The letter, published in The Daily Telegraph newspaper, comes days after the British government seized on warnings from the Bank of England and the International Monetary Fund on the financial consequences of leaving the European Union.
"Brussels' red tape stifles every one of Britain's 5.4 million businesses, even though only a small minority actually trade with the EU," said the letter, whose signatories include Steve Dowdle, a former Sony vice-president for Europe, and David Sismey, a managing director at US investment bank Goldman Sachs.
"It is business, not government, which generates wealth for the Treasury and jobs for our communities," it added.
British Prime Minister David Cameron is fighting for his political future in the June 23 referendum, which polls suggest is neck-and-neck.
With less than six weeks until polling day, campaigning has become increasingly fractious.
Finance minister George Osborne Monday accused the Leave camp of indulging in conspiracy theories, insisting there was an "overwhelming consensus" among economists and world leaders that "Brexit", or Britain leaving the EU, would be bad for Britain.
"The next thing we know, the Leave camp will be accusing us of faking the moon landings, kidnapping (former racehorse) Shergar and covering up the existence of the Loch Ness monster," said Osborne, who was at London Stansted Airport for an announcement by Irish no-frills airline Ryanair on new investment.
"The response to the sober economic warnings from around the world by those who want to leave the EU has not been credible or serious," added Chancellor of the Exchequer Osborne.
Ryanair boss Michael O'Leary meanwhile warned that the budget airline would be forced to scale back British investment if the country votes to leave the EU, as he unveiled the creation of 450 jobs in Britain under a $1.4-billion (1.24-billion euro) boost for the firm's UK bases.
"It is exactly this type of investment that will be lost to other competitor EU members if the UK votes to leave the European Union," he said.
But the letter in The Daily Telegraph claimed that Britain's competitiveness was undermined by its membership of a "failing" 28-nation bloc.
"Outside the EU, British business will be free to grow faster, expand into new markets and create more jobs. It's time to vote leave and take back control," the letter concluded.
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