The purpose of this study to determine effect of bio-emulsifier from Pseudomonas putida as feed supplement on the performance of broiler chickens. The experiment was divided in two phases. In phase 1: test of oil bubbles formation in water using bio-emulsifier, and phase 2 : in vivo test bio-emulsifier in animal. 200 one-day-old male and female broiler strain Cobb were used. On d 14, chicks were separated by sex and placed in 40 pens, sex separate (100 males or 100 females) and randomly assigned 4 treatments and 5 replicates of 5 birds each for 14 d (21d-35 d). The diets were based of corn, concentrate and rice bran meal and supplemented with: 0, 0.5,1 and 1.5 g/kg bio-emulsifier. The observed parameters were feed intake (FI), average daily gain (ADG), and feed conversion ratio (FCR). Result of experiment 1: as expected, supplementation of bio-emulsifier was increased small oil bubbles formation in medium oil in water, whereas in experiment 2 showed FI, ADG and FCR unaffected by dietary supplementation with bio-emulsifier on broiler at finisher phase (week 4 and 5) (P>0.05) and did not decrease on broiler performance. The results give new information that bio-emulsifier from Pseudomonas putida could potentially as a new alternative feed additive in the future on broiler performance.