Brussow moves to the replacements bench, with scrum half Ricky Januarie dropping out of the matchday 22.
With Peter de Villiers and his management team opting for a five-two forwards-backs split on the bench, no reserve scrum half has been selected. Ruan Pienaar will provide scrum half cover.
The only other change to the squad that secured the first Test spoils is on the bench, where hooker Chiliboy Ralepelle replaces prop Gurthro Steenkamp.
de Villiers said: "We believe in the abilities of our players and we are confident that they will take things up a few notches on Saturday.
"We are a squad of 22 focused on a common objective, which is to go out and win a Test series against a very good Lions team.
"We know that we will be faced by a Lions team that will be playing to level the Test series and will provide us with a stern test. We must make sure that we are more than equal to the task."
The combined number of caps in the Springboks' starting line-up is 617. The squad of 22 has a combined total of 725 caps.
SOUTH AFRICA TEAM & REPLACEMENTS (v British & Irish Lions, 2009 Test series, Loftus Versfeld, Pretoria, Saturday, June 27, kick-off 3pm local time/2pm Irish time):
15. Francois Steyn (Sharks)
14. JP Pietersen (Sharks)
13. Adrian Jacobs (Sharks)
12. Jean de Villiers (Stormers)
11. Bryan Habana (Bulls)
9. Fourie du Preez (Bulls)
1. Tendai Mtawarira (Sharks)
2. Bismarck du Plessis (Sharks)
3. John Smit (Sharks) (capt)
4. Bakkies Botha (Bulls)
5. Victor Matfield (Bulls)
6. Schalk Burger (Stormers)
7. Juan Smith (Cheetahs)
8. Pierre Spies (Bulls)
16. Chiliboy Ralepelle (Bulls)
17. Deon Carstens (Sharks)
18. Andries Bekker (Stormers)
19. Danie Rossouw (Bulls)
20. Heinrich Brussow (Cheetahs)
21. Jaque Fourie (Lions)
22. Morné Steyn (Bulls)