抗菌肽LL-1的鉴定和生物学特性研究

doi:10.16431/j.cnki.1671-7236.2025.01.045

Abstract

Abstract: 【Objective】 The aim of this study was to obtain the complete gene sequence of antimicrobial peptide LL-1 and study its biological characteristics and antibacterial effect,in order to lay a foundation for further research and development of antimicrobial peptide drugs. 【Methed】 In this study,an antimicrobial peptide,named LL-1,was discovered through transcriptome sequencing and sequence alignment analysis of Conogethes punctiferalis.The complete gene sequence was obtained by RACE technology,and its corresponding amino acid sequence was derived.After cleavage,the mature peptide sequence was analyzed using multiple sequence alignment and chemically synthesized.Subsequently,the antibacterial effect of antimicrobial peptide LL-1 was evaluated by establishing a model of E.coli infection in chicken meat.Finally,the salt and temperature resistance of antimicrobial peptide LL-1 were analyzed. 【Result】 The results showed that the LL-1 gene had a total length of 474 bp with an ORF length of 192 bp encoding 63 amino acids.Mature antimicrobial peptide LL-1 consisted 39 amino acid residues,and belonged to cecropin family.At the concentration of 50 mg/kg,antimicrobial peptide LL-1 could extremely significantly inhibit E.coli growth in chicken meat (P<0.01).The temperature tolerance analysis showed that the antibacterial effect of antimicrobial peptide LL-1 was not significantly affected by temperature within the range of 4-50 ℃.After being treated at 100 ℃ for 1 h,the antibacterial activity of antimicrobial peptide LL-1 was significantly lower than that of antimicrobial peptide LL-1 treated at 37 ℃ (P<0.05).The salt tolerance analysis showed that the antibacterial effect of antimicrobial peptide LL-1 did not show a significant change after being treated with NaCl solutions of 0-200 mmol/L concentration. 【Conclusion】 Antimicrobial peptide LL-1 had good stability and antibacterial activity,and had the potential to be developed into new drugs.

Key words: antibacterial peptides; chemical synthesis; biological characteristics; antibacterial effect; sequence alignment

CLC Number:

S859.79

ZHOU Lingling, WANG Yuhang, LI Yifei, CHAI Yongle, ZHANG Mingliang, ZHANG Yuanchen, LIAN Kaiqi. Identification and Biological Characteristics of Antibacterial Peptide LL-1[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 491-497.

References

[1] WANG G,LI X,WANG Z.APD3:The antimicrobial peptide database as a tool for research and education[J].Nucleic Acids Research,2016,44(D1):1087-1093.
[2] MWANGI J,KAMAU P M,THUKU R C,et al.Design methods for antimicrobial peptides with improved performance[J].Zoological Research,2023,44(6):1095-1114.
[3] HUANG Y,HUANG J,CHEN Y.Alpha-helical cationic antimicrobial peptides:Relationships of structure and function[J].Protein Cell,2010,1(2):143-152.
[4] YI H Y,CHOWDHURY M,HUANG Y D,et al.Insect antimicrobial peptides and their applications[J].Applied Microbiology and Biotechnology,2014,98(13):5807-5822.
[5] ERDEM BÜYÜKKIRAZ M,KESMEN Z.Antimicrobial peptides (AMPs):A promising class of antimicrobial compounds[J].Journal of Applied Microbiology,2022,132(3):1573-1596.
[6] YANG H,MA R,CHEN J,et al.Discovery of melittin as triple-action agent:Broad-spectrum antibacterial,anti-biofilm,and potential anti-quorum sensing activities[J].Molecules,2024,29(3):558.
[7] PICOLI T,PETER C M,ZANI J L,et al.Melittin and its potential in the destruction and inhibition of the biofilm formation by Staphylococcus aureus,Escherichia coli and Pseudomonas aeruginosa isolated from bovine milk[J].Microbial Pathogenesis,2017,112:57-62.
[8] LEE J K,PARK S C,HAHM K S,et al.A helix-PXXP-helix peptide with antibacterial activity without cytotoxicity against MDRPA-infected mice[J].Biomaterials,2014,35(3):1025-1039.
[9] YEAMAN M R,YOUNT N Y.Mechanisms of antimicrobial peptide action and resistance[J].Pharmacological Reviews,2003,55(1):27-55.
[10] ZHOU J,MCCLEAN S,THOMPSON A,et al.Purification and characterization of novel antimicrobial peptides from the skin secretion of Hylarana guentheri[J].Peptides,2006,27(12):3077-3084.
[11] MA Y,LIU C,LIU X,et al.Peptidomics and genomics analysis of novel antimicrobial peptides from the frog,Rana nigrovittata[J].Genomics,2010,95(1):66-71.
[12] YANG H,LU B,ZHOU D,et al.Identification of the first cathelicidin gene from skin of Chinese giant salamanders Andrias davidianus with its potent antimicrobial activity[J].Developmental and Comparative Immunology,2017,77:141-149.
[13] BARANDOC K P,KIM J,KIM Y.Cotesia plutellae Bracovirus suppresses expression of an antimicrobial peptide,cecropin,in the diamondback moth,Plutella xylostella,challenged by bacteria[J].Journal of Microbiology,2010,48(1):117-123.
[14] WANG H,MENG X L,XU J P,et al.Production,purification,and characterization of the cecropin from Plutella xylostella,pxCECA1,using an intein-induced self-cleavable system in Escherichia coli[J].Applied Microbiology and Biotechnology,2012,94(4):1031-1039.
[15] 刘双双,秦保亮,朱春玲,等.抗菌肽抗病毒作用机制和应用研究进展[J].动物医学进展,2022,43(9):100-104. LIU S S,QIN B L,ZHU C L,et al.Research progress on antiviral mechanism and application of antimicrobial peptides[J].Progress in Veterinary Medicine,2022,43(9):100-104.(in Chinese)
[16] MA Z,YANG J,HAN J,et al.Insights into the antimicrobial activity and cytotoxicity of engineered α-helical peptide amphiphiles[J].Journal of Medicinal Chemistry,2016,59(24):10946-10962.
[17] 李亚菲,张秀凤,张灵璇,等.应对细菌耐药策略研究进展[J].中国畜牧兽医,2023,50(11):4568-4576. LI Y F,ZHANG X F,ZHANG L X,et al.Research progress on strategies to deal with bacterial drug resistance[J].China Animal Husbandry & Veterinary Medicine,2023,50(11):4568-4576.(in Chinese)
[18] 陆鑫,由屹先,吕晓菊.多黏菌素类抗生素治疗碳青霉烯类耐药菌所致重症感染临床研究进展[J].中国感染与化疗杂志,2024,24(2):224-230. LU X,YOU Y X,LYU X J.Progress of clinical research on the treatment of severe infection caused by carbapenem resistant bacteria with polycolistin antibiotics[J].Chinese Journal of Infection and Chemotherapy,2024,24(2):224-230.(in Chinese)
[19] 张丽仕.天然抗菌肽Jelleine-1的结构优化与体内外活性、毒性研究[D].兰州:兰州大学,2021. ZHANG L S.Optimization of structure,activity and toxicity of natural antimicrobial peptide Jelleine-1 in vitro and in vivo[D].Lanzhou:Lanzhou University,2021.(in Chinese)
[20] ZHOU L L,LIAN K Q,WANG M T,et al.The antimicrobial effect of a novel peptide LL-1 on Escherichia coli by increasing membrane permeability[J].BMC Microbiology,2022,22(1):220.
[21] TANG Z,YIN Y,ZHANG Y,et al.Effects of dietary supplementation with an expressed fusion peptide bovine lactoferricin-lactoferrampin on performance,immune function and intestinal mucosal morphology in piglets weaned at age 21 d[J].British Journal of Nutrition,2009,101(7):998-1005.
[22] WU S,ZHANG F,HUANG Z,et al.Effects of the antimicrobial peptide cecropin AD on performance and intestinal health in weaned piglets challenged with Escherichia coli[J].Peptides,2012,35(2):225-230.

Identification and Biological Characteristics of Antibacterial Peptide LL-1

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