Elastase assays

Methods Enzymol. 1994;235:554-62. doi: 10.1016/0076-6879(94)35170-8.

Abstract

Two methods of P. aeruginosa elastase purification are described: Method 1 involves concentration of sample supernatants, followed by DEAE-Sepharose liquid chromatography, whereas Method 2 involves initial fractionations followed by molecular sieving and hydrophobic interaction high-performance liquid chromatography. The choice of methods depends on the available equipment and supplies. The methods of assaying elastase activity described as useful for a variety of applications. The elastin-nutrient agar plate method is a qualitative assay to determine the presence of elastase activity produced by a given culture or colony. Use of the quantitative elastin-Congo red assay is appropriate for determining elastase activities of mid-to-high elastase-producing cultures. For more sensitive determinations of P. aeruginosa elastase activity, use of the fluorogenic substrate is advisable.

Publication types

  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / analysis*
  • Bacterial Proteins / isolation & purification
  • Cattle
  • Chromatography, Agarose
  • Chromatography, High Pressure Liquid
  • Congo Red
  • Elastin / metabolism
  • Fluorescent Dyes
  • Molecular Sequence Data
  • Oligopeptides
  • Pancreatic Elastase / analysis*
  • Pancreatic Elastase / isolation & purification
  • Pseudomonas aeruginosa / enzymology*

Substances

  • Bacterial Proteins
  • Fluorescent Dyes
  • Oligopeptides
  • 2-aminobenzoylalanyl-glycyl-leucyl-alanyl-4-nitrobenzylamide
  • Congo Red
  • Elastin
  • Pancreatic Elastase