The Wal-Mart on Grey Lag Way in Lexington is among stores nationwide cutting its overnight hours.
A sign on the door of the Hamburg location states that beginning July 26, the store will close at midnight and reopen at 6 a.m.
No other Lexington Wal-Marts are changing their hours.
The move is part of the Arkansas-based retailer's closing of some of its 24-hour supercenters for at least a few hours each night. The time will be used to stock shelves and organize stores, officials said.
About 40 stores are affected, Wal-Mart spokesman Brian Nick said. Other stores that will cease 24-hour status are in Philadelphia, New Jersey and Maryland.
About two dozen 24-hour stores previously had their hours reduced in the spring.