The £50 banknote carrying the portrait of first governor of the Bank of England Sir John Houblon will be withdrawn from circulation tomorrow.
Around 53 million of the notes with a total value of £2.6 billion are still in circulation, despite the Bank of England's announcement in January that they are to be withdrawn.
From the end of tomorrow, at midnight, only the £50 note which celebrates the 18th century business partnership of entrepreneur Matthew Boulton and engineer James Watt, who helped forge the Industrial Revolution, will hold legal tender status.
The Bank has previously advised people that if they have any Houblon £50 notes, it is best to spend, deposit or exchange them before Wednesday's cut-off point.
From next month, retailers are unlikely to accept the Houblon notes as payment, but most banks and building societies will still allow customers to deposit them into their accounts. Agreeing to exchange the notes after April 30 will be at the discretion of individual institutions.
Sir John Houblon was appointed as the Bank's first governor in 1694 and the £50 banknote celebrating him was first issued in 1994, to coincide with the Bank's 300th anniversary.
The design on the back of the note includes an image of Sir John's house in Threadneedle Street on the site of the Bank's present building.
The withdrawal of the Houblon note is part of the Bank's regular review of notes to make them more secure and crack down on fraud.
The Bank of England has issued banknotes since it was founded in 1694 and today all Bank of England notes are produced by De La Rue Currency.
The company's headquarters are in Basingstoke in Hampshire.