On January 11, 2007, Couture announced his return from retirement in an interview on the Spike TV magazine show, "Inside the UFC". In a conversation with Joe Rogan, he confirmed he would face Tim Sylvia for the UFC Heavyweight Championship at UFC 68, on March 3, 2007. He also announced he had signed a four-fight, two-year deal with the UFC.
At UFC 74 on August 25, 2007, Couture successfully defended the title against Brazilian Gabriel Gonzaga, defeating him via TKO by strikes. Couture suffered a broken left arm when he blocked one of Gonzaga's kicks. The kick cleanly split his ulna, requiring him to wear a splint for six weeks.
On October 11, 2007, Couture announced he was severing all ties with the UFC, leaving two contracted fights, a position as an on-air analyst and the UFC Heavyweight Championship behind. He received $250,000 (plus $936,000 of PPV revenue) for his comeback fight with Tim Sylvia. He received a $250,000 purse for defeating Gabriel Gonzaga (plus a $35,000 "Fight of the Night" bonus and $787,000 in PPV revenue). He complained Chuck Liddell was allegedly paid more than he was, despite losing his previous two fights. Couture cited the UFC's failure to sign #1 ranked Heavyweight fighter Fedor Emelianenko, as well as disputes with UFC management, for his decision.
On October 18, 2007, UFC president Dana White said Couture remained the UFC Heavyweight Champion despite his plans to quit, and reiterated he would not release Couture from the final two fights on his contract. Couture held a press conference on October 25, 2007, where he denied his departure from UFC was a "retirement", set forth his grievances about his pay, and reiterated his belief that he would be free from any contractual obligations to the UFC after nine months. On October 30, 2007, White and Zuffa CEO Lorenzo Fertitta held another press conference, where White reiterated the UFC's position of twelve days prior, and released documents refuting Couture's claims about the pay he received.
Couture made a cameo appearance on the season finale of the CBS show The Unit, as a military guard. He played fight commentator Terry Flynn in the film Redbelt. He appeared on an episode of The History Channel show Human Weapon on September 27, 2007, and starred in the 2008 film The Scorpion King 2: Rise of a Warrior. He played Toll Road in the 2010 movie The Expendables and reprised the role in the 2012 and 2014 sequels. He did several public service announcements, one against crystal meth. He starred alongside 50 Cent and Bruce Willis in the 2011 film, Setup, as an enforcer named Petey. In 2012, he played the leading role of hard-bitten cop Paul Ross in Brandon Nutt's action film Hijacked, alongside Dominic Purcell.