Applied Biochemistry and Biotechnology

, Volume 165, Issue 1, pp 69–74

Reversible Inhibition of Esterase Activity After Separation and Immobilization

Authors

  • Takahiro Sakikawa
    • Graduate School of Science and Engineering (Science Section)Ehime University
    • Venture Business LaboratoryEhime University
    • Graduate School of Science and Engineering (Science Section)Ehime University
    • Venture Business LaboratoryEhime University
Article

DOI: 10.1007/s12010-011-9233-z

Cite this article as:
Sakikawa, T. & Shimazaki, Y. Appl Biochem Biotechnol (2011) 165: 69. doi:10.1007/s12010-011-9233-z

Abstract

An inhibitor, 9-amino-1,2,3,4-tetra hydroacridine (tacrine), is a reversible inhibitor of esterases. The reversible inhibition of the enzyme activity is thought to be examined after separation and immobilization of the enzyme under non-denaturing conditions. Hydrolytic changes of phosphatidylcholine by carboxylesterase were obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after the esterase was separated by non-denaturing two-dimensional electrophoresis, was immobilized to membranes and was stained by Ponceau S. The changes were inhibited after the enzyme on the membrane was treated by tacrine. Furthermore, the hydrolytic activity of the esterase was recovered after the inhibitor was washed with aspartic acid solution. These results indicate that the phosphatidylcholine hydrolysis activity of the isolated and immobilized enzyme is reversibly inhibited under non-denaturing conditions. Furthermore, this method can be developed to the production of an enzyme reactor able to regulate amounts of lipids.

Keywords

ElectrophoresisPonceau SMALDI-TOF MSTacrinePhosphatidylcholine

Abbreviations

2-DE

Two-dimensional electrophoresis

PVDF

Polyvinylidene fluoride

Tacrine

9-Amino-1,2,3,4-tetra hydroacridine

MALDI-TOF MS

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

CBB

Coomassie brilliant blue

Copyright information

© Springer Science+Business Media, LLC 2011