The WBO said it is against rules for boxers to hold world titles in multiple weight classes and that Mayweather had to indicate which weight division he would retain.
Mayweather, who finished ahead on all three scorecards of what proved to be the top-grossing prize fight of all-time, had until 4:30 p.m. ET (2030 GMT) last Friday to advise the WBO of his position.
"The WBO World Championship Committee is allowed no other alternative but to cease to recognise Mr. Floyd Mayweather, Jr. as the WBO Welterweight Champion of the World and vacate his title, for failing to comply with our WBO Regulations of World Championship Contests," the WBO said in a statement.
Mayweather, who earned over $200 million from the fight which generated a record pay-per-view revenue in the United States of $400 million, has two weeks to launch an appeal.
(Reporting by Frank Pingue in Toronto; Editing by Ian Ransom)
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