Purification and characterization of a marine bacterial collagenase

Merkel, Joseph R.; Dreisbach, Joseph H.

doi:10.1021/bi00607a025

Cited by 39 publications

(15 citation statements)

References 28 publications

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“…Besides, the gene encoding a collagenase has also been cloned and sequenced (36). Another Vibrio species, Vibrio harveyi (formerly called Vibrio B-30), produced two collagenases which have been purified and characterized; some properties of these enzymes are similar to those of C. histolyticum collagenase, such as molecular mass, specificity, and mode of collagen hydrolysis (26). In our previous study, the prtV gene, encoding a 62-kDa collagenase of Vibrio parahaemolyticus, was successfully cloned and expressed in Escherichia coli (22).…”

Section: Collagenasesmentioning

confidence: 99%

Metal Content and Biochemical Analyses of a Recombinant Collagenase PrtV from Vibrio parahaemolyticus

Yap

Lee

2000

Microbiology and Immunology

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PrtV is an extracellular metalloprotease of Vibrio parahaemolyticus and regarded as a collagenase. Inductively coupled plasma-optical emission spectrometry analysis indicated that the recombinant PrtV contains 1 mol of zinc per mol of the native enzyme. On the basis of a kinetic study using 2-furanacryloyl-LeuGly-Pro-Ala (FALGPA, the specific substrate for bacterial collagenase) as a substrate, it was suggested that metal ions may play a significant role in the binding and catalytic steps of the substrate. PrtV hydrolyzed type I, II, III, and IV collagens; however, it did not hydrolyze type V. In addition, the hydrolysis of native proteins and synthetic substrates revealed that PrtV possesses higher activity toward collagen and collagenlike sequences. The result of the thermal stability study indicated that PrtV was thermostable up to 40 C; at 50 C, stability gradually decreased. In addition, PrtV showed higher storage stability at -20 and 4 C, respectively, than at 25 C. Compared with collagenases from Clostridium histolyticum and Vibrio alginolyticus, PrtV was immunologically different and had no significant effect on the growth of CHO, HeLa, and Vero cells. Taken together, the results of the studies described in this paper advance our knowledge concerning the metal content and biochemical properties of PrtV.

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Section: Collagenasesmentioning

confidence: 99%

Metal Content and Biochemical Analyses of a Recombinant Collagenase PrtV from Vibrio parahaemolyticus

Yap

Lee

2000

Microbiology and Immunology

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“…Bacterial collagenase is known to be a metal protease, since the optimum pH is neutral to slightly alkaline and the activity is blocked by chelating agents such as EDTA or EGTA. 6) But the optimum pH of our purified enzyme was slightly acidic (pH 5.5-7.0), and unlike conventional collagenase the activity was not inhibited by EDTA or EGTA. Remarkably, the purified enzyme was inhibited by o-phenanthroline, a major chelating agent for Fe and Zn ions.…”

Section: Discussionmentioning

confidence: 70%

“…Several bacteria with collagenase activity are known: Clostridium sp, [1][2][3] Achromobacter sp, 4) Vibrio sp, [5][6][7] Pseudomonas sp, 8,9) Streptomyces sp, 10) Streptococcus sp, 11) Bacillus sp, 12) and Cytophaga sp. 13) Of these, the collagenase produced by Clostridium histolyticum is the most widely studied.…”

mentioning

confidence: 99%

Purification and Properties of a New Type of Protease Produced byMicrobacterium liquefaciens

Kanayama

Sakai

2005

Bioscience, Biotechnology, and Biochemistry

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A bacterium, identified as Microbacterium liquefaciens MIM-CG-9535-I, was isolated from a soil sample taken from the industrial site of a gelatin manufacturer. A new type of protease, which restrictively decomposes gelatin at one or two positions, was purified from the bacterial culture. The molecular mass of the purified enzyme was 21 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme specifically degraded the -chain of gelatin with a molecular weight of 100 kDa into two peptides of 60 kDa and 40 kDa. Native collagen was not a substrate for the enzyme.

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“…The mechanism of marine collagen degradation in marine nitrogen recycling is rather unclear because there are only several reports about marine collagenolytic enzyme-producing bacteria and collagenases (1)(2)(3)(4)(5). Collagenolytic proteases include metalloproteinases, serine proteases, and other proteases.…”

mentioning

confidence: 99%

Hydrolysis of Insoluble Collagen by Deseasin MCP-01 from Deep-sea Pseudoalteromonas sp. SM9913

Zhao¹,

Chen²,

Zhao

et al. 2008

Journal of Biological Chemistry

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Collagens are the most abundant proteins in marine animals and their degradation is important for the recycling of marine nitrogen. However, it is rather unclear how marine collagens are degraded because few marine collagenolytic proteases are studied in detail. Deseasins are a new type of multidomain subtilases. Here, the collagenolytic activity of deseasin MCP-01, the type example of deseasins, was studied. MCP-01 had broad substrate specificity to various type collagens from terrestrial and marine animals. It completely decomposed insoluble collagen into soluble peptides and amino acids, and was more prone to degrade marine collagen than terrestrial collagen. Thirtyseven cleavage sites of MCP-01 on bovine collagen chains were elucidated, showing the cleavage is various but specific. As the main extracellular cold-adapted protease from deepsea bacterium Pseudoalteromonas sp. SM9913, MCP-01 displayed high activity at low temperature and alkaline range. Our data also showed that the C-terminal polycystic kidney disease (PKD) domain of MCP-01 was able to bind insoluble collagen and facilitate the insoluble collagen digestion by MCP-01. Sitedirected mutagenesis demonstrated that Trp-36 of the PKD domain played a key role in its binding to insoluble collagen. It is the first time that the structure and function of a marine collagenolytic protease, deseasin MCP-01, has been studied in detail. Moreover, the PKD domain was experimentally proven to bind to insoluble protein for the first time. These results imply that MCP-01 would play an important role in the degradation of deep-sea sedimentary particulate organic nitrogen.Collagens, which distribute in skin, scale, bone, tendon, teeth, and blood vessels are the major protein constituents of the extracellular matrix and the most abundant proteins in all higher organisms including marine animals. It is an important fraction of marine organic nitrogen, and its degradation is important for marine nitrogen recycling.Because the tightly coiled triple helical collagen molecule assembles into water-insoluble fibers, collagens are resistant to most proteases except for a limited number of collagenolytic proteases. The mechanism of marine collagen degradation in marine nitrogen recycling is rather unclear because there are only several reports about marine collagenolytic enzyme-producing bacteria and collagenases (1-5). Collagenolytic proteases include metalloproteinases, serine proteases, and other proteases. Of all the collagenolytic proteases, mammalian collagenases, which are mammalian matrix metalloproteinases have been investigated in more detail than collagenolytic proteases from bacteria (6). Studies on the collagenolytic proteases from marine bacteria are rather minor and preliminary. Some marine collagenolytic enzyme-producing bacteria have been reported (1, 5). The collagenase from marine Vibro B-30 was purified and primarily characterized, which was reported at 1978 and 1980 (2-4). Since then, there has been no report on the characterization of marine bacterial col...

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Purification and characterization of a marine bacterial collagenase

Cited by 39 publications

References 28 publications

Metal Content and Biochemical Analyses of a Recombinant Collagenase PrtV from Vibrio parahaemolyticus

Metal Content and Biochemical Analyses of a Recombinant Collagenase PrtV from Vibrio parahaemolyticus

Purification and Properties of a New Type of Protease Produced byMicrobacterium liquefaciens

Hydrolysis of Insoluble Collagen by Deseasin MCP-01 from Deep-sea Pseudoalteromonas sp. SM9913

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