Spray-Dried Engineered Escherichia coli Whole-Cell Biocatalysts Enable Cell Recycling and In Vivo α-Ketoglutarate and NADPH Regeneration for Efficient Codeine Manufacture

Abstract

A recombinant whole-cell biocatalytic approach provides several advantages over isolated enzymes, such as improved enzyme stability, in vivo cofactor regeneration, and the facilitation of cascade reactions within a single cell. Whole-cell biocatalysis has gained attention in medicinal opioid production, due to its technical and economic advantages. In this study, recombinant Escherichia coli producing thebaine 6-O-demethylase (T6ODM), neopinone isomerase (NISO), and codeinone reductase (COR) were used to convert thebaine into codeine. Spray-drying was developed to optimize enzymatic activity in dried biocatalytic cells, achieving over 85% enzyme activity retention. These formulated cells wer..