Influence of growth conditions on diverse polysaccharide production by<i>Campylobacter jejuni</i>

Corcoran, Adrian T.; Moran, Anthony P.

doi:10.1111/j.1574-695x.2006.00178.x

Cited by 14 publications

(15 citation statements)

References 41 publications

(71 reference statements)

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“…This is consistent with previous observations that C. jejuni is capable of producing a variety of polysaccharide-related structures that are influenced by growth conditions, such as temperature [26]. …”

Section: Discussionsupporting

confidence: 93%

Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

et al. 2010

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BackgroundCampylobacter jejuni is a major bacterial cause of food-borne enteritis, and its lipooligosaccharide (LOS) plays an initiating role in the development of the autoimmune neuropathy, Guillain-Barré syndrome, by induction of anti-neural cross-reactive antibodies through ganglioside molecular mimicry.ResultsHerein we describe the existence and heterogeneity of multiple LOS forms in C. jejuni strains of human and chicken origin grown at 37°C and 42°C, respectively, as determined on sodium dodecyl sulphate-polyacrylamide electrophoresis gels with carbohydrate-specific silver staining and blotting with anti-ganglioside ligands, and confirmed by nuclear magnetic resonance (NMR) spectroscopy. The C. jejuni NCTC 11168 original isolate (11168-O) was compared to its genome-sequenced variant (11168-GS), and both were found to have a lower-Mr LOS form, which was different in size and structure to the previously characterized higher-Mr form bearing GM1 mimicry. The lower-Mr form production was found to be dependent on the growth temperature as the production of this form increased from ~5%, observed at 37°C to ~35% at 42°C. The structure of the lower-Mr form contained a β-D-Gal-(1→3)-β-D-GalNAc disaccharide moiety which is consistent with the termini of the GM1, asialo-GM1, GD1, GT1 and GQ1 gangliosides, however, it did not display GM1 mimicry as assessed in blotting studies but was shown in NMR to resemble asialo-GM1. The production of multiple LOS forms and lack of GM1 mimicry was not a result of phase variation in the genes tested of NCTC 11168 and was also observed in most of the human and chicken isolates of C. jejuni tested.ConclusionThe presence of differing amounts of LOS forms at 37 and 42°C, and the variety of forms observed in different strains, indicate that LOS form variation may play a role in an adaptive mechanism or a stress response of the bacterium during the colonization of different hosts.

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Section: Discussionsupporting

confidence: 93%

Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

et al. 2010

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“…Finally, our observations suggest that biofilm formation and the production of the CFW-reactive polysaccharide may represent important C. jejuni stress responses induced under adverse conditions such as those encountered during extended growth and in the absence of an SR. For instance, although C. jejuni stationary phase is typically associated with detrimental or poorly understood effects such as oxidative stress, peptidoglycan and metabolism changes, and conversion to coccoid and viable-but-nonculturable forms (31,74,75), it has recently been shown to elicit general increases and alterations in polysaccharide composition (18,52), similar to observations for other bacteria (9,14,43). Our stress response hypothesis is also consistent with the dim13 mutant serum sensitivity defect, the dim10 mutant late-stage culturability defect (which may reflect rapid nutrient depletion and toxic metabolite production without concomitant up-regulation of the protective polysaccharide), and a recent study describing the altered expression of oxidative and stress response proteins in biofilm versus planktonic populations of C. jejuni 11168 (36).…”

Section: Discussionmentioning

confidence: 99%

Campylobacter jejuniBiofilms Up-Regulated in the Absence of the Stringent Response Utilize a Calcofluor White-Reactive Polysaccharide

et al. 2008

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The enteric pathogen Campylobacter jejuni is a highly prevalent yet fastidious bacterium. Biofilms and surface polysaccharides participate in stress survival, transmission, and virulence in C. jejuni; thus, the identification and characterization of novel genes involved in each process have important implications for pathogenesis. We found that C. jejuni reacts with calcofluor white (CFW), indicating the presence of surface polysaccharides harboring ␤1-3 and/or ␤1-4 linkages. CFW reactivity increased with extended growth, under 42°C anaerobic conditions, and in a ⌬spoT mutant defective for the stringent response (SR). Conversely, two newly isolated dim mutants exhibited diminished CFW reactivity as well as growth and serum sensitivity differences from the wild type. Genetic, biochemical, and nuclear magnetic resonance analyses suggested that differences in CFW reactivity between wild-type and ⌬spoT and dim mutant strains were independent of well-characterized lipooligosaccharides, capsular polysaccharides, and N-linked polysaccharides. Targeted deletion of carB downstream of the dim13 mutation also resulted in CFW hyporeactivity, implicating a possible role for carbamoylphosphate synthase in the biosynthesis of this polysaccharide. Correlations between biofilm formation and production of the CFW-reactive polymer were demonstrated by crystal violet staining, scanning electron microscopy, and confocal microscopy, with the C. jejuni ⌬spoT mutant being the first SR mutant in any bacterial species identified as up-regulating biofilms. Together, these results provide new insight into genes and processes important for biofilm formation and polysaccharide production in C. jejuni.

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“…Batch culture remains an extremely common method of growing C. jejuni in vitro and is itself a common experimental system (Corcoran & Moran, 2007;Mohammed et al, 2005). In light of this, in addition to informing the design and interpretation of studies that employ C. jejuni grown in vitro, greater understanding of the growth cycle presents a sensible aim in itself.…”

Section: Introductionmentioning

confidence: 99%

Metabolite and transcriptome analysis of Campylobacter jejuni in vitro growth reveals a stationary-phase physiological switch

et al. 2009

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Campylobacter jejuni is a prevalent cause of food-borne diarrhoeal illness in humans. Understanding of the physiological and metabolic capabilities of the organism is limited. We report a detailed analysis of the C. jejuni growth cycle in batch culture. Combined transcriptomic, phenotypic and metabolic analysis demonstrates a highly dynamic 'stationary phase', characterized by a peak in motility, numerous gene expression changes and substrate switching, despite transcript changes that indicate a metabolic downshift upon the onset of stationary phase. Video tracking of bacterial motility identifies peak activity during stationary phase. Amino acid analysis of culture supernatants shows a preferential order of amino acid utilization. Proton NMR ( 1 H-NMR) highlights an acetate switch mechanism whereby bacteria change from acetate excretion to acetate uptake, most probably in response to depletion of other substrates. Acetate production requires pta (Cj0688) and ackA (Cj0689), although the acs homologue (Cj1537c) is not required. Insertion mutants in Cj0688 and Cj0689 maintain viability less well during the stationary and decline phases of the growth cycle than wild-type C. jejuni, suggesting that these genes, and the acetate pathway, are important for survival. INTRODUCTIONCampylobacter jejuni is the major cause of food-borne bacterial gastroenteritis worldwide, with an estimated 1 in 100 individuals in both the USA and the UK developing Campylobacter-related illness each year (Gaynor et al., 2004;Gillespie et al., 2002). Although most cases are selflimiting, C. jejuni can cause severe post-infection complications including the peripheral neuropathies GuillainBarré and Miller-Fisher syndromes (Nachamkin et al., 2000). Despite the significance of C. jejuni as a food-borne pathogen, our understanding of its basic biochemistry and physiology, gene regulation, colonization, virulence and environmental survival mechanisms lags behind that of many other pathogenic and non-pathogenic bacteria, and many questions regarding C. jejuni physiology and pathogenesis remain (Young et al., 2007). Research has been hampered by a lack of genetic tools, the difficulty of growing the bacteria in the laboratory and the absence of a suitable animal model that mimics human disease.C. jejuni is a fastidious bacterium, with a limited capacity for biosynthesis, and requires complex growth media (Kelly, 2001). Establishing the metabolic capabilities of C. jejuni is central to comprehension of environmental persistence and host colonization. The microaerophilic nature, complex nutritional requirements and relative difficulty of culturing C. jejuni have all contributed to the limited understanding of metabolism (Kelly, 2001(Kelly, , 2005Sellars et al., 2002). C. jejuni is unable to catabolize glucose and other hexose sugars due to the absence of the key glycolytic enzyme 6-phosphofructokinase (Parkhill et al., 2000;Velayudhan & Kelly, 2002 The importance of each amino acid in C. jejuni metabolism is yet to be fully established, however, and i...

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Influence of growth conditions on diverse polysaccharide production byCampylobacter jejuni

Cited by 14 publications

References 41 publications

Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

Temperature-dependent phenotypic variation of Campylobacter jejuni lipooligosaccharides

Campylobacter jejuniBiofilms Up-Regulated in the Absence of the Stringent Response Utilize a Calcofluor White-Reactive Polysaccharide

Metabolite and transcriptome analysis of Campylobacter jejuni in vitro growth reveals a stationary-phase physiological switch

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