Samkelo Radebe, Zivan Smith, Arnu Fourie and Oscar Pistorius set the stadium alight as they exchanged batons – in the form of an arm tap – first in each leg of the race.
Brazil's Alan Oliveira was close to catching Pistorius on the line but Pistorius' determination won through in the end.
"Our team remained calm before the race and we each knew what we had to do," said Pistorius after the race.
"We came out here to break the world record and we smashed it.
"It makes me proud to be a part of this team with these guys tonight."
As it turned out, the Brazilians were disqualified for not touching the baton within the take-over zone as were the Americans for running out of their lane.
China's team jumped up to second place in 42.98 while Germany celebrated as they too moved into the medals with their time of 45.23.
The last event of the evening saw Teboho Mokgalagadi come second in his heat to qualify for the men's 200m T42/46 final in 27.37 seconds.
His time was sixth best overall with the adjustments made for the different classifications.
Earlier, Pistorius and Fourie stormed home in their different heats to qualify for the men's 100m T44 track final, in times of 11.18 and 11.29 respectively.
Only Britain's Jonnie Peacock ran a better time, setting a personal best of 11.08. Peacock's heat comprised US sprinter Jerome Singleton (11.46) and the Brazilian winner of the 200m final last Sunday, Alan Oliveira, who clocked 11.56.
Americans Blake Leeper, who ran second to Pistorius in 11.34, and Richard Browne, who came in behind Fourie, in 11.33, together with China's Zhiming Liu (11.84) make up the numbers for Thursday's final.
'Not my event'
Oliveira (20) pulled off a shock victory over defending champion Pistorius in the 200m on Sunday but played down his chances in the straight sprint, as had Pistorius.
"It's not really my event," Pistorius said last week. "As Jonnie [Peacock] and those guys focus on the 100, my focus is on the 400, on the complete opposite side of the spectrum when it comes to sprinting," he said.
Pistorius, the Games' biggest star after he became the first double-amputee to compete at the Olympics, remains at the centre of a row after questioning the length of his rivals' artificial running blades in his 200m defeat.
South Africa's National Paralympic Committee has written to the International Paralympic Committee (IPC) claiming that some athletes swapped their blades mid-competition, which is against the rules.
Pistorius, who has had to fight to convince the authorities that his prostheses do not give him an advantage in non-disabled races, suggested that he was at a disadvantage because his rivals were "a lot taller" in the 200m final.
Singleton on Wednesday backed calls by his rival for a review of the regulations, which currently stipulate maximum heights for prostheses through on a complex mathematical formula based on proportional body length.
The US runner, a former National Aeronautics and Space Administration (Nasa) intern, also called for a split in categories in sprint races to ensure fairness.
Pistorius and Oliveira are T43 (double below-the-knee) amputees but Singleton and Peacock are T44 (single below-the-knee) amputees.
But they race against each other because there are not enough sprinters in each category with the qualifying standards.
The IPC said this week that it is looking to introduce more single category races and field events in athletics for the next Games in Rio de Janeiro, Brazil, in 2016.
Elsewhere, Dyan Buis won the bronze medal in the men's F38 long jump, to accompany his silver in the men's 100m, and set a new world record in his class with a leap of 6.48 metres.
"I came here wanting to break the world record so I was so happy when I got it," said Buis.
"I felt it in the air because everything was in sequence and I knew it was a great jump.
Although Buis had the best jump of the night, he was competing against F37 classified athletes and the points system adjusted the distances accordingly.
"I'm happy that I'm the best in my class and I still got a medal for my country and that's something to be proud about," he said.
"It's the first time I've competed at a Paralympics and I'm first in the F38 class so that's what counts.
"Riversdale [his home town in the southern Cape] has gone crazy and I'm so glad to keep them on a high.
"I've no idea what to expect when I get home but I can't wait to see what's going to happen."
His medal takes South Africa's tally to 16 – three gold, five silver and eight bronze.
Also competing in the long jump F37/38 event was 19 year-old Andrea Dalle Ave, who finished fifth with a personal best and South African record leap of 6.02 metres in his F37 category.
"I'm very happy with my result and I think there's a lot more to come from me," said Dalle Ave.
"It is so inspiring looking around at the faces in the huge crowd and the noise which resonates when there is a British athlete competing also elevates you."
Union Sekailwe, who won a bronze medal in the 400m, failed to progress past round three of the long jump.
Zanele Situ just missed out on a medal in the women's javelin F54/55/56, finishing fourth with a best throw of 16.22 metres.
The event was won by Liwan Yang with a world record heave of 17.89 metres. – Sapa, AFP