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. 2023 Jan 27;15(2):109. doi: 10.3390/toxins15020109

Figure 5.

Figure 5

Kinetics parameters of the brown spider recombinant phospholipase-D (LiRecDT1). (A) Michaelis-Menten plot in the presence of different sphingomyelins (SM/LSM). (B) Michaelis-Menten plot in the presence of different LPCs and PCs. PLD activity was measured using the Amplex Red kit (Invitrogen). LiRecDT1 (1 µg) was added to different substrates (0.0024 to 5 mM), and the Amplex Red reagent mixture. After incubation, fluorescence was measured by fluorimeter, excitation 540 nm, and emission 570 nm (Tecan Infinite M200). The experiments were performed in 100 mM Tris-HCl pH: 7.4, 100 mM NaCl, and 5 mM MgCl2, and 0.01% Triton X-100 at 37 °C. The parameters were calculated using Prism 6.0 and shown as the mean value.