Engineering Mycobacterium smegmatis for testosterone production.

Lorena Fernández‐Cabezón,José L. García,Beatriz Galán

doi:10.1111/1751-7915.12433

Abstract

SummaryA new biotechnological process for the production of testosterone (TS) has been developed to turn the model strain Mycobacterium smegmatis suitable for TS production to compete with the current chemical synthesis procedures. We have cloned and overexpressed two genes encoding microbial 17β‐hydroxysteroid: NADP 17‐oxidoreductase, from the bacterium Comamonas testosteroni and from the fungus Cochliobolus lunatus. The host strains were M. smegmatis wild type and a genetic engineered androst‐4‐ene‐3,17‐dione (AD) producing mutant. The performances of the four recombinant bacterial strains have been tested both in growing and resting‐cell conditions using natural sterols and AD as substrates respectively. These strains were able to produce TS from sterols or AD with high yields. This work represents a proof of concept of the possibilities that offers this model bacterium for the production of pharmaceutical steroids using metabolic engineering approaches.

Highlights

Testosterone (TS) is one of the oldest drugs used in medicine and has a long efficacy and safety record for hormone replacement therapy in men with androgen deficiency
A new biotechnological process for the production of testosterone (TS) has been developed to turn the model strain Mycobacterium smegmatis suitable for TS production to compete with the current chemical synthesis procedures
This work represents a proof of concept of the possibilities that offers this model bacterium for the production of pharmaceutical steroids using metabolic engineering approaches

Summary

Introduction

Testosterone (TS) is one of the oldest drugs used in medicine and has a long efficacy and safety record for hormone replacement therapy in men with androgen deficiency. TS has not been detected as a metabolic intermediate when M. smegmatis mc2155 is cultured in the presence of phytosterols or cholesterol, neither in the wild-type strain nor in the AD-producing strain (Galan et al, unpublished), unlike in other mycobacterial species (Wang et al, 1982; Smith et al, 1993; Egorova et al, 2002b). This observation suggests that M. smegmatis mc2155 does not contain a functional gene encoding a 17b-HSD or at least, it is not induced in the presence of these compounds

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