[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin...[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin B was synthesized by SOE-PCR with six primers designed according to the published amino acid sequences. Then, the BC gene fragment was ligated into pET32a vector and expressed in BI21. Antimierobial ability of the crude hybrid antimicrobial peptide was detected with the Oxford cup method. [ Result] The BC gene fragment was correctly amplified by SOE-PCR and ligated into pET32a vector, to construct the expression vector pET32-BC. The hybrid antimicrobial peptide buforin II-cecropin B was expressed in BI21 after induction by IPTG. The optimal induction time was 2 h. Antimicrobial activity assay suggested that the hybrid antimicrobial peptide possessed antimicrobial activity to Escherichia coli and Bacillus subtilis. [ Conclusion] The hybrid antimicrobial peptide buforin II-cecropin B exhibited antimicrobial activity to Gram-negative E. coli and Gram-positive B. subtilis strains.展开更多
In the last decade,the rise of antibiotic resistance has heightened interest in antimicrobial peptides and lipopeptides as promising alternatives to conventional antibiotics because of their lower propensity to develo...In the last decade,the rise of antibiotic resistance has heightened interest in antimicrobial peptides and lipopeptides as promising alternatives to conventional antibiotics because of their lower propensity to develop resistance.However,lipopeptides often show undesired cytotoxicity due to their non-selective membrane disruptive effect,and their limited aqueous solubility represents a matter of concern from a pharmaceutical point of view.This study demonstrates a panel of ultrashort cationic lipopeptides(USCLs)consisting of a tetrapeptide(L1),originated from buforin II,coupled with saturated fatty acids of different lengths.Our results highlight that the 16-carbon fatty acid lipopeptide(Pal-L1)exhibits relevant antibacterial activity against multiresistant Staphylococcus aureus strain.However,the formation of heterogenic aggregates in cell culturemedium and toxic effects on human cells were also observed.Pal-L1 formulation with the randomly methylatedα-cyclodextrin(RAMEA)and the sulfobutylether-β-cyclodextrin(SBECD)has resulted in a production of ultralow-sized molecular dispersion systems and reduced lipopeptide toxicity without compromising its antimicrobial activity.With titration 1H-NMR,2D NMR experiments,together with molecular dynamics simulations,we described the size,structure,stoichiometry,and dissociation constant of the supramolecular complexes.Interactions of neutral and negatively chargedmodel liposomes with Pal-L1 lipopeptide in the presence or absence of cyclodextrins serve an explanation for the membrane selectivity,and based on the results,we proposed a potential mechanism of action for the Pal-L1+cyclodextrin complexes on different biological membranes.Overall,our model characterization points out that cyclodextrin formulation improves the therapeutical applicability of lipopeptides.展开更多
基金Supported by the Research Project of Sichuan University of Science&Engineering(No.2011RC12,2014KY02)Scientific Research Foundation of the Education Department of Sichuan Province(NO.15ZA0222)
文摘[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin B was synthesized by SOE-PCR with six primers designed according to the published amino acid sequences. Then, the BC gene fragment was ligated into pET32a vector and expressed in BI21. Antimierobial ability of the crude hybrid antimicrobial peptide was detected with the Oxford cup method. [ Result] The BC gene fragment was correctly amplified by SOE-PCR and ligated into pET32a vector, to construct the expression vector pET32-BC. The hybrid antimicrobial peptide buforin II-cecropin B was expressed in BI21 after induction by IPTG. The optimal induction time was 2 h. Antimicrobial activity assay suggested that the hybrid antimicrobial peptide possessed antimicrobial activity to Escherichia coli and Bacillus subtilis. [ Conclusion] The hybrid antimicrobial peptide buforin II-cecropin B exhibited antimicrobial activity to Gram-negative E. coli and Gram-positive B. subtilis strains.
基金The authors acknowledge the support of the Lendulet(Momentum)Programme of the Hungarian Academy of Sciences(Grant No.:LP2021-28)National Research,Development and Innovation Fund of Hungary(Grant Nos.:OTKA K142904,OTKA ANN 139484,KKP_22144180,TKP2021-EGA-31,and VEKOP-2.3.3-15-2016-00020)+2 种基金Marie-Skłodowska-Curie action(H2020-MSCA-ITN BactiVAX,grant agreement No.956758)CIBERES,an initiative of Instituto de Salud Carlos III(ISCIII,Madrid,Spain)Agencia Estatal Investigacion of Spain for the Severo Ochoa Center of Excellence Accreditation CEX2021-001136-S,funded by MCIN/AEI/10.13039/501100011033.
文摘In the last decade,the rise of antibiotic resistance has heightened interest in antimicrobial peptides and lipopeptides as promising alternatives to conventional antibiotics because of their lower propensity to develop resistance.However,lipopeptides often show undesired cytotoxicity due to their non-selective membrane disruptive effect,and their limited aqueous solubility represents a matter of concern from a pharmaceutical point of view.This study demonstrates a panel of ultrashort cationic lipopeptides(USCLs)consisting of a tetrapeptide(L1),originated from buforin II,coupled with saturated fatty acids of different lengths.Our results highlight that the 16-carbon fatty acid lipopeptide(Pal-L1)exhibits relevant antibacterial activity against multiresistant Staphylococcus aureus strain.However,the formation of heterogenic aggregates in cell culturemedium and toxic effects on human cells were also observed.Pal-L1 formulation with the randomly methylatedα-cyclodextrin(RAMEA)and the sulfobutylether-β-cyclodextrin(SBECD)has resulted in a production of ultralow-sized molecular dispersion systems and reduced lipopeptide toxicity without compromising its antimicrobial activity.With titration 1H-NMR,2D NMR experiments,together with molecular dynamics simulations,we described the size,structure,stoichiometry,and dissociation constant of the supramolecular complexes.Interactions of neutral and negatively chargedmodel liposomes with Pal-L1 lipopeptide in the presence or absence of cyclodextrins serve an explanation for the membrane selectivity,and based on the results,we proposed a potential mechanism of action for the Pal-L1+cyclodextrin complexes on different biological membranes.Overall,our model characterization points out that cyclodextrin formulation improves the therapeutical applicability of lipopeptides.
