Production of an O-desmethylated product, a major human metabolite, of rabeprazole sulfide by bacterial P450 enzymes

• Published in: Enzyme and microbial technology 2023-12, Vol.171, p.110328-110328, Article 110328
• Main Authors: Cao, Ngoc Tan, Cha, Gun Su, Kim, Jeong-Hoon, Lee, Yujin, Yun, Chul-Ho, Nguyen, Ngoc Anh
• Format: Article
• Language: English
• Subjects: CYP102A1 mutant; Cytochrome P450; O-desmethylation; Proton pump inhibitors; Rabeprazole sulfide
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2023.110328

Rabeprazole is a common type of proton pump inhibitor (PPI) used to treat various peptic disorders. Unlike most PPI drugs, rabeprazole is spontaneously reduced to rabeprazole sulfide (thioether) when it is given to patients. As a result, rabeprazole sulfide is considered one of the active metabolites of rabeprazole. Rabeprazole sulfide is mainly metabolized to desmethyl rabeprazole sulfide by CYP2C19 and CYP2D6 in people. However, the pharmacological efficacy and safety of desmethyl rabeprazole sulfide have not yet been investigated. Its usage is challenging due to the high cost associated with the drug. In this study, we found CYP102A1 mutants that can produce desmethyl rabeprazole sulfide as a major metabolite of rabeprazole sulfide. The chemical characteristics of the major product were confirmed using high-performance liquid chromatography, LC-mass spectrometry, and nuclear magnetic resonance spectroscopy. CYP102A1 mutants R47L/F87V/L188Q, R47L/F87V/L188Q/A335V/Q359R, and R47L/F87V/L188Q/I254V/D351E showed kcat values of 39, 93, and 88 min−1, respectively, for O-desmethylation of rabeprazole sulfide. Furthermore, the highest concentration of desmethyl rabeprazole sulfide product from 2 mM rabeprazole sulfide at optimal conditions was obtained in bacterial whole-cell biotransformation with the R47L/F87V/L188Q mutant, reaching 0.63 mM at 4-h incubation. In conclusion, we present a platform that facilitates the efficient and sustainable production of the desmethylated product from rabeprazole sulfide for use in the biopharmaceutical industry. [Display omitted] •A set of CYP102A1 mutants catalyze O-desmethylation of rabeprazole sulfide.•Major product of rabeprazole sulfide is desmethyl rabeprazole sulfide.•The major metabolite was characterized by LC-MS and NMR analyses.•A high active mutant showed 93 min-1 for desmethyl rabeprazole sulfide production.•Bacterial whole-cell system with a CYP102A1 mutant reached 0.63 mM product.