Isolation by affinity chromatography of neutral proteinase from Clostridium histolyticum

Sparrow, Lindsay G.; McQuade, A.B.

doi:10.1016/0005-2744(73)90011-9

Cited by 23 publications

(10 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Vacuolating and lethal toxins were isolated and partially purified as previously described (Sparrow and McQuade 1973;Steward 1936). Six ATCC reference strains of C. histolyticum (ATCC no.…”

Section: Purification Of Lethal and Vacuolating Toxinmentioning

confidence: 99%

“…Culture broth filtrates of C. histolyticum contain several proteolytic enzymes, such as collagenase (Mandl et al 1957;Kono 1968;Mookhtiar and van Wart 1992;Finegold et al 2005), clostripain (Mitchell and Harrington 1968;Giles et al 1979;Ullmann and Jakubke 1994), elastase (Takasashi et al 1970), non-specific exopeptidases (Mandl et al 1957), proteinases (Sparrow and McQuade 1973;Kessler and Yaron 1973;Bicsak and Harper 1985), aminopeptidase (Kessler and Yaron 1973) and gelatinase (Yoshihara et al 1994). Furthermore, they have lethal and hemolytic activities (Steward 1936;Oakley and Warrack 1950).…”

Section: Introductionmentioning

confidence: 95%

See 1 more Smart Citation

Vacuolating, lethal, collagenase and clostripain activities of six reference ATCC Clostridium histolyticum strains

Jankowska‐Steifer

Martirosian

Ekiel

et al. 2010

World J Microbiol Biotechnol

View full text Add to dashboard Cite

We have previously demonstrated that C. histolyticum reference strain ATCC 19401 produces not only lethal factor but also hitherto unrecognized vacuolating toxin. The aim of this study was to compare vacuolating and lethal activities of six reference C. histolyticum strains (ATCC 6282, 8034, 17859, 17860, 19401 and 25770) and to determine whether production of vacuolating toxin is straindependent and how the amounts of both toxins produced by the same strain are related to each other and also to protease, collagenase and clostripain activities. All strains produced vacuolating and lethal toxins as well as collagenase, clostripain and proteases, but with different yield. Strain ATCC 19401 demonstrated considerable vacuolating and lethal activities and low activity of collagenase, clostripain and proteases. In other strains such relationship was not evident. Positive correlations were observed in collagenase and clostripain activities of all studied C. histolyticum strains (r = 0.71). Positive correlations were detected also in vacuolating activities of studied strains and clostripain (r = 0.62) and collagenase (r = 0.87) production, and this effect was statistically significant (P \ 0.05). Negative non-significant correlations were detected: (a) in lethal activities of studied strains and clostripain, or collagenase activities, (b) in vacuolating activities and protease production.

show abstract

“…Vacuolating and lethal toxins were isolated and partially purified as previously described (Sparrow and McQuade 1973;Steward 1936). Six ATCC reference strains of C. histolyticum (ATCC no.…”

Section: Purification Of Lethal and Vacuolating Toxinmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Vacuolating, lethal, collagenase and clostripain activities of six reference ATCC Clostridium histolyticum strains

Jankowska‐Steifer

Martirosian

Ekiel

et al. 2010

World J Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…DISCUSSION 5 ) reported that the neutral proteinase from Clostridium histolyticum was adsorbed on a column of Sepharoylcaproyl-glycyl-L-leucine (Sepharoyl-Cap-GlyLeu-OH), equilibrated with the starting buffer at pH 7.2, and eluted with 0.05 M Tris at pH 10.…”

Section: Effect Of Ph On Adsorption Of the Neutral Proteinase On The mentioning

confidence: 99%

Affinity Chromatography of Neutral Proteinase fromStreptomyces naraensison a Column ofN-Carbobenzoxy-l-phenylalanyl-aminohexyl-Sepharose

Hiramatsu

1982

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

“…C. histolyticum produces a large amount of collagenase class I (ColG) and collagenase class II (ColH), and the two collagenases decompose collagens in the ECM (Seifter and Harper 1971;Yoshihara et al 1994;Matsushita et al 1999). The microorganism also secretes a cysteine endopeptidase, clostripain (CP) (Ullmann and Bordusa 2004), and a metalloprotease, clostridial neutral protease (NP) (Sparrow and McQuade 1973). CP cleaves the carboxyl terminal side peptide bonds of Arg residues, like trypsin, from the pancreatic exocrine.…”

Section: Introductionmentioning

confidence: 99%

Cloning a neutral protease of Clostridium histolyticum, determining its substrate specificity, and designing a specific substrate

Maeda

Nakagawa

Murayama

et al. 2015

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Islet transplantation is a prospective treatment for restoring normoglycemia in patients with type 1 diabetes. Islet isolation from pancreases by decomposition with proteolytic enzymes is necessary for transplantation. Two collagenases, collagenase class I (ColG) and collagenase class II (ColH), from Clostridium histolyticum have been used for islet isolation. Neutral proteases have been added to the collagenases for human islet isolation. A neutral protease from C. histolyticum (NP) and thermolysin from Bacillus thermoproteolyicus has been used for the purpose. Thermolysin is an extensively studied enzyme, but NP is not well known. We therefore cloned the gene encoding NP and constructed a Bacillus subtilis overexpression strain. The expressed enzyme was purified, and its substrate specificity was examined. We observed that the substrate specificity of NP was higher than that of thermolysin, and that the protein digestion activities of NP, as determined by colorimetric methods, were lower than those of thermolysin. It seems that decomposition using NP does not negatively affect islets during islet preparation from pancreases. Furthermore, we designed a novel substrate that allows the measurement of NP activity specifically in the enzyme mixture for islet preparation and the culture broth of C. histolyticum. The activity of NP can also be monitored during islet isolation. We hope the purified enzyme and this specific substrate contribute to the optimization of islet isolation from pancreases and that it leads to the success of islet transplantation and the improvement of the quality of life (QOL) for diabetic patients.

show abstract

Isolation by affinity chromatography of neutral proteinase from Clostridium histolyticum

Cited by 23 publications

References 16 publications

Vacuolating, lethal, collagenase and clostripain activities of six reference ATCC Clostridium histolyticum strains

Vacuolating, lethal, collagenase and clostripain activities of six reference ATCC Clostridium histolyticum strains

Affinity Chromatography of Neutral Proteinase fromStreptomyces naraensison a Column ofN-Carbobenzoxy-l-phenylalanyl-aminohexyl-Sepharose

Cloning a neutral protease of Clostridium histolyticum, determining its substrate specificity, and designing a specific substrate

Contact Info

Product

Resources

About