The visitors can book their spot in the playoff round with a victory, and they could also go top of the group, which would put them in a strong position to qualify for the round of 16, while the hosts have already been eliminated from the competition.
Match preview
Penarol have been typically dominant in the Uruguayan Primera Division so far this season, winning the Torneo Apertura after collecting 35 points from 15 games, six points more than bitter rivals Nacional.
However, the Uruguayan side did come unstuck in their first Torneo Clausura game last time out, falling to a 3-2 defeat away at La Luz, failing to hold on to their lead after being reduced to nine men in the second half.
Alfredo Arias's side have become accustomed to losing in the Copa Sudamericana this season, failing to pick up a single point in their four group games up to this point, which means they are now unable to progress to the last 16.
In the away trip against Sunday's opponents, Penarol succumbed to a heavy 4-1 defeat, despite having double the amount of shots and more possession throughout the match, failing to make their superiority pay.
Unlike their opponents, Defensa still have everything to play for in the Copa Sudamericana, and their progression to the playoff round will be guaranteed with a win on Tuesday, where they would take on a third-placed team from the Copa Libertadores group stage.
However, El Halcon will still be hoping they can go one step further and book their place in the round of 16 as group winners, but they will need third-placed side America MG to do them a favour against Millonarios, who they trail by one point.
Julio Vaccari's side have won three of their four group stage games up to this point, most recently defeating America MG 3-2 away from home despite trailing twice in the game, with Julian Lopez scoring the winner in the second half.
It has been a solid season for Defensa in the Argentine Primera Division so far this season, collecting 30 points from 18 games, which leaves them in a good position to quality for the Copa Sudamericana again next season.
El Halcon have recent success in the competition, being crowned champions back in 2020 after emphatically defeating Lanus 3-0 in the final.
Penarol Copa Sudamericana form:
W
L
L
L
L
Penarol form (all competitions):
W
L
W
D
L
L
Defensa y Justicia Copa Sudamericana form:
L
W
W
W
Defensa y Justicia form (all competitions):
W
D
D
L
W
D
Team News
Penarol's main attacking threat this season has been 20-year-old Matias Arezo, who has nine league goals to his name, and the youngster should lead the line once again, partnering Bruno Betancor in a 4-4-2 system.
Sebastian Rodriguez has also been key to the Uruguayan side's attacking output, scoring four goals in his last five games, and the central midfielder should get the nod alongside Sebastian Cristoforo, who found the back of the net in his last match.
In defence, Leo Coelho is likely to be handed a start, having returned to full fitness after injury, with 36-year-old Hernan Menosse also set to play.
Defensa y Justicia have no fresh injury concerns, meaning they should field a full-strength side, with Nicolas Fernandez poised to start in attack, playing just ahead of creative midfielder David Barbona in what is likely to be a 4-2-3-1 formation.
After scoring a brace in his last Copa Sudamericana outing, Gaston Togni is set to start at left-wing, with Gabriel Alanis on the opposite side and Julian Lopez in a more defensive role.
Penarol possible starting lineup:
Cardozo; Milans, Coelho, Menosse, Hernandez; Mendez, Cristoforo, Rodriguez, Rossi; Betancor, Arezo
Defensa y Justicia possible starting lineup:
Unsain; Sant'Anna, Malatini, Cardona, Soto; Escalante, Lopez, Alanis, Barbona, Togni; Fernandez
We say: Penarol 0-2 Defensa y Justicia
Considering the game means very little to Penarol, and Defensa y Justicia won the reverse fixture by a three-goal margin, we expect the visitors to emerge as comfortable victors on Tuesday evening.
Defensa have looked very solid in recent weeks, losing just one of their last 11 games in all competitions, and they have displayed they have what it takes to book their place in the next round of the Copa Sudamericana.