Guevara Acosta, Flor GovindaOlortegui Flores, YamilethFernández de las Heras, LauraPerera González, JuliánNavarro Llorens, Juana María2023-06-172023-06-172019-07-26Guevara G, Olortegui Flores Y, Fernández de Las Heras L, Perera J, Navarro Llorens JM. Metabolic engineering of Rhodococcus ruber Chol-4: A cell factory for testosterone production. PLoS One. 2019 Jul 26;14(7):e0220492. doi: 10.1371/journal.pone.0220492. PMID: 31348804; PMCID: PMC6660089.1932-6203https:// doi.org/10.1371/journal.pone.0220492https://hdl.handle.net/20.500.14352/13790Rhodococcus ruber Chol-4 is a potent steroid degrader that has a great potential as a biotechnological tool. As proof of concept, this work presents testosterone production from 4- androstene-3,17-dione by tailoring innate catabolic enzymes of the steroid catabolism inside the strain. A R. ruber quadruple mutant was constructed in order to avoid the breakage of the steroid nucleus. At the same time, an inducible expression vector for this strain was developed. The 17-ketoreductase gene from the fungus Cochliobolus lunatus was cloned and overexpressed in this vector. The engineered strain was able to produce testosterone from 4-androstene-3,17-dione using glucose for cofactor regeneration with a molar conversion of 61%. It is important to note that 91% of the testosterone was secreted outside the cell after 3 days of cell biotransformation. The results support the idea that Rhodococcus ruber Chol-4 can be metabolically engineered and can be used for the production of steroid intermediates.engAttribution-NonCommercial-NoDerivatives 4.0 Internationaljournal articlehttps:// doi.org/10.1371/journal.pone.0220492https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0220492open access577.175.62579.222.6Rhodococcus Chol-4TestosteroneMetabolic engineeringSteroidBioquímica (Biología)Microbiología (Biología)2302 Bioquímica2414 Microbiología