An assay for thiaminase I in complex biological samples.
An alternative method for measuring thiaminase I activity in complex samples is described. This assay is based on the selective consumption of the highly chromophoric 4-nitrothiophenolate by thiaminase I, resulting in a large decrease in absorbance at 411nm. This new assay is simple and sensitive, and it requires only readily available chemicals and a visible region spectrophotometer. In addition, the assay is optimized for high-throughput analysis in a 96-well format with complex biological samples.
Hanes
Jeremiah W
JW
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA.
Kraft
Clifford E
CE
Begley
Tadhg P
TP
D019883:Alkyl and Aryl Transferases / Q000032:analysis* / Q000378:metabolism; D045424:Complex Mixtures / Q000737:chemistry; D006867:Hydrolases / Q000032:analysis / Q000378:metabolism; D007700:Kinetics; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism
D045424:Complex Mixtures; D013438:Sulfhydryl Compounds; C014712:4-nitrothiophenolate; D019883:Alkyl and Aryl Transferases; C022737:thiamin pyridinylase; D006867:Hydrolases; C022731:thiaminase II