10.1002/cbic.201800734
ChemBioChem
FULL PAPER
(0.2-12 mM) were incubated in 50 mM HEPES, pH = 7.5. Aliquots (10 µL)
were removed at different time points (2 – 30 min after adding the malonate
derivative) and mixed with 90 µL of a 0.7 mM solution of DTNB. E412nm was
determined over a period of 5 min or until constant values were reached.
Linearity and stability of the reaction determining the remaining CoA were
proven using a calibration curve obtained from CoA dilutions. Initial
velocities in the linear range of conversion were determined by linear
regression. The velocity of three replicates was determined separately.
Michalis-Menten kinetics were calculated using OriginPro 2017G
(b9.4.0.220) using standard parameters and weighted values of the three
replicates.
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Part of this research was funded by the DFG Research Training
Group GRK 2341 “Microbial Substrate Conversion” (MiCon).
M. Grote is fellow of the International Max Planck Research
School of Chemical and Molecular Biology and thanks the
Studienstiftung des deutschen Volkes. This work is supported by
the „Center for Solvation Science ZEMOS“ funded by the German
Federal Ministry of Education and Research BMBF and by the
Ministry of Culture and Research of Nord Rhine-Westphalia.
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