title: Engineering Mycobacterium smegmatis for testosterone production.
abstract: 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. 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.
authors.affiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040, Madrid, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Fernández-Cabezón
authors.affiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040, Madrid, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Galán
authors.affiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040, Madrid, Spain.
authors.forename: José L
authors.initials: JL
authors.lastname: García
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D001203:Ascomycota / Q000201:enzymology / Q000235:genetics; D020604:Comamonas testosteroni / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D020102:Mycobacterium smegmatis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D013739:Testosterone; D015067:17-Hydroxysteroid Dehydrogenases; C032690:3-alpha-(17-beta)-hydroxysteroid dehydrogenase (NAD(+))