Asenjo suffered the fourth such injury of his career, although the first to his left knee, during his team's 3-2 home defeat to Real Madrid on Sunday evening.
"Tests done to Sergio Asenjo have confirmed that the goalkeeper has suffered a ruptured anterior cruciate ligament to his left knee," the La Liga club said in a statement.
Asenjo, who has made 29 appearances in all competitions this season for Villarreal, will have to undergo surgery. The typical recovery time for a ruptured anterior cruciate ligament is around six months.
The 27-year-old suffered his first ACL setback in 2010 while on the books of Atletico Madrid and tore it again a year later while on loan at Malaga.
Villarreal then lost Asenjo for seven months in April 2015 after he ruptured his right ACL for the third time.
(Reporting by Adriana Garcia; Editing by Alison Williams)
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