Measuring Thiaminase Activity in Fish Extracts using Fluorescence Spectrophotometry.

ABSTRACT: Thiaminase is an enzyme that destroys thiamine (vitamin B1) and is present in various fishes, mussels, plants, and bacteria. A sensitive and versatile assay is needed to measure thiaminase activity in complex biological samples using common reagents and variable reaction conditions. We developed a simple assay that uses fluorescence spectrophotometry and a microplate reader to measure thiaminase activity in fish tissue extracts via the destruction of added thiamine over time. Thiamine concentration, cosubstrate choice and concentration, and pH can be varied according to the needs of the investigator. Using this assay, we successfully measured common enzyme kinetic constants for thiaminase from a Pacific herring extract. The enzyme exhibited Michaelis-Menten kinetics (V _max

SUBMITTER: Porter D 

PROVIDER: S-EPMC12715735 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature