The Leighton Buzzard Superstore is one of two stores to cease 24-hour shopping in Bedfordshire with the Flitwick Superstore also reverting back to closing at midnight.
Nearby Buckingham is another which will stop 24-hour and the trio form part of a wider scheme affecting 76 stores across the UK.
The company says the growth of online shopping means that certain shops had few customers during the night.
Normal night time activities - such as refilling shelves - will continue in the supermarkets so Tesco says the impact on employees will be “minimal”.
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The move is Tesco’s latest effort to turn around its business which has been hit by challenges on several fronts.
The outlets affected will close between midnight and 6am and 400 staff will be “affected”.
Tesco says it will do its best to make sure all staff affected are “found a new position either in their existing store or stores local to them”.
There are currently about 400 Tesco stores open 24 hours a day.
“It makes much more sense for colleagues at those stores to focus on replenishing the shelves instead and making sure they’re fully stocked when they open their doors at 6am,” said Tesco retail director Tony Hoggett.
The move is the latest step in Tesco supremo Dave Lewis’ shake-up of the supermarket’s business.
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