Expression, purification, and characterization of thermolabile hemolysin (TLH) from Vibrio alginolyticus

Jia, Airong; Woo, Norman Y.S.; Zhang, Xiao‐Hua

doi:10.3354/dao02225

Cited by 32 publications

(20 citation statements)

References 29 publications

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“…This gene cluster is conserved in 24 Pseudoalteromonas genomes distributed across the phylogenetic tree (Supplementary Figure S1 and Supplementary Table S7 ). Similar potential virulence clusters are also found in the genome of the pathogenic Gammaproteobacteria Vibrio alginolyticus 12G01, from which the thermostable hemolysin was characterized and shown to be toxic to zebrafish ( Jia et al, 2010 ), and of the Alphaproteobacteria Nautella sp. R11, a red algal pathogen ( Fernandes et al, 2011 ).…”

Section: Resultsmentioning

confidence: 57%

Evolutionary Evidence of Algal Polysaccharide Degradation Acquisition by Pseudoalteromonas carrageenovora 9T to Adapt to Macroalgal Niches

et al. 2018

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About half of seaweed biomass is composed of polysaccharides. Most of these complex polymers have a marked polyanionic character. For instance, the red algal cell wall is mainly composed of sulfated galactans, agars and carrageenans, while brown algae contain alginate and fucose-containing sulfated polysaccharides (FCSP) as cell wall polysaccharides. Some marine heterotrophic bacteria have developed abilities to grow on such macroalgal polysaccharides. This is the case of Pseudoalteromonas carrageenovora 9T (ATCC 43555T), a marine gammaproteobacterium isolated in 1955 and which was an early model organism for studying carrageenan catabolism. We present here the genomic analysis of P. carrageenovora. Its genome is composed of two chromosomes and of a large plasmid encompassing 109 protein-coding genes. P. carrageenovora possesses a diverse repertoire of carbohydrate-active enzymes (CAZymes), notably specific for the degradation of macroalgal polysaccharides (laminarin, alginate, FCSP, carrageenans). We confirm these predicted capacities by screening the growth of P. carrageenovora with a large collection of carbohydrates. Most of these CAZyme genes constitute clusters located either in the large chromosome or in the small one. Unexpectedly, all the carrageenan catabolism-related genes are found in the plasmid, suggesting that P. carrageenovora acquired its hallmark capacity for carrageenan degradation by horizontal gene transfer (HGT). Whereas P. carrageenovora is able to use lambda-carrageenan as a sole carbon source, genomic and physiological analyses demonstrate that its catabolic pathway for kappa- and iota-carrageenan is incomplete. This is due to the absence of the recently discovered 3,6-anhydro-D-galactosidase genes (GH127 and GH129 families). A genomic comparison with 52 Pseudoalteromonas strains confirms that carrageenan catabolism has been recently acquired only in a few species. Even though the loci for cellulose biosynthesis and alginate utilization are located on the chromosomes, they were also horizontally acquired. However, these HGTs occurred earlier in the evolution of the Pseudoalteromonas genus, the cellulose- and alginate-related loci being essentially present in one large, late-diverging clade (LDC). Altogether, the capacities to degrade cell wall polysaccharides from macroalgae are not ancestral in the Pseudoalteromonas genus. Such catabolism in P. carrageenovora resulted from a succession of HGTs, likely allowing an adaptation to the life on the macroalgal surface.

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Section: Resultsmentioning

confidence: 57%

Evolutionary Evidence of Algal Polysaccharide Degradation Acquisition by Pseudoalteromonas carrageenovora 9T to Adapt to Macroalgal Niches

et al. 2018

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“…Consistent with these findings, none of the reference isolates of the various groups was found to carry any of these determinants. In contrast to those results for other virulence genes, a PCR for detection of the thermolabile hemolysin ( tlh ) gene showed that all SD isolates, as well as reference strains, have tlh , a gene playing a potentially key role in virulence associated with aquatic organisms and possibly humans as well (Wang et al., 2007; Jia et al., 2010).…”

Section: Resultsmentioning

confidence: 99%

Genetic and phylogenetic evidence for horizontal gene transfer among ecologically disparate groups of marine Vibrio

Hoffmann¹,

Monday²,

McCarthy³

et al. 2012

Cladistics

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Vibrio represents a diverse bacterial genus found in different niches of the marine environment, including numerous genera of marine sponges (phylum Porifera), inhabiting different depths and regions of benthic seas, that are potentially important in driving adaptive change among Vibrio spp. Using 16S rRNA gene sequencing, a previous study showed that sponge‐derived (SD) vibrios clustered with their mainstream counterparts present in shallow, coastal ecosystems, suggesting a genetic relatedness between these populations. Sequences from the topA, ftsZ, mreB, rpoD, rctB and toxR genes were used to investigate the degree of relatedness existing between these two separate populations by examining their phylogenetic and genetic disparity. Phylogenies were constructed from the concatenated sequences of the six housekeeping genes using maximum‐parsimony, maximum‐likelihood and neighbour‐joining algorithms. Genetic recombination was evaluated using the incongruence length difference test, Split decomposition and measuring overall compatibility of sites. This combined technical approach provided evidence that SD Vibrio strains are largely genetically homologous to their shallow‐water counterparts. Moreover, the analyses conducted support the existence of extensive horizontal gene transfer between these two groups, supporting the idea of a single panmictic population structure among vibrios from two seemingly distinct, marine environments.

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“…We hypothesize that intrinsic factors in egg fried rice promote V. alginolyticus to produce a concentration of the hemolytic factor, related to the thermolabile hemolysin (TLH). Jia and co-workers [44] have shown that the TLH gene from V. alginolyticus shared 94 % identity with the lecithin-dependent hemolysin of V. parahaemolyticus which can damage the cell membranes including that of ounder red blood cells. Thus, we deduce that the lecithin in egg fried rice stimulates the V. alginolyticus TLH gene resulting in an increase in the hemolytic activity.…”

Section: Discussionmentioning

confidence: 99%

Effect of food matrix type on growth characteristics and hemolysin production of Vibrio alginolyticus

Deng

et al. 2020

Preprint

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Background: The growth and hemolysin production of V. alginolyticus at 30 °C in briny Tilapia , shrimp, scallop, oyster, pork, chicken, freshwater fish and egg fried rice were investigated. Bacterial counts were enumerated by plate counting. Hemolysin production was evaluated by blood agar and hemolytic titer tests. Results: Based on the goodness of fit primary model statistics (R 2 , MSE, BF, AF), the modified Gompertz model was a better fit to V. alginolyticus growth in foods than the logistic model. Growth kinetic parameters of V. alginolyticus displayed a higher μ max and shorter λ in briny Tilapia > shrimp > freshwater fish > egg fried rice > scallop > oyster > chicken > pork. It was notable that the V. alginolyticus counts were similar at the stationary phase, with no significant growth behavior difference between raw and cooked foods. However, higher thermostable direct hemolysin activity and hemolytic titer were observed in briny Tilapia > egg fried rice > shrimp > freshwater fish > chicken > scallop > oyster > pork. Conclusion: V. alginolyticus growth was good in all food matrix types tested. Contrary to current belief, V. alginolyticus displayed a higher hemolytic activity in some non-seafoods (freshwater fish, egg fried rice and chicken) than in scallop or oyster. This is the first report of growth and toxicity of V. alginolyticus in non-seafood. This finding will significantly improve the accuracy of microbial risk assessment of V. alginolyticus in different food matrices especially during warmer climatic periods when it is most prevalent.

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Expression, purification, and characterization of thermolabile hemolysin (TLH) from Vibrio alginolyticus

Cited by 32 publications

References 29 publications

Evolutionary Evidence of Algal Polysaccharide Degradation Acquisition by Pseudoalteromonas carrageenovora 9T to Adapt to Macroalgal Niches

Evolutionary Evidence of Algal Polysaccharide Degradation Acquisition by Pseudoalteromonas carrageenovora 9T to Adapt to Macroalgal Niches

Genetic and phylogenetic evidence for horizontal gene transfer among ecologically disparate groups of marine Vibrio

Effect of food matrix type on growth characteristics and hemolysin production of Vibrio alginolyticus

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