Bioluminescence Inhibition of Bacterial Luciferase by Aliphatic Alcohol, Amine and Carboxylic Acid: Inhibition Potency and Mechanism

Abstract

The inhibitory effects of hydrophobic molecules on the bacterial luciferase, BL, luminescence reaction were analyzed using an electrochemically-controlled BL luminescence system. The inhibition potency of alkyl amines, CnNH2, and fatty acids, CmCOOH (m = n -1), on the BL reaction increased with an increase in the alkyl chain-length of these aliphatic compounds. CmCOOH showed lower inhibition potency than CnNH2 and alkyl alcohols, CnOH, data for which have been previously reported. To make clear the inhibition mechanisms of the aliphatic compounds on the BL reaction, the initial rate of the BL reaction was measured and analyzed using the Dixon plot and Cornish-Bowden plot. The C12OH inhibited the BL reaction in competition with the substrate C11CHO, while C12NH2 and C11COOH inhibited in an uncompetitive manner with the C11CHO. These results suggest that the alkyl chain-length and the terminal unit of the aliphatic compound determine the inhibition potency and the inhibition mechanism, respectively.

Electrochemical Control of Bioluminescence by Blocking the Adsorption of the Bacterial Luciferase Using a Mercaptobipyridine Self-assembled Monolayer

“…[1][2][3][4][5][6] The effect of hydrophobic molecules, such as terminally-substituted normal alkyl compounds, on the bacterial luciferase (BL) bioluminescence has been reported in our previous studies. 5,6 The BL reaction requires a reduced flavin mononucleotide (FMNH2) as one of the substrates. 7 However, the FMNH2 is rapidly oxidized to oxidized flavin (FMN) by dissolved oxygen in an aqueous solution.…”

AgroLux: bioluminescent Agrobacterium to improve molecular pharming and study plant immunity

“…A key advantage of bioluminescence‐based imaging is the capability to monitor only living bacteria, as light emission depends on the cofactors ATP, NADPH and the reduced form of flavin mononucleotide (FMNH 2 ) (Gregor et al ., 2018). Light emission is determined by many factors, including bacterial quantity, the expression of the lux operon, cofactor availability, the stability of luciferase, and the absence of quenchers and luciferase inhibitors (Gregor et al ., 2018; Yamasaki et al ., 2013). However, in biosensors, luciferase levels are relatively low, and light is often directly correlated with transcript levels.…”