Alternative method for quantification of alfa-amylase activity

Braz J Biol. 2010 May;70(2):405-7. doi: 10.1590/s1519-69842010005000008. Epub 2010 Mar 19.

Abstract

A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 A degrees C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing alpha-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale alpha-amylase over a wide range (2.4 to 7,500 U.mL-1) in scientific investigation as well as in teaching laboratory activities.

MeSH terms

  • Agar
  • Aspergillus oryzae / enzymology*
  • Chemistry Techniques, Analytical / economics
  • Chemistry Techniques, Analytical / methods*
  • Diffusion
  • Reproducibility of Results
  • alpha-Amylases / analysis*

Substances

  • Agar
  • alpha-Amylases