The final straw for the beleaguered boss was a 2-0 defeat at Hull City on Tuesday which left them third-bottom of the table, having begun the season with so much promise.
Villa's seventh loss in 10 league games, a run in which they have collected three points and scored only two goals, prompted fans to unfurl a banner calling for Lambert to be sacked.
The message was headed by the Villa board, who acted swiftly to end to Lambert's two and a half year stint at the club.
"Aston Villa Football Club has parted company this evening with manager Paul Lambert," the club said in a statement.
"First-team coach Scott Marshall and goalkeeping coach Andy Marshall will continue to prepare the squad for Sunday's FA Cup tie with Leicester City at Villa Park.
"The club would also like to place on record its thanks to Paul and take this opportunity to wish him every success in the future.
"The club will announce a new manager in due course."
A season that began with Villa taking 10 points from their first four games to sit second in the table quickly turned sour as they lost six successive games.
Since back-to back wins over Crystal Palace and Leicester City at the start of December, what little confidence they had drained away and the goals dried up.
Villa's tally of 12 goals is the lowest, by far, of any side in the top four divisions of English football. Jores Okore's effort in their 2-1 home defeat by Chelsea on Saturday ended a goal drought that had lasted almost 11 hours.
Only one team in top-flight history (Leicester with 11 in 1977-78) have scored fewer goals after 25 games than Villa this term. They have failed to find the net in 15 of their 25 league matches.
The 45-year-old Lambert, a Champions League winner as a player with Borussia Dortmund and former Scotland international midfielder, joined Villa in 2012 from Norwich City.
(Reporting by Toby Davis, editing by Ed Osmond)
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