Cloning, sequencing and expression of the gene encoding the extracellular neutral protease, vibriolysin, of Vibrio proteolyticus

David, Victor A.; Deutch, A H; Sloma, Alan; Pawlyk, Diane M.; Durham, D R

doi:10.1016/0378-1119(92)90310-l

Cited by 65 publications

(29 citation statements)

References 24 publications

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“…The predicted amino acid sequence of A. hydrophila AhpB showed homology with several metalloproteases from Vibrio spp. (9,11,16,33), Helicobacter pylori hemagglutinin/proteinase fragment (46), Vibrio cholerae hemagglutinin/proteinase precursor (16), and P. aeruginosa elastase precursor (LasB) (14) (Fig. 3).…”

Section: Molecular Cloning Of Thementioning

confidence: 99%

A Major Secreted Elastase Is Essential for Pathogenicity of Aeromonas hydrophila

et al. 2000

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Aeromonas hydrophila is an opportunistic pathogen and the leading cause of fatal hemorrhagic septicemia in rainbow trout. A gene encoding an elastolytic activity, ahyB, was cloned from Aeromonas hydrophila AG2 into pUC18 and expressed in Escherichia coli and in the nonproteolytic species Aeromonas salmonicida subsp. masoucida. Nucleotide sequence analysis of the ahyB gene revealed an open reading frame of 1,764 nucleotides with coding capacity for a 588-amino-acid protein with a molecular weight of 62,728. The first 13 N-terminal amino acids of the purified protease completely match those deduced from DNA sequence starting at AAG (Lys-184). This finding indicated that AhyB is synthesized as a preproprotein with a 19-amino-acid signal peptide, a 164-amino-acid N-terminal propeptide, and a 405-amino-acid intermediate which is further processed into a mature protease and a C-terminal propeptide. The protease hydrolyzed casein and elastin and showed a high sequence similarity to other metalloproteases, especially with the mature form of the Pseudomonas aeruginosa elastase (52% identity), Helicobacter pylori zinc metalloprotease (61% identity), or proteases from several species of Vibrio (52 to 53% identity). The gene ahyB was insertionally inactivated, and the construct was used to create an isogenic ahyB mutant of A. hydrophila. These first reports of a defined mutation in an extracellular protease of A. hydrophila demonstrate an important role in pathogenesis.Aeromonas hydrophila is a gram-negative opportunistic pathogen in humans and several fish species, causing soft tissue wound infections and diarrhea in the former (1, 18, 21) and fatal hemorrhagic septicemia in the latter (2,12,15,37). It has been speculated that A. hydrophila virulence could involve several extracellular enzymes including proteases, hemolysins, enterotoxins, and acetylcholinesterase. Some of the toxins have been biochemically characterized, but their precise roles in the pathogenicity of A. hydrophila have not yet been determined (8,29,35,41,42). The two major extracellular proteolytic activities of A. hydrophila that have been described so far, a 38-kDa thermostable metalloprotease (29, 41) and a 68-kDa temperature-labile serine protease (30,42), are present in most A. hydrophila culture supernatants. In addition, a 19-kDa zinc proteinase was found in the growth medium of a strain of A. hydrophila isolated from the intestinal tract of the leech Hirudo medicinalis (31), and a 22-kDa serine proteinase, which is stable at 56°C for 10 min, was purified from A. hydrophila strain B 32 culture supernatant (43). Several strategies have been used to examine the role of some A. hydrophila proteases in virulence, including Tn5-induced protease-deficient mutants of A. hydrophila (29) and direct inoculation of purified 22-kDa serine protease in rainbow trout (43), but with conflicting results. Two major secretion products of A. salmonicida, an extracellular serine protease (AspA) and a glycerophospholipid:cholesterol acyltransferase (SatA), had previous...

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Section: Molecular Cloning Of Thementioning

confidence: 99%

A Major Secreted Elastase Is Essential for Pathogenicity of Aeromonas hydrophila

et al. 2000

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“…Our preliminary experiments using partially purified preparations indicated that an extracellular protease produced by biotype 2 might be indistinguishable from VVP. In spite of their being avirulent to humans, other vibrios such as V. anguillarum [8,9] and V. proteolyticus [10] also produce closely related metalloproteases those exhibit 70-80 % identities in the primary structures and have comparable biological activities. Additionally, these proteolytic enzymes are neutralized with the antibody against VVP [11].…”

Section: Introductionmentioning

confidence: 99%

High growing ability of Vibrio vulnificus biotype 1 is essential for production of a toxic metalloprotease causing systemic diseases in humans

Watanabe

Miyoshi

Kawase

et al. 2004

Microbial Pathogenesis

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Vibrio vulnificus biotype 1, a causative agent of fatal septicemia or wound infection in humans, is known to produce a toxic metalloprotease as an important virulence determinant. V. vulnificus biotype 2 (serovar E), a primary eel pathogen, was found to elaborate an extracellular metalloprotease that was indistinguishable from that of biotype 1. The potential of V. vulnificus biotype 1 for production of the metalloprotease was compared with biotype 2 and other human non-pathogenic Vibrio species (V. anguillarum and V. proteolyticus). When cultivated at 25o C in tryptone-yeast extract broth supplemented with 0.9 % NaCl, all bacteria multiplied sufficiently and secreted significant amounts of the metalloprotease. However, at 37 o C with 0.9 % NaCl, V.anguillarum neither grew nor produced the metalloprotease. In human serum, only V.vulnificus biotype 1 revealed a steady multiplication accompanied with production of the extracellular metalloprotease. This prominent ability of biotype 1 in growth and protease production may contribute to cause serious systemic diseases in humans.

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“…The empI gene encoded a putative protein of 731 amino acids with a molecular weight of 77,784. A computer-assisted sequence similarity search revealed that a 304-amino-acid EmpI protein starting with Ala-207 showed 61.2, 63.0, and 64.4z identity with the mature P. aeruginosa elastase, 9) the V. vulniˆcus metal protease, 10) and the V. proteolyticus vibriolysin, 11) respectively. These proteases are zinc metalloproteases.…”

mentioning

confidence: 99%

Cloning and Characterization of Extracellular Metal Protease Gene of the Algicidal Marine BacteriumPseudoalteromonassp. Strain A28

Lee

Kato

Nakashima

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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Cloning, sequencing and expression of the gene encoding the extracellular neutral protease, vibriolysin, of Vibrio proteolyticus

Cited by 65 publications

References 24 publications

A Major Secreted Elastase Is Essential for Pathogenicity of Aeromonas hydrophila

A Major Secreted Elastase Is Essential for Pathogenicity of Aeromonas hydrophila

High growing ability of Vibrio vulnificus biotype 1 is essential for production of a toxic metalloprotease causing systemic diseases in humans

Cloning and Characterization of Extracellular Metal Protease Gene of the Algicidal Marine BacteriumPseudoalteromonassp. Strain A28

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