Abstract
Objectives
To find a simple enzymatic strategy for the efficient synthesis of the expensive 5′-hydroxyomeprazole sulfide, a recently identified minor human metabolite, from omeprazole sulfide, which is an inexpensive substrate.
Results
The practical synthetic strategy for the 5′-OH omeprazole sulfide was accomplished with a set of highly active CYP102A1 mutants, which were obtained by blue colony screening from CYP102A1 libraries with a high conversion yield. The mutant and even the wild-type enzyme of CYP102A1 catalyzed the high regioselective (98 %) C-H hydroxylation of omeprazole sulfide to 5′-OH omeprazole sulfide with a high conversion yield (85–90 %).
Conclusions
A highly efficient synthesis of 5′-OH omeprazole sulfide was developed using CYP102A1 from Bacillus megaterium as a biocatalyst.
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Acknowledgments
This research was supported by the National Research Foundation of Korea (Grant NRF-2016R1A2B4006978) and by the Next-Generation BioGreen 21 program (SSAC Grant# PJ011058), Rural Development Administration, Republic of Korea.
Supporting information
Supplementary Methods-Generation of the CYP102A1 libraries.
Supplementary Methods-Screening of the CYP102A1 libraries.
Supplementary Methods-Liquid chromatography-mass spectrometry analysis.
Supplementary Methods-Identification of the omeprazole sulfide metabolite by NMR spectroscopy.
Supplementary Table 1. Mutated amino acid residues of the CYP102A1 mutants used in this study.
Supplementary Table 2. 1H chemical shifts for 5'-OH omeprazole sulfide.
Supplementary Figure 1. Metabolic pathway of omeprazole in humans.
Supplementary Figure 2. Screening of CYP102A1 mutant libraries.
Supplementary Figure 3. Close look at the 3.81 ppm and NOE results of 5′-OH omeprazole sulfide.
Supplementary Figure 4. The mutation sites for CYP102A1 M16V2 and its mutants.
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Jang, HH., Ryu, SH., Le, TK. et al. Regioselective C-H hydroxylation of omeprazole sulfide by Bacillus megaterium CYP102A1 to produce a human metabolite. Biotechnol Lett 39, 105–112 (2017). https://doi.org/10.1007/s10529-016-2211-3
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DOI: https://doi.org/10.1007/s10529-016-2211-3