Drug metabolite synthesis by immobilized human FMO3 and whole cell catalysts

Abstract

Background Sufficient reference standards of drug metabolites are required in the drug discovery and development process. However, such drug standards are often expensive or not commercially available. Chemical synthesis of drug metabolite is often difficulty due to the highly regio- and stereo-chemically demanding. The present work aims to construct stable and efficient biocatalysts for the generation of drug metabolites in vitro. Result In this work, using benzydamine as a model drug, two easy-to-perform approaches (whole cell catalysis and enzyme immobilization) were investigated for the synthesis of FMO3-generated drug metabolites. The whole cell catalysis was carried out by using cell suspensions of E. coli JM109 harboring FMO3 and E. coli BL21 harboring GDH (glucose dehydrogenase), giving 1.2 g/L benzydamine N -oxide within 9 h under the optimized conditions. While for another approach, two HisTrap HP columns respectively carrying His 6 -GDH and His 6 -FMO3 were connected in series used for the biocatalysis. In this case, 0.47 g/L benzydamine N -oxide was generated within 2.5 h under the optimized conditions. In addition, FMO3 immobilization at the C-terminal (membrane anchor region) significantly improved its enzymatic thermostability by more than 10 times. Moreover, the high efficiency of these two biocatalytic approaches was also confirmed by the N -oxidation of tamoxifen. Conclusions The results presented in this work provides new possibilities for the drug-metabolizing enzymes-mediated biocatalysis. Electronic supplementary material The online version of this article (10.1186/s12934-019-1189-7) contains supplementary material, which is available to authorized users.

Bioconversion of vitamin D3 to bioactive calcifediol and calcitriol as high-value compounds

“…In general, enzymes and whole cells are two categories of biocatalysts commonly used for bioconversion [18,19]. Here, we review advances in the characterization and engineering of VD 3 hydroxylase that plays a key role in VD 3 bioconversion, and progress in whole-cell bioconversion, which can use inexpensive and abundant raw materials and avoiding the addition of expensive cofactors.…”

Immobilised-enzyme microreactors for the identification and synthesis of conjugated drug metabolites

“…21 There are also some reports of the application of ow-through reactors to cytochromes P450 activity, however these are limited in scope and focus on the prediction of metabolism, rather than the generation of metabolites. [18][19][20][21][22][23][24][25] Microuidic ow synthesis platforms have demonstrated the ability to bypass challenges around batch synthesis. Enzyme immobilisation has been widely utilised throughout the literature.…”

Expression of Concern to: Drug metabolite synthesis by immobilized human FMO3 and whole cell catalysts

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The Editor-in-Chief is issuing an Editorial Expression of Concern as after publication of this article [1], it has come to light that the biological materials were used and the experiments were conducted without proper authorisation from the laboratory where these data were obtained. In addition, at the time when the research was performed, the first author was a PhD student in the Department of Life Sciences and Systems Biology, University of Torino, and the second author was a master's student in the Department of Culture, Politics and Society, University of Torino.

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