The Spanish soccer club, which won the Champions League in May, is worth an estimated $3.44 billion, according to Forbes magazine.
The top three slots are occupied by football clubs, with Barcelona in second place at $3.2 billion, and Manchester United third, with a value of $2.81 billion.
Forbes calculated the value of the teams based on equity, debt and stadium deals.
MLB baseball team the New York Yankees were in fourth spot, valued at $2.5 billion.
In fifth place is NFL team the Dallas Cowboys, worth $2.3 billion.
NFL teams fill 60% of the top 50 slots, occupying 30 places.
The 20 non-NFL teams in the top 50 include six from MLB, eight from football, four from NBA basketball, one from the NHL ice hockey league and one from Formula 1 motor racing.
Forbes calculates that the average value of the top 50 teams is $1.34 billion, an 8% increase on last year.
Baseball team the New York Mets are one of three teams that dropped off this year’s list of the top 50.
The other two to fall out of the list are also familiar names, the Oakland Raiders from the NFL and F1 team McLaren.
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