新型黏虫抗菌肽AAP-1对大肠杆菌的抗菌作用

doi:10.16431/j.cnki.1671-7236.2020.11.025

Abstract

Abstract: In order to evaluate the bactericidal activity of a novel armyworm antimicrobial peptide (AAP-1),the bactericidal effect of AAP-1 on Escherichia coli (E.coli) was investigated.Firstly,AAP-1 was synthesized by solid-phase chemically synthesis,and purified by high-performance liquid chromatography and tested by mass spectrometry.Then,the bactericidal effect of AAP-1 against E.coli was determined by the bacteriostatic circle method.The minimum inhibitory concentration (MIC) was detected by the double dilution method.The time kill curve was measured by bacterial plate counting method.Finally,the effect of AAP-1 on the nucleic acid leakage of E.coli was evaluation by ultra-micro spectrophotometer.The effect of AAP-1 on the intracellular DNA of E.coli was tested by nucleic acid gel electrophoresis.The overall effect of AAP-1 on E.coli was observed through transmission electron microscope.The results showed that the purity of the synthesized AAP-1 was more than 98%,and its molecular weight was 4 262.17 u.AAP-1 showed good antibacterial activity against E.coli,with a MIC of 7.8 μg/mL.AAP-1 had a concentration-dependent bactericidal effect on E.coli,and within 60 min the bactericidal effect gradually increased.AAP-1 could cause the nucleic acid leakage of E.coli,and resulted in a reduction in the total amount of intracellular DNA of E.coli with concentration-dependent.AAP-1 could also cause cell membrane destruction,significantly reduce intracellular electron density,resulted in cytoplasmic dissolution of E.coli,and so on.In summary,this study suggested that a novel antimicrobial peptide AAP-1 could be chemically synthesized with a purity of up to 98% and showed its good bactericidal effect on E.coli.This study could provide preliminary data for the in-depth study of the bactericidal mechanism of AAP-1 against E.coli and laid a theoretical foundation for the clinical application of AAP-1.

Key words: antimicrobial peptide; Escherichia coli; antibacterial curve; cell membrane

CLC Number:

S852.61

LIAN Kaiqi, ZHOU Lingling, ZHANG Yuanchen, SHI Zhiqi, LIN Zhengdan, SONG Yuwei, WANG Shuangshan, ZHANG Mingliang. Antibacterial Effect of Novel Armyworm Antimicrobial Peptide (AAP-1) Against Escherichia coli[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3626-3632.

References

[1] BROWN E D,WRIGHT G D.Antibacterial drug disco-very in the resistance era[J].Nature,2016,529(7586):336-343.
[2] RAO L L,LV L C,ZENG Z L,et al.Increasing prevalence of extended-spectrum cephalosporin-resistant Escherichia coli in food animals and the diversity of CTX-M genotypes during 2003-2012[J].Veterinary Microbiology,2014,172(3-4):534-541.
[3] TIBERI S,ZUMLA A,MIGLIORI G B.Multidrug and extensively drug-resistant tuberculosis:Epidemiology,clinical features,management and treatment[J].Infectious Disease Clinics of North America,2019,33(4):1063-1085.
[4] DA SILVA A C,RODRIGUES M X,SILVA N C C.Methicillin-resistant Staphylococcus aureus in food and the prevalence in Brazil:A review[J].Brazilian Journal of Microbiology,2020,51(1):347-356.
[5] VAN BOECKEL T P,PIRES J,SILVESTER R,et al.Global trends in antimicrobial resistance in animals in low- and middle-income countries[J].Science,2019,365(6459):eaaw1944.
[6] BAKKEREN E,HUISMAN J S,FATTINGER S A,et al.Salmonella persisters promote the spread of antibiotic resistance plasmids in the gut[J].Nature,2019,573(7773):276-280.
[7] PIRES J,SIRIWARDENA T N,STACH M,et al. In vitro activity of the novel antimicrobial peptide dendrimer G3KL against multidrug-resistant acinetobac-terbaumannii and Pseudomonas aeruginosa[J].Antimi-crobial Agents and Chemotherapy,2015,59(12):7915-7918.
[8] VERGIS J,MALIK S V S,PATHAK R,et al.Efficacy of indolicidin,cecropin A (1-7)-melittin (CAMA) and their combination against biofilm-forming multidrug-resistant enteroaggregative Escherichia coli[J].Probiotics and Antimicrobial Proteins 2020,12(2):705-715.
[9] ALMATAR M,MAKKY E A,YAKICI G,et al.Antimicrobial peptides as an alternative to anti-tuberculosis drugs[J].Pharmacological Research,2018,128:288-305.
[10] ZHENG Z,THARMALINGAM N,LIU Q,et al.Synergistic efficacy of Aedesaegyptiantimicrobial peptide cecropin A2 and tetracycline against Pseudomonas aeruginosa[J].Antimicrobial Agents and Chemotherapy,2017,61(7):e00686-17.
[11] LIAN K Q,ZHANG M L,LIANG X L,et al.Identification and characteristics of a novel cecropin from the armyworm,Mythimna separata[J].BMC Microbiology,2020,20(1):233.
[12] SHEN Z Q,WANG Y,SHEN Y B,et al.Early emergence of mcr-1 in Escherichia coli from food-producing animals[J].The Lancet Infectious Diseases,2016,16(3):293.
[13] PEI Z H,BIAN L,ZHANG H,et al.Cloning,expression,and purification of a new antibacterial substance gene from Larvae of Muscadomestica (Diptera:Muscidae)[J].Journal of Insect Science,2014,14:253.
[14] WANG L,ZHAO X Q,XIA X J,et al.Antimicrobial peptide JH-3 effectively kills Salmonella enterica serovar Typhimurium strain CVCC541 and reduces its pathogenicity in mice[J].Probiotics and Antimi-crobial Proteins,2019,11(4):1379-1390.
[15] 钟亨任,谢志鹏,魏涵淇,等.抗菌肽Brevinin-GR23抗金黄色葡萄球菌作用机制的初步研究[J].海南大学学报(自然科学版),2018,36(4):360-367. ZHONG H R,XIE Z P,WEI H Q,et al.Mechanism of Brevinin-GR23 against Staphylococcus aureus[J].Natural Science Journal of Hainan University,2018,36(4):360-367.(in Chinese)
[16] 王青,徐彦召,杨磊,等.一种广谱、低毒的抗菌肽类似物HJH-3的生物学活性和膜活性的测定[J].中国预防兽医学报,2019,41(8):800-806. WANG Q,XU Y Z,YANG L,et al.Determination of bioactivity and membrane activity of bovine antimicrobial peptide HJH-3[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(8):800-806.(in Chinese)
[17] HILCHIE A L,WUERTH K,HANCOCK R E.Immune modulation by multifaceted cationic host defense (antimicrobial) peptides[J].Nature Chemical Biology,2013,9(12):761-768.
[18] LAI Y P,GALLO R L.AMPed up immunity:How antimicrobial peptides have multiple roles in immune defense[J].Trends in Immunology,2009,30(3):131-141.
[19] MWANGI J,HAO X,LAI R,et al.Antimicrobial peptides:New hope in the war against multidrug resistance[J].Zoological Research,2019,40(6):488-505.
[20] GUANI-GUERRA E,SANTOS-MENDOZA T,LUGO-REYES S O,et al.Antimicrobial peptides:General overview and clinical implications in human health and disease[J].Clinical Immunology,2010,135(1):1-11.
[21] ZHANG Q H,XU Y Z,WANG Q,et al.Potential of novel antimicrobial peptide P3 from bovine erythrocytes and its analogs to disrupt bacterial membranes in vitro and display activity against drug-resistant bacteria in a mouse model[J].Antimicrobial Agents and Chemotherapy,2015,59(5):2835-2841.
[22] WANG X M,TENG D,MAO R Y,et al.Combined systems approaches reveal a multistage mode of action of a marine antimicrobial peptide against pathogenic Escherichia coli and its protective effect against bacterial peritonitis and endotoxemia[J].Antimi-crobial Agents and Chemotherapy,2017,61(1):e01056-16.
[23] DONG Z,LUO W J,ZHONG H R,et al.Molecular cloning and characterization of antimicrobial peptides from skin of Hylaranaguentheri[J].Acta Biochimica et Biophysica Sinica,2017,49(5):450-457.
[24] LIU Y,SONG M R,DING S Y,et al.Discovery of linear low-cationic peptides to target methicillin-resistant Staphylococcus aureus in vivo[J].ACS Infectious Diseases,2019,5(1):123-130.

Antibacterial Effect of Novel Armyworm Antimicrobial Peptide (AAP-1) Against Escherichia coli

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