1887

Abstract

The number and approximate molecular weights of extracellular alkaline proteases produced by were determined by gelatin-PAGE. Three major bands of protease activity with apparent molecular weights of approximately 28000, 22500 and 19500 (proteases 1, 2 and 3, respectively) and two minor bands of protease activity with apparent molecular weights of approximately 15500 and 14500 (proteases 4 and 5, respectively) were obtained after gelatin-PAGE. The activities of the five proteases were inhibited by serine protease inhibitors but their activities were not affected by inhibitors of trypsin-like enzymes. Histidine, which inhibited collagenase, did not inhibit the activities of the alkaline serine proteases. The production of protease 1, however, was enhanced by histidine. Protease 1 production was also affected by temperature and production was depressed at 37 °C. Gelatin-PAGE of a commercial collagenase preparation revealed four bands of activity which were identified as collagenases with apparent molecular weights of approximately 45000, 38500, 33500 and 31000. The collagenase preparation was contaminated with two serine proteases. The release of [H]proline from collagen matrices produced by smooth muscle cells was shown to be a sensitive assay for bacterial collagenases and was used to show that produced a basal constitutive level of extracellular collagenase. The constitutive levels of collagenase were affected by aeration.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-4-1141
1983-04-01
2024-12-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/4/mic-129-4-1141.html?itemId=/content/journal/micro/10.1099/00221287-129-4-1141&mimeType=html&fmt=ahah

References

  1. Hare P., Long S., Robb F. T., Woods D. R. 1981; Regulation of exoprotease production by temperature and oxygen inVibrio alginolyticus. Archives of Microbiology 130:276–280
    [Google Scholar]
  2. Heussen C., Dowdle E. B. 1980; Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulphate and copolymerized substrates. Analytical Biochemistry 102:196–202
    [Google Scholar]
  3. Jones P. A., Scott-Burden T. 1979; Activated macrophages digest the extracellular matrix proteins produced by cultured cells. Biochemical and Biophysical Research Communications 86:71–77
    [Google Scholar]
  4. Keil-Dlouha V. 1976; Chemical characterization and study of the autodigestion of pure collagenase fromAchromobacter iophagus. Biochimica et biophysica acta 429:239–251
    [Google Scholar]
  5. Keil-Dlouha V., Keil B. 1978; Subunit structure ofAchromobacter collagenase. Biochimica et biophysica acta 522:218–228
    [Google Scholar]
  6. Keil-Dlouha V., Misrahi R., Keil B. 1976; The induction of collagenase and a neutral proteinase by their high molecular weight substrates inAchromobacter iophagus. Journal of Molecular Biology 107:293–305
    [Google Scholar]
  7. Lecroisey A., Keil-Dlouha V., Woods D. R., Perrin D., Keil B. 1975; Purification, stability attva uuuuHiuw ui vuiiagdiaov iiuin ZILnrurnu-bacter iophagus. FEBS Letters 59:167–172
    [Google Scholar]
  8. Long S., Mothibeli M. A., Robb F. T., Woods D. R. 1981; Regulation of extracellular alkaline protease activity by histidine in a collagenolyticVibrio alginolyticus strain. Journal of General Microbiology 127:193–199
    [Google Scholar]
  9. Reid G. C., Robb F. T., Woods D. R. 1978; Regulation of extracellular collagenase production inAchromobacter iophagus. Journal of General Microbiology 109:149–154
    [Google Scholar]
  10. Reid G. C., Woods D. R., Robb F. T. 1980; Peptone induction and rifampicin insensitive collagenase production byVibrio alginolyticus. Journal of Bacteriology 142:447–454
    [Google Scholar]
  11. Robbertse P. J., Woods D. R., Reay A. M., Robb F. T. 1978; Simple and sensitive procedure for screening collagenolytic bacteria and the isolation of collagenase mutants. Journal of General Microbiology 106:373–376
    [Google Scholar]
  12. Welton R. L., Woods D. R. 1973; Halotolerant collagenolytic activity ofAchromobacter iophagus. Journal of General Microbiology 75:191–196
    [Google Scholar]
  13. Wünsch E., Heidrich H. G. 1963; Zur quantitativen Bestimmung der Kollagenase. Hoppe-Seyler's Zeitschrift für physiologische Chemie 333:149–151
    [Google Scholar]
/content/journal/micro/10.1099/00221287-129-4-1141
Loading
/content/journal/micro/10.1099/00221287-129-4-1141
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error