氨酸用酪氨酸进行替换后得到TP Ⅰ-Y4。经生物信息学软件分析，与TP Ⅰ相比，TP Ⅰ-Y4的热稳定性更好，亲水性更强，同时具有与母肽相似的结构与抗菌活性。化学合成后经圆二色谱检测发现，TP Ⅰ-Y4在水相及在模拟细胞膜疏水环境的50%三氟乙醇中都表现出β-折叠结构，疏水环境中TP Ⅰ-Y4的β-折叠含量高于水相，也高于不同环境中的TP Ⅰ。抑菌试验表明，TP Ⅰ-Y4对细菌和真菌都有较强抑菌活性，且对细菌的抑菌活性强于TP Ⅰ。TP Ⅰ-Y4对小鼠红细胞溶血性降低，并保留了很强的内毒素中和能力，浓度为40 μg·mL^-1时中和率可达50%以上。TP Ⅰ-Y4抗菌活性的提高以及溶血活性的降低，可能与β-折叠强形成者酪氨酸的替代有关，进而导致抗菌肽分子中β-折叠结构增强。

关键词: 抗菌肽, Tachyplesin Ⅰ, 分子设计, 圆二色谱, 抑菌活性

Abstract: Based on the structure-activity relationship of the antimicrobial peptide tachyplesin Ⅰ(TP Ⅰ), a novel antimicrobial peptide TP Ⅰ-Y4 was designed out. TP Ⅰ-Y4 was obtained by replacing four cysteines with tyrosines to delete two disulfide bonds of TP Ⅰ, keeping the chain length and charge number of the parent peptide. The results of bioinformatics software analysis had discovered that compared with TP Ⅰ, TP Ⅰ-Y4 had better thermal stability, stronger hydrophilicity, and similar structure and antibacterial activity to propeptide. After chemical synthesis, the results of circular dichroism spectroscopy had showed that TP Ⅰ-Y4 also adopted β-sheet structure in aqueous phase and 50% trifluoroethanol solution which simulated the hydrophobic environment of cell membrane. The β-sheet contents of TP Ⅰ-Y4 in hydrophobic environment were higher than that in aqueous phase. TP Ⅰ-Y4 exhibited higher β-sheet contents than TP Ⅰ in different environments. TP Ⅰ-Y4 owned potent antimicrobial activities against bacteria and fungi, and TP Ⅰ-Y4 displayed stronger antimicrobial activities against bacteria tested than TP Ⅰ. TP Ⅰ-Y4 reduced the hemolysis of mouse red blood cells and retained a strong endotoxin neutralization ability, the neutralization rate reached more than 50% when the concentration was 40 μg·mL^-1. The results suggest that the increase of TP I-Y4 antibacterial activity and the decrease of hemolytic activity may be related to the substitution of tyrosine, as β-sheet strong-former which further leads to the enhancement of β-sheet structure in antimicrobial peptide.

Key words: antimicrobial peptide, Tachyplesin Ⅰ, molecular design, circular dichroism, antimicrobial activity

中图分类号:

S816

孙栋, 蔡寅川, 蒋思雨, 李璇, 郝刚. 抗菌肽Tachyplesin Ⅰ的分子设计、结构与活性分析[J]. 畜牧兽医学报, 2022, 53(6): 1905-1913.

SUN Dong, CAI Yinchuan, JIANG Siyu, LI Xuan, HAO Gang. Molecular Design, Structure and Activity Analysis of Antimicrobial Peptide Tachyplesin I[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1905-1913.

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抗菌肽Tachyplesin Ⅰ的分子设计、结构与活性分析

Molecular Design, Structure and Activity Analysis of Antimicrobial Peptide Tachyplesin I

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