The synthesis and analytical use of a highly sensitive and convenient substrate of elastase

https://doi.org/10.1016/0006-2944(74)90134-3Get rights and content

Abstract

This paper describes the synthesis and analytical use of succinyl-(l-alanine)3-p-nitroanilide, a new elastase substrate. At pH 8.0 and 25°C, the kinetic constants Km and kcat are 1.15 mm and 16.6 sec−1, respectively, in the absence of organic solvent and 2.4 mm and 21.2 sec−1 in the presence of 0.92% (vv) N-methylpyrrolidone. An expeditious and highly sensitive spectrophotometric method for measuring elastase activity is described: enzyme concentrations as low as 0.05 μg/ml may be assessed accurately within a few minutes. The procedure may be used to assay elastase in crude biological material since the substrate is virtually not attacked by trypsin or chymotrypsin and does not undergo appreciable spontaneous hydrolysis. The new substrate may also be used to detect elastase on polyacrylamide gels.

References (14)

  • L. Visser et al.

    Biochim. Biophys. Acta

    (1972)
  • J. Bieth et al.

    Anal. Biochem.

    (1973)
  • J. Bieth et al.

    Biochem. Biophys. Res. Commun.

    (1973)
  • G. Feinstein et al.

    Biochem. Biophys. Res. Commun.

    (1973)
  • B.F. Erlanger et al.

    Arch. Biochem. Biophys.

    (1961)
  • D.M. Shotton

    Methods Enzymol.

    (1970)
  • I.J. Schneider et al.

    Gastroenterology

    (1962)
There are more references available in the full text version of this article.

Cited by (0)

3

Chargé de Recherche à l'I.N.S.E.R.M.

View full text