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A sensitive trimethylamine‐ N ‐oxide aldolase assay in two steps without deproteinisation
Author(s) -
Nielsen Michael K,
Havemeister Wiebke,
Rehbein Hartmut,
Sotelo Carmen G,
Jørgensen Bo M
Publication year - 2000
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/(sici)1097-0010(20000115)80:2<197::aid-jsfa513>3.0.co;2-u
Subject(s) - aldolase a , trimethylamine , chemistry , trimethylamine n oxide , enzyme , chromatography , oxide , enzyme assay , ph indicator , biochemistry , combinatorial chemistry , organic chemistry
A two‐step assay for trimethylamine‐ N ‐oxide aldolase (the target enzyme) is described in which the second step, the indicator reaction, is of the enzymatic end‐point type. This indicator reaction is carried out at a slightly alkaline pH, outside the target enzyme's active pH range. An effective inactivation of the target enzyme is thus obtained solely by shifting the pH to that of the indicator reaction, thereby avoiding a deproteinisation step. By inserting a downward pH step as the stopping method, many samples may be collected and applied to the indicator reaction simultaneously. These features make the method very productive and easy to perform. © 2000 Society of Chemical Industry