Applied Microbiology and Biotechnology

, Volume 29, Issue 5, pp 494–496 | Cite as

A note on the use of dinitrosalicylic acid for determining the products of enzymatic reactions

  • Michael J. Bailey
Applied Microbiology

Summary

Factors contributing to non-linearity of enzyme assays incorporating detection of reaction products using dinitrosalicylic acid (DNS) are discussed. The common practice of diluting reaction products before quantification of reducing compounds is shown to be one cause of non-linearity. Insufficiency of substrate is also an important contributory factor in most modern versions of the method, although the original procedure was correctly designed to ensure a linear reaction. The inherent inaccuracy involved in expression of the results of non-linear reactions in units implying linearity (katals or International Units) is emphasized.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bailey MJ (1981) The effect of β-glucosidase on some assays for cellulolytic enzymes. Biotechnol Lett 3:695–700Google Scholar
  2. Bailey MJ, Nevalainen KMH (1981) Induction, isolation and testing of stable Trichoderma reesei mutants with improved production of solubilizing cellulase. Enzyme Microb Technol 3:153–157Google Scholar
  3. Fisher EH, Stein (1961) α-Amylase from human saliva. Biochem Prep 8:27–33Google Scholar
  4. IUPAC (International Union of Pure and Applied Chemistry) (1987) Measurement of cellulase activities. Pure and Appl Chem 59:257–268Google Scholar
  5. Mandels M, Andreotti R, Roche C (1976) Measurement of saccharifying cellulase. Biotechnol Bioeng Symp 6:21–33Google Scholar
  6. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428Google Scholar
  7. Miller GL, Blum R, Glennon WE, Burton AL (1960) Measurement of carboxymethylcellulase activity. Anal Biochem 2:127–132Google Scholar
  8. Noelting G, Bernfeld P (1948) Sur les enzymes amylolytiques III. La β-amylase: dosage d'activité et controle de l'absence d'α-amylase. Helv Chim Acta 31:286–290Google Scholar
  9. Sumner JB (1921) Dinitrosalicylic acid: A reagent for the estimation of sugar in normal and diabetic urine. J Biol Chem 47:5–9Google Scholar
  10. Sumner JB (1925) A more specific reagent for the determination of sugar in urine. J Biol Chem 65:51–54Google Scholar
  11. Sumner JB, Howell SF (1935) A method for determination of saccharase activity. J Biol Chem 108:51–54Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Michael J. Bailey
    • 1
  1. 1.VTT, Biotechnical LaboratoryEspooFinland

Personalised recommendations