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Issue 4, 2021
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An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C

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Abstract

The Bacillus cereus phosphatidylcholine-specific phospholipase C (PC-PLCBc) is an enzyme that catalyses the hydrolysis of phosphatidylcholines into phosphocholine and 1,2-diacylglycerols. PC-PLCBc has found applications in both the food industry and in medicinal chemistry. Herein, we report our work in the development and optimisation of a matrix assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry-based assay to monitor PC-PLCBc activity. The use of one-phase and two-phase reaction systems to assess the inhibition of PC-PLCBc with different structural classes of inhibitors was compared. We also highlighted the advantage of our assay over the commonly used commercially available Amplex Red assay. This method will also be applicable to work on the activity and inhibition of other phospholipases.

Graphical abstract: An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C

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Supplementary files

Article information


Submitted
02 Dec 2020
Accepted
29 Dec 2020
First published
29 Dec 2020

Anal. Methods, 2021,13, 491-496
Article type
Paper

An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C

N. Sharma, R. J. Langley, C. Eurtivong, E. Leung, R. J. Dixon, E. K. Paulin, S. W. P. Rees, L. I. Pilkington, D. Barker, J. Reynisson and I. K. H. Leung, Anal. Methods, 2021, 13, 491
DOI: 10.1039/D0AY02208J

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