A replacement had to be found for Manchester City's Jesus, who has returned home after suffering a fractured eye socket in Brazil's 1-0 Superclasico loss to Argentina at the MCG on Friday.
It will be Souza's chance to impress team manager Tite ahead of next year's World Cup in Russia and represents his first opportunity to start for Brazil after late substitute appearances in qualifiers against Uruguay and Paraguay this year.
"For me it's great to be called up and to have the opportunity to play," said Souza, who normally plays as an attacking midfielder for Sport Recife.
"It will be important to show my best. There are so many centre-forward options in Brazil: Fred, Firmino, Gabriel Jesus, are all looking for a place in the team.
"I've have the dream of maybe being at a World Cup, it's difficult, they're very good players, the competition is great, but I'm enjoying it."
Along with Souza, Brazil made significant team changes with Tite announcing that Chelsea defender David Luiz will start as the holding midfielder, with Paulinho and Coutinho in front of him.
Thiago Silva will be partnered in defence by Rodrigo Caio, with Alex Sandro (Juventus) and Rafinha (Bayern Munich) at fullback and Diego Alves in goal.
Tite’s front three will include Douglas Costa, Giuliano and Souza.
Liverpool midfielder Coutinho will captain his country for the first time against the Socceroos.
Tite continued the rotation of captains in the national team with Coutinho to take over the captain's armband after Thiago Silva led the team against Argentina on Friday.
"I have to give them the chance to play," Tite said at his pre-game press conference Monday.
"This is it – the opportunity for everyone to play. My sense of justice is to give everyone an opportunity.
"I can think just about me, winning the matches and having a good (winning) record but then someone doesn’t have an opportunity.
"I have to give them opportunities and even if we lose matches, they know I'm watching them. I will make some mistakes but I’m trying to do it right.
"This is the time to give them their opportunities."
Brazil, who have already qualified for Russia 2018, did not bring leading stars Neymar, Marcelo, Dani Alves and Roberto Firmino to Australia.
The Socceroos are coming off a 3-2 home win over Saudi Arabia last Thursday which kept alive their automatic qualifying hopes for next year's World Cup finals.
Australia are using the prestige Brazil friendly as their final match preparation for this month's Confederations Cup in Russia.
"Anytime that kind of opportunity comes along you want to make the most of it," Australia coach Ange Postecoglou said.
"I guess the key is not to get too caught up in who the opponent is but make sure you get something out of the exercise."
Former Everton star Tim Cahill is expected to get some game time against Brazil.