A quantitative assay for agarase activity determination using agarose-iodine complex

Anal Biochem. 2022 Mar 15:641:114560. doi: 10.1016/j.ab.2022.114560. Epub 2022 Jan 20.

Abstract

Rapid and simple spectrophotometric methods are required to detect various oligosaccharides produced by agar-hydrolysing enzymes. Herein, we present a quantitative agarose-iodine assay for agarase activity determination via the detection of the extent of agarose degradation. The agarose-iodine complex becomes reddish orange upon the addition of Lugol solution, and the enzymatic activity can be detected with ultraviolet-visible spectroscopy at 600 nm. The main advantages of this modified Lugol assay are high sensitivity, simple detection, and cost effectiveness. A novel definition of the unit to measure and compare the activities of agarases is also suggested.

Keywords: 3,5-Dinitrosalicylic acid assay; Agarase; Agarose–iodine complex; Lugol solution; Nelson–Somogyi assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alteromonadaceae / enzymology
  • Fluorescent Dyes / chemistry
  • Glycoside Hydrolases / analysis*
  • Glycoside Hydrolases / metabolism
  • Iodides / chemistry
  • Iodine / chemistry*
  • Sepharose / chemistry*
  • Spectrophotometry, Ultraviolet

Substances

  • Fluorescent Dyes
  • Iodides
  • Sepharose
  • Iodine
  • Glycoside Hydrolases
  • agarase
  • Lugol's solution

Supplementary concepts

  • Agarivorans albus