He was voiced by voice actor William Salyers.
After made a heist in Gotham Palace and tied the public along with Riddler, Joker and Catwoman, the Penugin boarded the Jokermobile to reach the chemical factory and stole the Duplicator Ray. Batman and Robin tried to stop the dastardly villains and their minions, but they escaped to a frozen food factory. Once Batman and Robin arrived, they had a fight with the villains and after the fight had finished Catwoman managed to spray them with knockout spray so the dastardly villains can make their final bat trap. The Joker tried to cook Batman and Robin with a huge oven, after Catwoman tried to make Batman a bad guy, without apparent success. But the Dynamic Duo escaped.
After leaving the factory, the Penguin and the other villains used a U.S. rocket to reach a space station built by Russia and United States. However, along Riddler and Penguin, the Joker concluded that Catwoman could betray them, because she was in love with Batman. For that reason, along with the other villains, the Joker locked Catwoman in a special room and launched her into space to kill her. However, Batman and Robin saved Catwoman from her planned death; and faced Riddler, the Penguin and the Joker in the main room of the station. After an antigravity fight, Batman defeated the villains, including Penguin, leaving them unconscious, and they were taken into custody offscreen after this.
However, in reality, the Joker, along with the Riddler and the Penguin, who had doubled themselves with the Duplicator Ray, and Batman had send to prison their duplicates. After Batman returned to his normal state, he reached to this conclusion. However, during this time, the Joker, the Riddler and the Penguin robbed important artworks from Gotham Museum, one more higher than the previous. But Batman, Robin and Catwoman appeared on the scene to stop the villains, whereby the Joker aboard the Penguin's Blimp, along the Riddler and a few henchmen from his gang. However, Batman, Robin and Catwoman defeated his henchmen along the Riddler and the Penguin. After when the Riddler was defeated shortly after, the Penguin tried to make a daring escape. He jumped off of his bimp flying down with his umbrella until Catwoman took it away with her whip making him angrily scream until he landed onto a load of fish making him more uncomfortable with his hat landed on his stomach. He was never seen in the movie after this, but it's likely that after that, he was taken into custody for his crimes with the Joker and the Riddler.
- In the trailer of Batman vs. Two-Face, he was clearly seen with the Joker and the Riddler as they have returned from Batman: Return of the Caped Crusaders. It was unknown if they will have any lines because Two-Face is the main antagonist for the sequel. Other characters like Egghead, Mr. Freeze, Shame and Clock King also appeared in the first movie as they were non-speaking characters and they return for the sequel too.