Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1983 Jan;45(1):200–204. doi: 10.1128/aem.45.1.200-204.1983

Dual-Substrate Plate Diffusion Assay for Proteases

Thomas J Montville 1
PMCID: PMC242253  PMID: 6337548

Abstract

A plate diffusion assay for endopeptidases was developed. Proteases applied to plates containing 1% casein, 1% gelatin, and 1.5% agar caused distinct zones reminiscent of immunoprecipitation bands. The diameter of the zones was linearly proportional to the log of the enzyme activity applied over a range from 0.01 to greater than 100 IU/ml.

Full text

PDF
204

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Awerbuch T. E., Stark A. A. Plate diffusion assay as a rapid method for dosimetry of mutagens. Appl Environ Microbiol. 1979 Dec;38(6):1127–1131. doi: 10.1128/aem.38.6.1127-1131.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dasgupta B. R., Sugiyama H. Isolation and characterization of a protease from Clostridium botulinum type B. Biochim Biophys Acta. 1972 Jun 16;268(3):719–729. doi: 10.1016/0005-2744(72)90276-8. [DOI] [PubMed] [Google Scholar]
  3. ERLANGER B. F., KOKOWSKY N., COHEN W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys. 1961 Nov;95:271–278. doi: 10.1016/0003-9861(61)90145-x. [DOI] [PubMed] [Google Scholar]
  4. Kamekura M., Onishi H. Protease formation by a moderately halophilic Bacillus strain. Appl Microbiol. 1974 Apr;27(4):809–810. doi: 10.1128/am.27.4.809-810.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kiyohara H., Nagao K., Yana K. Rapid screen for bacteria degrading water-insoluble, solid hydrocarbons on agar plates. Appl Environ Microbiol. 1982 Feb;43(2):454–457. doi: 10.1128/aem.43.2.454-457.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. MCDONALD C. E., CHEN L. L. THE LOWRY MODIFICATION OF THE FOLIN REAGENT FOR DETERMINATION OF PROTEINASE ACTIVITY. Anal Biochem. 1965 Jan;10:175–177. doi: 10.1016/0003-2697(65)90255-1. [DOI] [PubMed] [Google Scholar]
  7. Montville T. J. Effect of plating medium on heat activation requirement of Clostridium botulinum spores. Appl Environ Microbiol. 1981 Oct;42(4):734–736. doi: 10.1128/aem.42.4.734-736.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Schwabe C. A fluorescent assay for proteolytic enzymes. Anal Biochem. 1973 Jun;53(2):484–490. doi: 10.1016/0003-2697(73)90098-5. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES