Kinetic and structural characterization of fumonisin esterases

Published: 2 October 2024| Version 1 | DOI: 10.17632/3vmzmb9k6y.1
Contributor:
Daniel Janos Incze

Description

Raw data for the kinetic characterization of two fumonisin esterases. The dataset belongs to the paper "(the citation will be updated when the paper is published)" by Incze et al., and the research is described there in details. In short, all substrates and enzyme solutions were diluted in Buffer A (50 mM potassium phosphate buffer supplemented with 100 μg/mL of BSA, at pH 6.0) for the enzyme kinetic measurements. The fermentation supernatants containing FE were diluted to a final enzyme concentration of 2.0 μg/L in the reactions. Solutions of FB1 and pHFB1_7 were diluted to five different concentrations in the range between 4.5–165 μM in final reaction volumes of 200 μL. The reaction mixtures were incubated at 37 °C in a TDB-120 Dry Block Thermostat (Biosan, Riga, Latvia). Samples (20 μL) were taken from every enzyme reaction, five times at different time points, depending on the substrate, substrate concentration, and enzyme. The samples were mixed immediately with 113 μL of MeOH to terminate theenzymatic reaction. The reactions were performed in triplicates and were analyzed with HPLC-FLD. Linear equations were fitted to the measured points of the product concentration-time function’s diagram with linear regression using Microsoft Excel. During the degradation of FB1, only those measurement points where the amount of HFB1 was negligible were evaluated for the fitting. The initial reaction rates were defined as the slope of the linear equation in μM/min. The fitted linear equations were accepted when the R^2 values were at least 0.995. The Excel data file consists of 4 sheets. Sheets named "FE1, FB1", "FE1, pHFB1_7", "FE2, FB1" and "FE2, pHFB1_7" correspond to the datasets of FB1 hydrolysis by FE1, pHFB1_7 hydrolysis by FE1, FB1 hydrolysis by FE2 and pHFB1_7 hydrolysis by FE2, respectively. Columns "c initial(FB1) [µM]", "c initial(pHFB1_7) [µM]", "t [min]", "c (FB1) [µM]", "c (pHFB1_7) [µM]", and c (HFB1) [µM] contains the data of the concentration of FB1 in µM prior enzymatic reaction, the concentration of pHFB1_7 in µM prior enzymatic reaction, reaction time in minutes, actual concentration of FB1 in µM, actual concentration of pHFB1_7 in µM and actual concentration of HFB1 in µM, respectively.

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Categories

Biocatalysis Kinetics

Funding

Nemzeti Kutatási Fejlesztési és Innovációs Hivatal

TKP2021-EGA-02

Nemzeti Kutatási Fejlesztési és Innovációs Hivatal

C1341189

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