Libya's foreign minister Moussa Koussa, one of Col Gaddafi's closest aides, has arrived in London and told officials he is quitting, the Foreign Office says.
It said Mr Koussa had indicated he was no longer willing to represent the Libyan leader's regime internationally.
The FCO added it wanted to "encourage those around Gaddafi to abandon him and embrace a better future for Libya".
A Libyan government spokesman earlier insisted Mr Koussa had been travelling abroad on a diplomatic mission.
The development came on the same day the UK took steps to expel five Libyan diplomats.
Foreign Secretary William Hague told MPs the five, who include the military attache, "could pose a threat" to UK security.
BBC political editor Nick Robinson said Mr Koussa had flown into Farnborough airport, in Hampshire, on Wednesday afternoon from Tunisia and had been debriefed, most likely by intelligence officials.
He said the fact he had flown into Farnborough was significant as he had clearly not been on a commercial flight.
"What is not clear is whether he has arrived simply to escape, or to play a wider role in any opposition to Col Gaddafi," he said.
"Clearly his defection, if that's what it becomes, will be used to urge others to follow suit and claim the regime is losing support."
Mr Koussa has not met with Prime Minister David Cameron or Foreign Secretary William Hague yet, but he is known to have been a point of contact for Mr Hague in recent weeks.
In a statement the Foreign Office said: "We can confirm that Moussa Koussa arrived at Farnborough Airport on 30 March from Tunisia.
"He travelled here under his own free will. He has told us he is resigning his post.
"Koussa is one of the most senior figures in Gaddafi's government and his role was to represent the regime internationally - something that he is no longer willing to do.
"We encourage those around Gaddafi to abandon him and embrace a better future for Libya that allows political transition and real reform that meets the aspirations of the Libyan people."
Providing armsBBC world affairs correspondent John Simpson said he suspected Col Gaddafi would shrug off Mr Koussa's resignation as he had lost other ministers in the past.
His interior minister is now in Benghazi and it did not really make any difference at all as "this is a one-horse show", he said.
Noman Benotman, a friend of Mr Koussa and a senior analyst at the UK-based Quilliam think-tank, told Reuters news agency: "He wasn't happy at all. He doesn't support the government attacks on civilians.
"He's seeking refuge in Britain and hopes he will be treated well."
Rebels opposed to Col Gaddafi's rule and fighting Libyan government forces are continuing to lose ground and are retreating from their former strongholds along the eastern coast of Libya.
Prime Minister David Cameron has said the UK is not ruling out providing arms to rebels in "certain circumstances" but no decision had been taken.
The UK has been involved in more than 160 aerial missions, as well as missile strikes, over Libya since coalition operations began on 19 March following a UN resolution.
The coalition military action is aimed at enforcing a no-fly zone over Libya and protecting civilians from attacks by forces loyal to its leader Col Muammar Gaddafi. It has denied air strikes are meant to provide cover for a rebel advance.
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