The board was owned by the controversial King, who was such an enthusiast for chess he was engrossed in a game when a messenger told him he had been betrayed by the Scots to the Parliamentarians during the English Civil War.
The news sealed the royal's fate and he was executed on January 30, 1649.
Charles I was King of England, King of Scotland, and King of Ireland from 27 March 1625 until his execution.
It is known the King took with him two precious possessions to the scaffold where he was beheaded: a Bible and an amber games board, believed to be the one that has now sold for 601,250 pounds, the 'Daily Mail' reported.
The board, which measures 27ins by 13ins, opens into two halves, allowing it to be used for different games including chess, backgammon, draughts and Nine Men's Morris, a strategy board game which emerged from the Roman Empire.
The board would have been extremely sought after and expensive during the 17th century due to amber being found only in small quantities.
"This board was made by Georg Schreiber who was known as the 'King of the Gamesboards'.
The board is dated 1607 and was given to either James I or Henry Frederick as Charles was only seven-years-old then," said Erik Bijzet, an expert in European sculpture at auctioneers Sotherby's.
"When at the height of the Civil War a messenger arrived to inform Charles that he had been betrayed by the Scots he didn't rise from his game of chess, even though his fate had effectively been sealed," Bijzet said.
"Charles took a Bible and a games board with him to the scaffold where he was beheaded," Bijzet added.
Following his death the items were passed on to his personal chaplain, Bishop William Juxon, who read Charles his last rites.
The board then remained in his family until the 18th century before it was acquired by British peer Sir Robert Hesketh.
It has now been sold by the Second Baron Hesketh's Will Trust.
The board was bought at auction in London by a private collector, following a dramatic bidding war, for 601,250 pounds, the highest amount ever paid for an amber games board.