Aims:This study was carried out to optimize the fermentation conditions using statistical approach for polyhydroxyalkanoate(PHA) production by a local isolate, Burkholderia cepaciaBPT1213, in the shake flask system.Methodology and results:Throughout this study, B. cepaciaBPT1213 was grown in minimal salt medium (MSM) supplemented with 2% of waste glycerol (86.70% purity).The strain can produce up to 1.33 g/L cell dry weight (CDW) with 22.21% of PHA content, thus giving a total PHA concentration 0.30 g/L before optimization. A factorial design experiment that was carried out showed all parameters KH2PO4, Na2HPO4·2H2O, carbon-to-nitrogen ratio (C/N), initial pH of medium, and temperature significantly affected the growth (cell dry weight, CDW) and PHA content. Response surface methodology (RSM) using central composite design (CCD) was then applied to optimize these parameters. The optimum conditions suggested were at 2.5 g/L KH2PO4, 4.5 g/L Na2HPO4·2H2O, 30 (g/g) C/N ratio, initial medium pH of 8.5 and 37 °C cultivation temperature, with a predicted CDW of 3.43 g/L and PHA content of 45.71% contributing to 1.57 g/L total PHA concentration. The verification experiment resulted in 3.60 g/L of CDW with 48.08% of PHA content contributing to 1.73 g/L total PHA concentration.Conclusion, significance and impact of study:The statistical approach using factorial design and RSM have succeeded in increasing the production of PHA by B. cepaciaBPT1213 using waste glycerol as the sole carbon source which is a promising renewable and cheaper feedsto