The Wangan and Jagalingou people are the latest to legally challenge the mine slated for Queensland's Galilee Basin.
Wangan and Jagalingou cultural leader Adrian Burragubba is appearing in the Federal Court in Brisbane on Monday to appeal a National Native Title Tribunal decision allowing the state government to issue a mining licence for the project without traditional owners' consent.
Mr Burragubba will argue Adani justified its mine in the Native Title Tribunal by exaggerating the economic benefits, including thousands of jobs it would create.
"There's been like gross overestimations of the jobs that have been offered, gross overestimations of the benefits that will come from this mine," he said.
Mr Burragubba said his people feared their homelands and cultural heritage would be destroyed if the mine went ahead and had already twice rejected a land use agreement with Adani.
They had been undermined by the native title process and Adani had not been operating in good faith, he said.
Mr Burragubba said he represented the "majority" of Wangan and Jagalingou people and their court action would be paid for by crowdfunding and helped by lawyers acting pro-bono.
"There's been like gross overestimations of the jobs that have been offered, gross overestimations of the benefits that will come from this mine"
The group is already taking action to stop the mine through the United Nations.
The court action comes after the Australian Conservation Foundation this month lodged papers against Federal Environment Minister Greg Hunt's reapproval of the mega-project.
The mine and its rail link were again conditionally approved by federal Environment Minister Greg Hunt last month.
The project hit a speed bump in August when the Federal Court set aside Mr Hunt's previous approval because the government conceded a technical error had occurred following a challenge from the Mackay Conservation Group.