American property tycoon Timothy Barton's bid to buy Bari collapsed on Thursday when the Serie A club's owners refused a request for a delay.
Barton's JMJ Holdings had until Wednesday to make an initial payment under the terms of a preliminary agreement but instead asked for more time to conclude due diligence.
"(We) announce that today JMJ Holdings has been told that talks over the sale of the majority stake in AS Bari have been suspended," club owners the Matarrese family said in a statement. "The Salvatore Matarrese Group will continue to offer all the possible support to Bari even in these difficult national economic times, confirming at the same time the willingness to sell the club."
The two parties held a meeting in Milan on Tuesday but the situation remained deadlocked.
Only last week, Barton said how much he was looking forward to owning Bari and said the purchase would underline a long-term commitment to doing business in southern Italy, with a renewable energy project among his plans.
Mid-table Bari, promoted back to the top flight last term, have seven points from the first six games after losing coach Antonio Conte in the close season over a disagreement about transfers.
It is the third time a bid for a Serie A club has collapsed this year.
AS Roma, a long-time takeover target, failed to come to an agreement with a Swiss consortium while Bologna were on the brink of being sold to Albanian businessman Rezart Taci before he abruptly pulled out.
Italian soccer has slipped behind the English and Spanish leagues in terms of European results in recent years with the lack of foreign investment blamed as one of the reasons.
However, many sides in Italy are owned by powerful families which have strong bonds with the clubs. AC Milan owner and Italian Prime Minister Silvio Berlusconi has repeatedly said he will not sell the club despite reports of Arab interest and their poor recent performances on the field.