%0 Journal Article
%A Guevara Acosta, Flor Govinda
%A Olortegui Flores, Yamileth
%A Fernández de las Heras, Laura
%A Perera González, Julián
%A Navarro Llorens, Juana María
%T Metabolic engineering of Rhodococcus ruber Chol-4: A cell factory for testosterone production
%D 2019
%@ 1932-6203
%U https://hdl.handle.net/20.500.14352/13790
%X Rhodococcus 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.
%~