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

Area of Science:

• Biochemistry
• Enzymology
• Spectroscopy

Background:

• Agar-hydrolyzing enzymes produce oligosaccharides, necessitating rapid detection methods.
• Existing spectrophotometric assays for agarase activity can be complex or lack sensitivity.

Purpose of the Study:

• To develop a simple, sensitive, and cost-effective spectrophotometric assay for quantifying agarase activity.
• To determine the extent of agarose degradation by agarase enzymes.

Main Methods:

• A modified Lugol assay using the agarose-iodine complex.
• Quantification of agarose degradation via ultraviolet-visible spectroscopy at 600 nm.

Main Results:

• The agarose-iodine complex shows a distinct reddish-orange color change upon agarose degradation.
• The assay demonstrates high sensitivity and simplicity for detecting agarase activity.

Conclusions:

• The developed agarose-iodine assay is a viable method for determining agarase activity.
• This assay offers advantages in sensitivity, simplicity, and cost-effectiveness for oligosaccharide detection.