Just 24 hours after Rafael Nadal's first round defeat against unheralded Ukrainian Alexandr Dolgopolov, it was Wawrinka's turn to succumb to an unexpected loss at the Wimbledon warm-up event in west London.
The Swiss second seed had swept through his first round clash against Australian Nick Kyrgios in just 49 minutes as he looked to follow his memorable final triumph over Novak Djokovic in Paris with his first ever
But, in the second round, Wawrinka couldn't find a way to subdue the big-serving Anderson, who fired down 22 aces en route to his first win over a top five ranked opponent this year.
Wawrinka had been hoping to extend his winning run to nine matches, but the towering 6ft 8in Anderson had won their last three meetings, all in 2014, and he proved a thorn in the 30-year-old's side again.
A hard-fought first set looked to have swung Wawrinka's way when he secured two set points at 6-5, only for the Swiss to let Anderson off the hook as the 29-year-old raced through the subsequent tie-break.
The superstitious Wawrinka was still wearing his 'lucky' pair of garish red, white and grey checked shorts that caused a stir during his triumphant run in Paris, but his fortune took a turn for the worse in the second set.
Anderson had brought Lleyton Hewitt's Queen's farewell to a premature and painful end after saving a match point during his first round victory against the four-time champion.
He claimed a more high-profile scalp on this occasion thanks to a dramatic second set tie-break which saw Wawrinka save five match points and Anderson rescue two set points before the South African finally sealed his win.
In the quarterfinal, Anderson was to play either world number 79 Dolgopolov or Spain's Guillermo Garcia-Lopez.