Tequila agave bagasse (TAB) is the fibrous waste from the Tequila production process. It is generated in large amounts and its disposal is an environmental problem. Its use as a source of fermentable sugars for biotechnological processes is of interest; thus, it was investigated for the production of polyhydroxybutyrate (PHB) by the xylose-assimilating bacteria Burkholderia sacchari. First, it was chemically hydrolyzed, yielding 20.6 g·L^-1 of reducing sugars, with xylose and glucose as the main components (7:3 ratio). Next, the effect of hydrolysis by-products on B. sacchari growth was evaluated. Phenolic compounds showed the highest toxicity (> 60% of growth inhibition). Then, detoxification methods (resins, activated charcoal, laccases) were tested to remove the growth inhibitory compounds from the TAB hydrolysate (TABH). The highest removal percentage (92%) was achieved using activated charcoal (50 g·L^-1, pH 2, 4 h). Finally, detoxified TABH was used as the carbon source for the production of PHB in a two-step batch culture, reaching a biomass production of 11.3 g·L^-1 and a PHB accumulation of 24 g PHB g^-1 dry cell (after 122 h of culture). The polymer structure resulted in a homopolymer of 3-hydroxybutyric acid. It is concluded that the TAB could be hydrolyzed and valorized as a carbon source for producing PHB.