The Tankerville Arms Hotel in Fitzroy has been listed by the hotelier Mazen Tabet and his investment partners.
There are unconfirmed market expectations of around $20 million for the 1865-built pub.
He reportedly shares ownership of the Tankerville Arms with local investors Franco and Lina Rumoro.
It is on a 1,087 square metre site over three titles at 230 Nicholson Street.
It will go to auction on October 5.
Tabet paid $17 million for the Portsea Hotel in 2016 buying it from Computershare founder Chris Morris.
He also owns the Lynbrook Hotel in Melbourne's south east.
The Tankerville Arms is being marketed by Cropley Commercial's Steve Cropley and George Iliopulos in conjunction with CBRE.