George "Jug" Twitty filed divorce papers in Jefferson County Circuit Court on Dec. 29. The court filing said the couple separated Dec. 15 and have "such a complete incompatibility of temperament that the parties can no longer live together."
The Birmingham News reported the court filing Friday.
Jug Twitty declined comment on the divorce proceedings, and a phone message left for Beth Twitty on Friday was not immediately returned.
The couple attracted international attention after Holloway, then 18, disappeared in Aruba on May 30, 2005, while on a graduation trip with fellow students from Mountain Brook High School.
The Twittys appeared together during the early stages of the search. As it dragged on, Jug Twitty returned to Birmingham, and Beth Twitty remained in Aruba for almost three months.
After an unsuccessful search, Beth Twitty returned to Alabama and founded the International Safe Travels Foundation. She now spends much of her time traveling to speak to high school students.
The Twittys were married on July 26, 2000 — a second marriage for both.
Jug Twitty filed the divorce papers shortly after Beth Twitty and her daughter's father, Dave Holloway, filed suit against two former suspects in the case, Deepak and Satish Kalpoe. The suit, filed in California, claimed the Kalpoe brothers intentionally caused fatal injuries to Holloway.