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Mr Acquaro, 54, was shot dead in the early hours of March 15, 2016 after leaving his Lygon Street cafe Gelobar and walking towards his car.

Police last year charged Vincenzo Crupi, 70, with murder. Mr Crupi on Monday appeared via video link before Melbourne Magistrates Court, which heard his lawyers wanted access to royal commission material about the police investigation into Mr Acquaro's death.

That included police theories on motive and who fired the fatal shots.

At one point magistrate Suzanne Cameron asked defence counsel Jim McGarvie if the request was being made "in relation to alternate suspects".

"Correct," Mr McGarvie replied.

Brendan Murphy, QC, appearing for Victoria Police, said police needed to assess the documents in question, but conceded there could be further material that could advantage "both the accused and the homicide squad".

The year before his death Mr Acquaro was told by police a senior mafia figure had put a contract on his life and that he was in grave danger.

Ms Cameron said on Monday it could be some time before the documents from the royal commission become available. The investigation is still at a preliminary stage.

The magistrate and lawyers did not explicitly name the royal commission on Monday, and when addressing Mr Crupi, Ms Cameron acknowledged they had "half talked in riddles and been a little bit cryptic" given the "sensitivities" over the hearings.

Mr Crupi was remanded in custody and is due to return to court on April 17.

His lawyers are likely to apply to cross-examine 19 witnesses at the substantial hearing that determines whether he faces trial.