Premium
Fluorometric detection of interaction between lipase and glyceride
Author(s) -
Tsuzuki Wakako,
Kobayashi Shoichi,
Suzuki Tateo
Publication year - 1998
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1007/s11746-998-0261-y
Subject(s) - lipase , chemistry , glyceride , tryptophan , hydrolysis , residue (chemistry) , substrate (aquarium) , fluorescence , triacylglycerol lipase , chromatography , enzyme , organic chemistry , biochemistry , fatty acid , physics , amino acid , quantum mechanics , oceanography , geology
Abstract Previously, we devised the efficient modification of lipase, which can be dissolved and still maintain its activity in organic solvents. In this work, the fluorescence of the modified lipase could be detected in chloroform. When glycerides were added to the modified lipase solution, the intrinsic tryptophan fluorescence of the modified lipase decreased, which suggests that the environment of the tryptophan residue was affected by the substrate. The interaction between the modified lipase and glyceride was studied kinetically in terms of fluorescence intensity of the tryptophan residue. Because glyceride is not subject to hydrolysis in nonaqueous solution, the dissociation constant of the enzyme‐substrate complex could be determined. Thus, insight into the direct interaction between enzyme and substrate provided some structural information regarding the active site of lipase.