In this study, we evaluated the association of glycerol and glucose as substrate for growth and poly(3-hydroxybutyrate) (P(3HB)) production, a biopolymer biodegradable, by the engineered strain Cupriavidus necator_glpFK against Cupriavidus necator DSM 545 (parental strain). The cultivations were carried out on a bioreactor, in batch mode. C. necator DSM 545 produced 21% of P(3HB) and the engineered strain 31%. C. necator_glpFK was able to consume all the glycerol and glucose available, presenting better performance in cell growth than the parental strain. In conclusion, the engineered strain could grow and produce P(3HB), from glycerol and glucose, more efficiently than C. necator DSM 545.