The big-serving South African withdrew hurt in his opening match at Melbourne Park on Tuesday.
Anderson, the only SA player in the main singles draw at the year’s first Grand Slam, had picked up a recurring knee injury in the build-up to the tournament.
Though he had withdrawn from the Chennai Open in India two weeks earlier, he turned out in the Auckland Open last week, reaching the third round.
He was trailing 2-1 after three full sets against Rajeev Ram in Melbourne, with the American holding a 3-0 advantage in the fourth, when he pulled out.
Former Australian Open finalist Raven Klaasen, the last remaining South African in the main draw, will pair up with Ram in the doubles competition later in the tournament.