Vibrio alginolyticus is a zoonotic bacterium.A pair of specific primers was designed using the sodB gene sequence of Vibrio alginolyticus HY9901 in order to amplify the full length of the gene by PCR.The results indic...Vibrio alginolyticus is a zoonotic bacterium.A pair of specific primers was designed using the sodB gene sequence of Vibrio alginolyticus HY9901 in order to amplify the full length of the gene by PCR.The results indicated that the total length of the sodB gene was 585 bp and that it could encode 194 amino acids.The predicted amino acid sequence derivation indicated that the molecular weight of the protein was approximately 21.56 kDa,with an isoelectric point of 4.95.Upon prediction of the N-terminal signal peptide structure of the protein,no significant signal peptide cleavage site was observed,indicating that the protein lacked both a signal peptide and a transmembrane region.The amino acid sequence contained an N-glycosylation site,a casein kinase II phosphorylation site,a microsomal C-terminal target signal site,and a manganese and iron superoxide dismutase signal site.The probability of intracytoplasmic localization of the SodB protein was 56.5%,which was analyzed according to the subcellular localization of the protein.The amino acid sequence of the sodB gene of V.alginolyticus exhibited 98%-100%homology to other Vibrio species,clustering into the same subfamily with V.parahaem,indicating a relatively close relationship between them.In the prediction of protein structure,the proportions ofα-helix,random coil,β-sheet,and extended strand were 48.45%,30.41%,5.67%,and 15.46%,respectively.The similarity to template 1dt0.1.A reached 71.58%.A PTM site analysis revealed the presence of phosphorylation,glycosylation,ubiquitination,sumoylation,acetylation,and methylation modification sites,as well as the absence of lactylation modification sites.展开更多
BACKGROUND Resistance to antibiotics is one the main factors constraining the treatment and control of Helicobacter pylori(H.pylori)infections.Therefore,there is an urgent need to develop new antimicrobial agents to r...BACKGROUND Resistance to antibiotics is one the main factors constraining the treatment and control of Helicobacter pylori(H.pylori)infections.Therefore,there is an urgent need to develop new antimicrobial agents to replace antibiotics.Our previous study found that linolenic acid-metronidazole(Lla-Met)has a good antibacterial effect against H.pylori,both antibiotic-resistant and sensitive H.pylori.Also,H.pylori does not develop resistance to Lla-Met.Therefore,it could be used for preparing broad-spectrum antibacterial agents.However,since the antibacterial mechanism of Lla-Met is not well understood,we explored this phenomenon in the present study.AIM To understand the antimicrobial effect of Lla-Met and how this could be applied in treating corresponding infections.METHODS H.pylori cells were treated with the Lla-Met compound,and the effect of the compound on the cell morphology,cell membrane permeability,and oxidation of the bacteria cell was assessed.Meanwhile,the differently expressed genes in H.pylori in response to Lla-Met treatment were identified.RESULTS Lla-Met treatment induced several changes in H.pylori cells,including roughening and swelling.In vivo experiments revealed that Lla-Met induced oxidation,DNA fragmentation,and phosphatidylserine ectropionation in H.pylori cells.Inhibiting Lla-Met with L-cysteine abrogated the above phenomena.Transcriptome analysis revealed that Lla-Met treatment up-regulated the expression of superoxide dismutase SodB and MdaB genes,both anti-oxidation-related genes.CONCLUSION Lla-Met kills H.pylori mainly by inducing oxidative stress,DNA damage,phosphatidylserine ectropionation,and changes on cell morphology.展开更多
基金Supported by Outstanding Graduate Entering Laboratory Project of College of Fisheries,Guangdong Ocean UniversityNational Natural Science Foundation of China(32073015)+1 种基金Undergraduate Innovation and Entrepreneurship Training Program of Guangdong Ocean University(CXXL2024007)Undergraduate Innovation Team of Guangdong Ocean University(CCTD201802).
文摘Vibrio alginolyticus is a zoonotic bacterium.A pair of specific primers was designed using the sodB gene sequence of Vibrio alginolyticus HY9901 in order to amplify the full length of the gene by PCR.The results indicated that the total length of the sodB gene was 585 bp and that it could encode 194 amino acids.The predicted amino acid sequence derivation indicated that the molecular weight of the protein was approximately 21.56 kDa,with an isoelectric point of 4.95.Upon prediction of the N-terminal signal peptide structure of the protein,no significant signal peptide cleavage site was observed,indicating that the protein lacked both a signal peptide and a transmembrane region.The amino acid sequence contained an N-glycosylation site,a casein kinase II phosphorylation site,a microsomal C-terminal target signal site,and a manganese and iron superoxide dismutase signal site.The probability of intracytoplasmic localization of the SodB protein was 56.5%,which was analyzed according to the subcellular localization of the protein.The amino acid sequence of the sodB gene of V.alginolyticus exhibited 98%-100%homology to other Vibrio species,clustering into the same subfamily with V.parahaem,indicating a relatively close relationship between them.In the prediction of protein structure,the proportions ofα-helix,random coil,β-sheet,and extended strand were 48.45%,30.41%,5.67%,and 15.46%,respectively.The similarity to template 1dt0.1.A reached 71.58%.A PTM site analysis revealed the presence of phosphorylation,glycosylation,ubiquitination,sumoylation,acetylation,and methylation modification sites,as well as the absence of lactylation modification sites.
文摘BACKGROUND Resistance to antibiotics is one the main factors constraining the treatment and control of Helicobacter pylori(H.pylori)infections.Therefore,there is an urgent need to develop new antimicrobial agents to replace antibiotics.Our previous study found that linolenic acid-metronidazole(Lla-Met)has a good antibacterial effect against H.pylori,both antibiotic-resistant and sensitive H.pylori.Also,H.pylori does not develop resistance to Lla-Met.Therefore,it could be used for preparing broad-spectrum antibacterial agents.However,since the antibacterial mechanism of Lla-Met is not well understood,we explored this phenomenon in the present study.AIM To understand the antimicrobial effect of Lla-Met and how this could be applied in treating corresponding infections.METHODS H.pylori cells were treated with the Lla-Met compound,and the effect of the compound on the cell morphology,cell membrane permeability,and oxidation of the bacteria cell was assessed.Meanwhile,the differently expressed genes in H.pylori in response to Lla-Met treatment were identified.RESULTS Lla-Met treatment induced several changes in H.pylori cells,including roughening and swelling.In vivo experiments revealed that Lla-Met induced oxidation,DNA fragmentation,and phosphatidylserine ectropionation in H.pylori cells.Inhibiting Lla-Met with L-cysteine abrogated the above phenomena.Transcriptome analysis revealed that Lla-Met treatment up-regulated the expression of superoxide dismutase SodB and MdaB genes,both anti-oxidation-related genes.CONCLUSION Lla-Met kills H.pylori mainly by inducing oxidative stress,DNA damage,phosphatidylserine ectropionation,and changes on cell morphology.
