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Improving the Time Resolution for Remote Control of Enzyme Activity by a Nanosecond Laser‐Induced pH Jump
Author(s) -
Kohse Stefanie,
Neubauer Antje,
Lochbrunner Stefan,
Kragl Udo
Publication year - 2014
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201402442
Subject(s) - chemistry , enzyme , nanosecond , substrate (aquarium) , excitation , kinetics , laser , temporal resolution , resolution (logic) , pulse (music) , jump , biophysics , photochemistry , biochemistry , optics , physics , oceanography , quantum mechanics , artificial intelligence , detector , computer science , biology , geology
Application of light as a trigger for remote control of enzyme activation offers high temporal and spatial resolution. A known enzymatic system based on an acid phosphatase and 6‐chloro‐8‐fluoro‐4‐methylumbelliferone phosphate as substrate is employed to study different strategies for improving the time resolution of enzyme activation by a light‐induced pH jump. Hence, a single laser pulse excitation with a pulse duration of 6 ns resulted in a 4‐fold enzyme activation by instantaneous proton release through the rearrangement of 2‐nitrobenzaldehyde. The time resolution of the activation experiment is now limited only by the detection system, no longer by the excitation step. In addition, halogenated 2‐nitrobenzaldehydes are tested for their suitability as phototriggers and a detailed study of the pH‐dependent kinetics of the enzymatic reaction is performed, which reveals that the substrate can probably only bind to the enzyme in its monoanionic form.