Dataset Information

Effects of 1,8-cineole on Carbohydrate Metabolism Related Cell Structure Changes of Salmonella.

ABSTRACT: Salmonella is gram-negative foodborne zoonotic bacteria which triggers disease in humans. Our previous studies showed 1,8-cineole possessed remarkable antimicrobial effects on foodborne zoonotic bacteria indicating it could serve as a new source of antibiotic for Salmonella. Present study elucidated the antibacterial mechanism of 1,8-cineole by analyzing serum protein expressed by Salmonella following treatment with 1,8-cineole (0.25 mg/mL, 3 h) using isobaric tags for relative and absolute quantification (iTRAQ) with two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS). 1,8-cineole was found to significantly damage the structure of Salmonella cell walls and membranes. A total of 3011 proteins were extracted from the experimental group, of which 435 were differentially expressed (1.5-fold) with 123 upregulated and 312 downregulated. The expressed proteins were involved in 935 intracellular biological processes, 98 cellular components, 477 molecular functions and 86 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among them, proteins associated with carbohydrate, nucleotide, amino acid, lipid, and energy metabolism were significantly changed following treatment with 1,8-cineole. Carbohydrate metabolism and membrane protein-related genes was down-regulated at the mRNA level when Salmonella was treated with 1,8-cineole. 1,8-cineole may be a potential antibiotic for Salmonella infections.

SUBMITTER: Sun Y

PROVIDER: S-EPMC5992416 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Effects of 1,8-cineole on Carbohydrate Metabolism Related Cell Structure Changes of Salmonella.

Sun Yangying Y Cai Xiaojun X Cao Jinxuan J Wu Zhen Z Pan Daodong D

Frontiers in microbiology 20180525

Salmonella is gram-negative foodborne zoonotic bacteria which triggers disease in humans. Our previous studies showed 1,8-cineole possessed remarkable antimicrobial effects on foodborne zoonotic bacteria indicating it could serve as a new source of antibiotic for Salmonella. Present study elucidated the antibacterial mechanism of 1,8-cineole by analyzing serum protein expressed by Salmonella following treatment with 1,8-cineole (0.25 mg/mL, 3 h) using isobaric tags for relat ...[more]

PMID: 29910778

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Project description:We report the isolation and characterization of three new cytochrome P450 monooxygenases: CYP101J2, CYP101J3, and CYP101J4. These P450s were derived from Sphingobium yanoikuyae B2, a strain that was isolated from activated sludge based on its ability to fully mineralize 1,8-cineole. Genome sequencing of this strain in combination with purification of native 1,8-cineole-binding proteins enabled identification of 1,8-cineole-binding P450s. The P450 enzymes were cloned, heterologously expressed (N-terminally His6 tagged) in Escherichia coli BL21(DE3), purified, and spectroscopically characterized. Recombinant whole-cell biotransformation in E. coli demonstrated that all three P450s hydroxylate 1,8-cineole using electron transport partners from E. coli to yield a product putatively identified as (1S)-2α-hydroxy-1,8-cineole or (1R)-6α-hydroxy-1,8-cineole. The new P450s belong to the CYP101 family and share 47% and 44% identity with other 1,8-cineole-hydroxylating members found in Novosphingobium aromaticivorans and Pseudomonas putida Compared to P450cin (CYP176A1), a 1,8-cineole-hydroxylating P450 from Citrobacter braakii, these enzymes share less than 30% amino acid sequence identity and hydroxylate 1,8-cineole in a different orientation. Expansion of the enzyme toolbox for modification of 1,8-cineole creates a starting point for use of hydroxylated derivatives in a range of industrial applications.ImportanceCYP101J2, CYP101J3, and CYP101J4 are cytochrome P450 monooxygenases from S. yanoikuyae B2 that hydroxylate the monoterpenoid 1,8-cineole. These enzymes not only play an important role in microbial degradation of this plant-based chemical but also provide an interesting route to synthesize oxygenated 1,8-cineole derivatives for applications as natural flavor and fragrance precursors or incorporation into polymers. The P450 cytochromes also provide an interesting basis from which to compare other enzymes with a similar function and expand the CYP101 family. This could eventually provide enough bacterial parental enzymes with similar amino acid sequences to enable in vitro evolution via DNA shuffling.

Project description:Chlorhexidine belongs to a group of medicines called antiseptic antibacterial agents. Chlorhexidine is commonly used for the care and clean off the skin, hands, and wounds. In recent years, medicinal and aromatic plants have been used for prevention of disease, maintaining health, and improving disease in traditional and modern medicine as a medicament. According to recent research, cineole is the isolated active agent of eucalyptus oil and possesses antimicrobial activity. It was demonstrated that cineole could enhance the antimicrobial effects of the other antiseptics.The aim of this study was to investigate the efficacy of 1,8-cineole on the antimicrobial effect of chlorhexidine against some microorganisms.The effect of 1,8-cineole on antimicrobial activity of chlorhexidine gluconate (CHG) was tested using seven different microorganisms. In this study, CHG (128-0.125 mg/l) and cineole (512-2 g/l) were analyzed together and separately using checkerboard assay. Interactions between CHG and 1,8-cineole have been identified as synergistic, indifferent, or antagonistic.Synergistic activity was demonstrated between CHG and 1,8-cineole against Staphylococcus aureus, methicillin-resistant S. aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Candida albicans. Indifferent interactions for these compounds were demonstrated against Pseudomonas aeruginosa.CHG antiseptic properties were found to be increased when CHG was used in combination with 1,8-cineole. In this way, CHG will reveal stronger effect against microorganisms.Cineole has increased the antimicrobial activity of chlorhexidine gluconate against all microorganisms except Pseudomonas aeruginosaIn topical application, using cineole in combination with chlorhexidine may be easier, eradicate certain resistant bacteria by increasing the antimicrobial efficacy. Abbreviation Used: CHG: Chlorhexidine gluconate, MRSA: Methicillin-resistant S. aureus, MHB: Mueller Hinton broth, SDB: Sabouraud dextrose broth, CFU: Colonyforming unit, FIC: Fractional inhibitory concentration, FICI: FIC index, EO: Eucalyptus oil.

Dataset Information

Effects of 1,8-cineole on Carbohydrate Metabolism Related Cell Structure Changes of Salmonella.

Publications

Effects of 1,8-cineole on Carbohydrate Metabolism Related Cell Structure Changes of <i>Salmonella</i>.

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