Tony Bloom says claims that he placed bets on Brighton matches since becoming owner are "entirely false".
A story in the Guardian, external this week claimed the British billionaire was the anonymous gambler behind winnings of $70m (£52m), which allegedly included bets on Brighton.
The report linked 55-year-old Bloom, a professional gambler, to being "John Doe", who is referred to in a legal case in the US that is attempting to unmask an anonymous gambler on a lucrative winning streak.
Football Association (FA) rules prohibit club owners from placing bets on matches or competitions involving their team.
Bloom is one of a handful of owners included in a 2014 policy that allows him to continue gambling on other competitions and matches.
"I can categorically assure our supporters that I have not placed bets on any Brighton & Hove Albion matches since becoming the owner of the club in 2009," read a statement issued by Brighton on Bloom's behalf.
"I have always fully complied with these conditions, and all of my bets on football are audited by one of the world's leading accounting firms on an annual basis to ensure full compliance with the FA's policy."
Bloom, who also has a 29% share in Scottish club Hearts, said lawyers have made contact with the Guardian over the "entirely false" allegations.
"Separately, our club is in direct contact with both the Football Association and the Premier League regarding this matter," he said.
Bloom has a long history of overseeing gambling operations.
He founded online gambling website Premierbet in 2002, before selling it for about £1m in 2005.
Bloom also heads a private betting syndicate, run through his company Starlizard.

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