Differential gene expression and extracellular secretion of the collagenolytic enzymes by the pathogen Vibrio parahaemolyticus

Miyoshi, Shin Ichi; Nitanda, Yuko; Fujii, Kaori; Kawahara, Kiyomi; Li, Tao; Maehara, Yoko; Ramamurthy, Thandavarayan; Takeda, Yasuharu; Shinoda, Sumió

doi:10.1111/j.1574-6968.2008.01159.x

Cited by 29 publications

(25 citation statements)

References 22 publications

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“…5A to D). Supporting these findings, the N-terminal amino acid sequencing analysis showed that the corresponding two forms of proteases observed in the supernatants isolated from early-stationary-phase cultures of V. parahaemolyticus were constituents of the VppC collagenase (63). Further, the processing of VchC was stimulated by a yet-to-be identified extracellular factor(s) (Fig.…”

Section: Discussionsupporting

confidence: 52%

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

et al. 2015

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bVibrio cholerae is autochthonous to various aquatic niches and is the etiological agent of the life-threatening diarrheal disease cholera. The persistence of V. cholerae in natural habitats is a crucial factor in the epidemiology of cholera. In contrast to the well-studied V. cholerae-chitin connection, scarce information is available about the factors employed by the bacteria for the interaction with collagens. Collagens might serve as biologically relevant substrates, because they are the most abundant protein constituents of metazoan tissues and V. cholerae has been identified in association with invertebrate and vertebrate marine animals, as well as in a benthic zone of the ocean where organic matter, including collagens, accumulates. Here, we describe the characterization of the V. cholerae putative collagenase, VchC, encoded by open reading frame VC1650 and belonging to the subfamily M9A peptidases. Our studies demonstrate that VchC is an extracellular collagenase degrading native type I collagen of fish and mammalian origin. Alteration of the predicted catalytic residues coordinating zinc ions completely abolished the protein enzymatic activity but did not affect the translocation of the protease by the type II secretion pathway into the extracellular milieu. We also show that the protease undergoes a maturation process with the aid of a secreted factor(s). Finally, we propose that V. cholerae is a collagenovorous bacterium, as it is able to utilize collagen as a sole nutrient source. This study initiates new lines of investigations aiming to uncover the structural and functional components of the V. cholerae collagen utilization program. Members of the genus Vibrio are important inhabitants of the marine coastal waters, estuaries, and ocean sediments, with some pathogenic species also causing wound infections, primary septicemia, gastroenteritis, and diarrhea in humans (1, 2). Among the more than 200 serogroups of V. cholerae, only O1 and O139 are recognized as agents of cholera, a potentially life-threatening diarrheal disease. Cholera remains a significant public health problem by affecting millions of people annually, predominantly in developing countries that have limited clean water supplies and poor sanitation (3). Infection of the human host occurs upon ingestion of water or food contaminated with the bacterium. Following colonization of the small intestine, V. cholerae utilizes a multicomponent secretory pathway, the type II secretion system (T2SS), to release cholera toxin. Cholera toxin induces acute diarrhea in people infected with cholera, enabling the bacteria to escape from the host into the aquatic reservoir (4). During interepidemic periods, V. cholerae persists freely or in association with various aquatic organisms, including copepods, insect egg masses, shellfish, and vertebrate fish (5-7). Extracellular proteins, including those secreted by the T2SS, have been implicated in facilitating the fitness of V. cholerae in both the human host and aquatic niches (4,6,8). The array of extrace...

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Section: Discussionsupporting

confidence: 52%

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

et al. 2015

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“…V. parahaemolyticus is a leading cause of bacterial gastroenteritis in Asia as well as in other countries Wong et al 2000;McLaughlin et al 2005;Shigeaki et al 2012), and is attracting increasing interest worldwide where raw or undercooked seafood is often consumed (Chao et al 2009). It causes wound infections in those exposed to contaminated water (Miyoshi et al 2008). It also causes septicemia, particularly in immunocompromised people .…”

Section: Introductionmentioning

confidence: 99%

Prevalence, antibiotic resistance, virulence and plasmid profiles of Vibrio parahaemolyticus from a tropical estuary and adjoining traditional prawn farm along the southwest coast of India

2015

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Vibrio parahaemolyticus, autochthonous to estuarine, marine, and coastal environments throughout the world, is a leading cause of foodborne gastroenteritis in Asia as well as in other countries. Considering their role in disease outbreaks, water and sediment samples from two different environments along the southwest coast of India, such as the Cochin estuary, which is a tourist hot spot and a traditional prawn farm, were examined for the detection of V. parahaemolyticus. A total of 120 presumptive vibrios were further screened using a chromogenic medium. The isolates producing the specific colour for V. parahaemolyticus on the chromogenic medium were confirmed by molecular methods. Using a PCR assay targeting the species-specific tlh gene, 75 isolates were confirmed as V. parahaemolyticus. The incidence of V. parahaemolyticus was found to be 71.6 % in the Cochin estuary and 53.3 % in the shrimp farm. The virulence genes tdh and trh were detected in two isolates from the Cochin estuary by multiplex PCR. Using 16S rRNA sequence analysis, our isolate exhibited 100 % similarity to the V. parahaemolyticus O3:K6 pandemic clone. The isolates were evaluated for their susceptibility towards 16 different antibiotics. All the isolates exhibited multiple antibiotic resistance (MAR). The MAR index of the isolates from the Cochin estuary ranged from 0.312 to 0.75 and that from the shrimp farm ranged from 0.1875 to 0.5. The plasmid profiles of the isolates were also checked. About 50.6 % of the isolates harboured plasmids and ten different profiles were observed. No correlation was observed between the antibiotic resistance patterns and plasmid profiles.

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“…The diversity of these enzymes is mainly correlated with the phylogenetic relationship of the three bacteria as follows: Clostridium, such as Clostridium histolyticum [7] and Clostridium perfringens [8]; Vibrio, such as Vibrio alginolyticus [9] and Vibrio parahaemolyticus [10]; and Porphyromonas, such as Porphyromonas gingivalis [11] and Porphyromonas endodontalis [12]. These microbes are pathogens or conditional pathogenic bacteria which potentially produce toxin.…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of a Novel Collagenase from Bacillus pumilus Col-J

et al. 2009

Appl Biochem Biotechnol

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The collagenase, produced extracellular by Bacillus pumilus Col-J, was purified by ammonium sulfate precipitation followed by two gel filtrations, involving Sephadex G-100 column and Sepharose Fast Flow column. Purified collagenase has a 31.53-fold increase in specific activity of 87.33 U/mg and 7.00% recovery. The collagenase has a relative molecular weight of 58.64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal temperature for the enzyme reaction was 45 degrees C. More than 50% of the original activity still remained after 5 min of incubation at 70 degrees C or 10 min at 60 degrees C. The maximal enzyme activity of collagenase was obtained at pH 7.5, and it was stable over a pH range of 6.5-8.0. The collagenase activity was strongly inhibited by Mn(2+), Pb(2+), ethylenediamine tetraacetic acid, ethylene glycol tetraacetic acid, and beta-mercaptoethanol. However, Ca(2+) and Mg(2+) greatly increased its activity. The collagenase from B. pumilus Col-J showed highly specific activity towards the native collagen from calf skin. The K(m) and V(max) of the enzyme for collagen were 0.79 mg/mL and 129.5 U, respectively.

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Differential gene expression and extracellular secretion of the collagenolytic enzymes by the pathogen Vibrio parahaemolyticus

Cited by 29 publications

References 22 publications

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

Prevalence, antibiotic resistance, virulence and plasmid profiles of Vibrio parahaemolyticus from a tropical estuary and adjoining traditional prawn farm along the southwest coast of India

Purification and Characterization of a Novel Collagenase from Bacillus pumilus Col-J

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