The make-or-break vote could force the government back to the drawing board on the royalty almost two months after it took effect at five mines including Ballarat's goldmine.

The sector has been campaigning for modifications ever since it was "blindsided" in 2019 by a Labor government gold royalty announcement.

The Coalition opposition is preparing to bring the matter to a vote in the upper house, where one independent, one green and nine members of micro-parties could hold a decisive vote.

The Minerals Council of Australia is backing the bid.

"We are calling on members of the Legislative Council to stand up for Bendigo and regional Victorian jobs, so it will be interesting to see if they want to fight for a fair process," the group's James Sorahan said.

The motion only needs to pass one house of parliament because it relates to a regulation, not a bill.

That means a vote will not take place in the lower house, where it would almost certainly have been rejected by the majority Labor government.

The MCA does not oppose a royalty on Victorian gold but fears unintended consequences given that it was introduced without any consultation with industry, Mr Sorahan said.

"The impact on jobs comes down to various factors but the thing is that the royalty is tied to the gold price, not mines' profits or turnover," he said.

Instead, the royalty has been set at 2.75 per cent of market production value.

"Prices are high right now but they can't be expected to last. So if the price drops and a mine has an unsuccessful year exploring for more gold you can expect the royalty to make things hard."

The industry last year failed to convince the government to add offsets for companies exploring for new gold - a process Mr Sorahan said was the "lifeblood" of the industry and more expensive in Victoria because most mines are underground.

Even with a royalty Victoria's mines will be lucrative for miners and gold explorers, the government has argued.

It has pointed to the industry's recent renaissance, driven by the success of mines like Fosterville's which last year dug up its two millionth ounce.

Victoria is the world's only major gold producing region that does not collect a return on gold production and market prices have jumped 25 per cent since the government first announced the royalty.

Profits from the gold royalty will be invested across the state through infrastructure and services, a government spokesperson said.

"We'll keep working with the resources industry and local communities to make sure our gold industry continues to grow, but provides a fair return for Victorians," they said.