TY  - JOUR
A1  - Kagel, Heike
A1  - Bier, Frank Fabian
A1  - Frohme, Marcus
A1  - Glökler, Jörn F.
T1  - A Novel Optical Method To Reversibly Control Enzymatic Activity Based On Photoacids
JF  - Scientific reports
N2  - Most biochemical reactions depend on the pH value of the aqueous environment and some are strongly favoured to occur in an acidic environment. A non-invasive control of pH to tightly regulate such reactions with defined start and end points is a highly desirable feature in certain applications, but has proven difficult to achieve so far. We report a novel optical approach to reversibly control a typical biochemical reaction by changing the pH and using acid phosphatase as a model enzyme. The reversible photoacid G-acid functions as a proton donor, changing the pH rapidly and reversibly by using high power UV LEDs as an illumination source in our experimental setup. The reaction can be tightly controlled by simply switching the light on and off and should be applicable to a wide range of other enzymatic reactions, thus enabling miniaturization and parallelization through non-invasive optical means.
Y1  - 2019
U6  - https://doi.org/10.1038/s41598-019-50867-w
SN  - 2045-2322
VL  - 9
PB  - Nature Publishing Group
CY  - London
ER  - 
TY  - JOUR
A1  - Fischbach, Jens
A1  - Loh, Qiuting
A1  - Bier, Frank Fabian
A1  - Lim, Theam Soon
A1  - Frohme, Marcus
A1  - Glökler, Jörn
T1  - Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method
JF  - Scientific reports
N2  - We identified Alizarin Red S and other well known fluorescent dyes useful for the online detection of pyrophosphate in enzymatic assays, including the loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. An iterative screening was used for a selected set of compounds to first secure enzyme compatibility, evaluate inorganic pyrophosphate sensitivity in the presence of manganese as quencher and optimize conditions for an online detection. Of the selected dyes, the inexpensive alizarin red S was found to selectively detect pyrophosphate under LAMP and PCR conditions and is superior with respect to its defined red-shifted spectrum, long shelf life and low toxicity. In addition, the newly identified properties may also be useful in other enzymatic assays which do not generate nucleic acids but are based on inorganic pyrophosphate. Finally, we propose that our screening method may provide a blueprint for rapid screening of compounds for detecting inorganic pyrophosphate.
Y1  - 2017
U6  - https://doi.org/10.1038/srep45085
SN  - 2045-2322
VL  - 7
PB  - Nature Publ. Group
CY  - London
ER  -