Titled 'Etnias', the painting covers an abandoned warehouse in a street known as Olympic Boulevard in Rio de Jainero.
Brazilian artist Eduardo Kobra led the team that designed a canvas of immense hue and history, pursuing 45 days of painting to finish the mural in time for this year's Olympic Games, Guinness said.
Using a quilted design of brightly saturated figures, the mural depicts five indigenous faces of tribal people from differing continents - a choice made by the artist based on the number of rings presented in the Olympic symbol.
At 51 feet in height and stretching 560 feet in width, the illustrated faces watch over the 3 kilometre street filled with live music, bars, fireworks, and food trucks.
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"I wanted to show that everyone is united, we are all connected," Kobra said.
To create the work, the team primed the building with white paint, drew a grid-pattern to layout the design, and sprayed in brilliant colours.
"Etnias" is actually an addition to the artistic series he has begun called "Peace Outlooks" featuring spray-painted portraits of famous peaceful figures like Malala Yousafzai, Nelson Mandela and Martin Luther King Jr.
Korba has crafted street art in 20 different countries, doing renditions of Bob Dylan, Dalai Lama, Yoda and John Lennon.