Specialist micro-engineer Graham Short visited the Dickinson & Morris Ye Olde Pork Pie Shoppe in Melton Mowbray, Leicestershire, on Friday.
It follows the use of one of the special notes at a bakery in Kelso, Scotland, and also a cafe in Wales.
The final note will be spent this week in Northern Ireland.
Updates on this story and more from Leicestershire
Mr Short, from Birmingham, came up with the idea of engraving a 5mm portrait of Pride and Prejudice author Austen on the transparent part of the new plastic Bank of England £5 notes to mark the 200th anniversary of Austen's death next year.
He has included a different quote around each one, ensuring that each note is unique.
Anyone who finds one of the notes has been advised to contact the Tony Huggins-Haig Gallery, which launched the project.
The Bank of England's website states that it is illegal to deface bank notes under the Currency & Banknotes Act 1928.
In a statement, the gallery said it had contacted the Bank of England, which informed them that the notes were still legal tender.
Mr Huggins-Haig told BBC Scotland all of Mr Short's work has an insurance valuation of about £50,000. He also described the notes as like "the Willy Wonka golden ticket".
Mr Short's last work - a minute picture of the Queen on a pinhead - sold for £100,000.
The four notes have the following serial numbers and quotes:
Mr Short, 70, said: "As I enjoy pork pies, I thought where better to spend the English note than in Melton Mowbray?"
Stephen Hallam, from the shop, told the BBC every £5 note is not "inspected with a magnifying glass", but admitted the notes in the shop were "scrutinised" after they were told on Monday that Mr Short had visited the shop.
However, the special note could not be found.
"I'm still here making pork pies... I'm not flying off to Barbados," Mr Hallam said.
Mr Short told BBC Radio Wales' Jason Mohammad programme he chose Square Café in Blackwood High Street in Caerphilly because that was where his mother was born in 1909.
According to the Bank of England, it is against the law to "deface our banknotes (by printing, writing or impressing upon them words, letters or figures, etc.), although the question of whether or not to prosecute in individual cases is up to the police and the courts".

The suspects reportedly used automatic weapons and an anti-tank missile in the heist last September.
Their colleagues in the police helped identify them to investigators, said Dragan Lukak, interior minister of Bosnia's Serb-run region.
Two other people have been arrested for alleged involvement in the raid.
Money on board the armoured vehicle belonged to a subsidiary of UniCredit, Italy's top bank. A security guard was injured in the robbery.
Mr Lukak said the suspects, if found guilty, would have "disgraced... their unit and the whole police force".
Bosnia is administered under two entities, the Muslim-Croat Federation and the Serb Republic, each with their own police force.
One of four special £5 notes worth up to £50,000 that feature a tiny portrait of author Jane Austen has been spent in a pie shop.
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Five members of Bosnia's special police have been held over an armed raid on a bank van that netted 617,000 marka ($344,000; Â£241,000), officials say.