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The Medway Tunnel is set to shut less than a month after a previous closure sparked traffic "chaos".
From 8pm tomorrow the busy route, which links Chatham and Strood, will be shut for engineering work.
The closure will last until 5.45am on Wednesday.
Last month a systems failure forced the tunnel to close for emergency repairs leaving drivers to face gridlock for two days.
The issue resulted in a loss of critical functions that maintain safety within the tunnel.
Affected by the incident were the systems that detect incidents, CCTV coverage, and other functions such as ventilation fans, lighting and pumps.
A spokesman for Medway council said: "The planned closure will enable engineers to further work on the cable which was identified as causing the technical systems failure in December.
"The closure has been scheduled for overnight to minimise disruption for motorists."