Regulation of Extracellular Alkaline Protease Activity by Histidine in a Collagenolytic Vibrio alginolyticus Strain

Long, Susan K. De; Mothibeli, M. A.; Robb, Frank T.; Woods, David R.

doi:10.1099/00221287-127-1-193

Cited by 39 publications

(43 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, we found that the deduced protein sequence of asp showed close homology to those of V. alginolyticus strain 12G01 and V. parahaemolyticus strain RIMD 2210633, but not to that of V. alginolyticus strain HY9901. These were not unexpected results because there was evidence that the extracellular alkaline serine proteases produced by V. alginolyticus might differ greatly in their sequences, depending on the strain, source of the isolate, and the medium [1,8,12,19,30]. Phenylmethylsulfonyl fluoride (PMSF) is a covalently binding irreversible serine protease inhibitor.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Role of Alkaline Serine Protease, Asp, in Vibrio alginolyticus Virulence and Regulation of Its Expression by LuxO-LuxR Regulatory System

Rui

Liu²,

Wang³

et al. 2009

J.Microbiol.Biotechnol

View full text Add to dashboard Cite

The alkaline serine protease asp, which was shown to be a virulence factor of Vibrio alginolyticus as a purified protein, was cloned from V. alginolyticus EPGS, a strain recently isolated from moribund Epinephelus coioides in an outbreak of vibriosis in a mariculture farm of Shenzhen. The asp null mutant was constructed by homologous recombination with suicide plasmid pNQ705-1. Compared with the wild-type strain, the asp null mutant exhibited a significant decrease of total extracellular protease activity, and caused a 15-fold decrease in virulence of V. alginolyticus. In our previous study, the luxO and luxR val genes from V. alginolyticus MVP01 were cloned and identified, and the luxO-luxR val regulatory couple was shown to regulate various genes expression, suggesting that it played a central role in the quorum sensing system of V. alginolyticus. In this study, the regulation of the asp gene was analyzed by using RT-PCR and quantitative real-time PCR methods; we proved that its transcription was greatly induced at the late stage of growth and was regulated by a luxO-luxR val regulatory system. Keywords: Alkaline serine protease, asp, quorum sensing, Vibrio alginolyticus Quorum sensing, a type of cell-cell signaling, has been studied widely in recent years [3]. A typical quorum sensing system of Gram-negative bacteria has two types of autoinducers: acylated homoserine lactones (AI-1) and furanosyl borate diester (AI-2) [4,28]. In Vibrio harveyi, detection of and response to AI-1 and AI-2 occurs through two parallel two-component signal transduction circuits, and a shared response regulator called LuxO integrates the information from these two circuits and relays it to LuxR, whereupon activated LuxR regulates the expression of various genes [20]. Vibrio spp. could use this system to control many processes including bioluminescence, virulence, mobility, biofilm formation, and extracellular polysaccharide production [32]. Significantly, extracellular proteases, the virulence factors of Vibrio spp., were shown to be regulated by their respective quorum sensing systems, such as hemagglutinin protease hap in V. cholerae [13], metalloprotease empA in V. anguillarum [6], and metalloprotease vvp and cytolysin vvhA in V. vulnificus [25].Vibrio alginolyticus, a ubiquitous organism in seawater, has been isolated from different organisms as part of the normal marine flora. However, it has also been suggested that this species was a pathogen of several marine animals and humans. In the South China Sea, V. alginolyticus was reported to be the dominant causative agent of highmortality vibriosis in the large yellow croaker, sea bream, grouper, kuruma prawn, as well as shellfish species [16,17].Compared with other Vibrio spp., such as V. cholerae, V. anguillarum, and V. vulnificus, much less is known about the pathogenic mechanism of V. alginolyticus. Studies on the pathogenicity of various isolates of V. alginolyticus suggested that some extracellular enzymes with strong proteolytic activities could be important virulent facto...

show abstract

Section: Discussionmentioning

confidence: 99%

“…because there was evidence that the extracellular alkaline serine proteases might be degraded as they were secreted from the cytoplasm [1,12,14,19].…”

Section: Quorum Sensing Regulates Asp Expression 435mentioning

confidence: 99%

Role of Alkaline Serine Protease, Asp, in Vibrio alginolyticus Virulence and Regulation of Its Expression by LuxO-LuxR Regulatory System

Rui

Liu²,

Wang³

et al. 2009

J.Microbiol.Biotechnol

View full text Add to dashboard Cite

show abstract

“…In the presence of collagen or macromolecular fragments of collagen (15), a subculture of this organism (strain 1.029) produces a collagenase having a very high specific activity (12,20). The structural and functional properties of this Achromobacter collagenase (EC 3.4.24.8), as well as the metabolism of strain 1.029, have been described in a series of papers by Woods et al (21,26,27,29,35) and by workers in our laboratory (4, 10,11,13,14,19,30).…”

mentioning

confidence: 99%

Assignment of "Achromobacter iophagus" Strain I.029 to Vibrio alginolyticus Chemovar iophagus

Emod

Soubigou

Tong

et al. 1983

International Journal of Systematic Bacteriology

View full text Add to dashboard Cite

RICHARD^Strain CIP 82.01 of Vibrio alginolyticus and strain 1.029 (previously designated "Achromobacter iophagus") were compared. Strain 1.029 produces a collagenase of high specific activity (Achrornobacter collagenase; EC 3.4.24.8). Collagenase production is induced in strain CIP 82.01 by collagen or macromolecular fragments of collagen in a manner similar to collagenase induction in strain 1.029; caseinolytic proteinase is constitutive. In this study we demonstrate that both strains also produce a constitutive extracellular endonuclease. Collagenases from both strains cleave either native collagen in its helical region or a similar synthetic peptide; both enzymes are inhibited by ethylenediamine tetraacetate, but not by diisopropyl fluorophosphate. The collagenase subunit (molecular weight, 35,000) of strain CIP 82.01 is similar in amino acid composition to the subunit of the strain 1.029 enzyme, although some of the aspartic and threonine residues in strain CIP 82.01 are replaced by glutamic and serine residues in strain 1.029. Surface radioiodination followed by two-dimensional electrophoresis showed that there are quantitative differences in the major outer membrane proteins of the two strains. Strains CIP 82.01 and 1.029 differ qualitatively in resistance to ampicillin and carbenicillin, in cellobiose fermentation, in ornithine decarboxylase activity, and in halophilism. We propose that strain 1.029, which was originally designated A . iophagus, be included within the species V. alginolyticus, but that this organism be distinguished from other strains of this species by the designation Vibrio alginolyticus chemovar iophagus, with the corresponding collagenase designated "iophagus collagenase" (EC 3.4.24.8).In 1973, Welton and Woods isolated and described the gram-negative aerobic bacterium Achromobacter iophagus (34). In the presence of collagen or macromolecular fragments of collagen (15), a subculture of this organism (strain 1.029) produces a collagenase having a very high specific activity (12,20). The structural and functional properties of this Achromobacter collagenase (EC 3.4.24.8), as well as the metabolism of strain 1.029, have been described in a series of papers by Woods et al. (21,26,27,29,35) and by workers in our laboratory (4,10,11,13,14,19,30).In a recent study (27), which confirmed the original observation of collagenase induction by macromolecular collagen fragments (15), Achromobacter iophagus was reclassified as a strain of Vibrio alginolyticus; however, a direct experimental comparison of strain 1.029 with a welldefined wild-type strain of V . alginolyticus has been lacking. It is well established that the other strains of the Vibrionaceae (23, 31) can be induced (by collagen and fragments of collagen) to produce collagenolytic and gelatinolytic enzymes.We undertook a comparative study to determine the differences between strain 1.029 isolated from a random selection of cured hides in South Africa (34) and a wild-type strain (strain CIP 82.01) of V . alginolyticus from a different ge...

show abstract

“…Azocasein (Sigma) (for substrate suitability, see Reimerdes & Klostermoyer 1976) was made up of 0.5 "10 in 0.2 M Tris-HC1 pH 7.8, and the protcase assayed after the method of Long et al (1981). Then 250 1-11 enzyme was added to 500 ~1 1 of azocasein substrate and incubated in a shaking water bath at 20°C for 0, 1 and 3 h. The reaction was stopped by the addition of 1 m1 10 % trichloracetic acid and the vials were held at 5°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Digestive enzymes of the anchovy Engraulis capensis in relation to diet

Seiderer¹,

Davis²,

Robb³

et al. 1987

Mar. Ecol. Prog. Ser.

View full text Add to dashboard Cite

Digestive enzymes of the anchovy Engraulis capensis in relation to diet' Department of Microbiology, University of Cape Town, Rondebosch 7700, South Africa Institute for Marine Environmental Research, Prospect Place, Plymouth PL1 3DH, United Kingdom ABSTRACT: The anchovy Engraulis capensis (Glch.) is capable of exploiting both diatoms and Crustacea as a food resource. Much of the crustacean exoskeletal rnatenal passes into the hindgut after initial dsruphon in the stomach. Both laminarinase and cl-amylase activity are relatively low in the oesophagus and stomach, but increase in the caeca and lumen of the intestine. There is no evidence of cellulolytic activity in the gut although numerous gut bacteria occur in the caeca and lumen of the intestine. Distribution of protease activity in the gut follows that of carbohydrases, most proteolytic activity being present in the lumen of the intestine, rather than in the oesophagus or stomach. In contrast, chitinase activity is found early in the digestive pathway and is capable of attacking the chitinous exoskeleton of Crustacea in the stomach, prior to hydrolysis of their contents by carbohydrases and proteases. Bacteria isolated from the intestinal caeca do not hydrolyse chitin, nor do they degrade long-chain polysaccharides or azocasein. Most of the isolates were capable of utilisation of simple sugars cornnlonly found in diatoms and released following hydrolysis of polysaccharides, and also of utilisation of N-acetylglucosamine formed as an endproduct of chitin hydrolysis. Anaerobic strains were, In addition, capable of urea utilisation. Our data thus suggest that digestive enzymes allow E capensjs to exploit a wide range of food items, including diatoms and Crustacea, without bacterial involvement. The gut rnicroflora may be primarily maintained by uptake of the endproducts of carbohydrate, protein and chitin degradation.

show abstract

Regulation of Extracellular Alkaline Protease Activity by Histidine in a Collagenolytic Vibrio alginolyticus Strain

Cited by 39 publications

References 10 publications

Role of Alkaline Serine Protease, Asp, in Vibrio alginolyticus Virulence and Regulation of Its Expression by LuxO-LuxR Regulatory System

Role of Alkaline Serine Protease, Asp, in Vibrio alginolyticus Virulence and Regulation of Its Expression by LuxO-LuxR Regulatory System

Assignment of "Achromobacter iophagus" Strain I.029 to Vibrio alginolyticus Chemovar iophagus

Digestive enzymes of the anchovy Engraulis capensis in relation to diet

Contact Info

Product

Resources

About