The duo was joined by vocalist and Miss Universe 2006 Zuleyka Rivera, who starred in the "Despacito" music video. Fonsi had previously told ET during rehearsals that Justin Bieber was not going to perform with them.
Nonetheless, Fonsi and Daddy Yankee pulled off a showstopper of a performance that had stars like Chrissy Teigen, John Legend and Donnie Wahlberg dancing in their seats.
The "Despacito" remix became a worldwide phenomenon, spending 16 weeks at No. 1 on the Billboard Hot 100 and tying the all-time record for consecutive weeks on top.
"Despacito" was not only a commercial but also a critical success for the collaborators. The track won four Latin GRAMMY Awards, including Record of the Year and Song of the Year, and is nominated for three GRAMMY Awards.
The song was infectious, and its music video went on to become the first YouTube video to hit 4 billion views, as well as the world's most streamed song of all time with 4.6 billion streams.
"I'm a little nervous, I'm not gonna lie," Fonsi revealed. "This is really big. Not only for me, but for Latin music in general, to be able to sing a song in Spanish for the big show. I'm really proud to be here. Being from Puerto Rico, growing up here in the United States, I've always been so proud of my roots and my language, and all these artists who are performing, they've all influenced me. So to be able to celebrate a little bit of who I am and where I'm from, and to share with the world, and break that language barrier -- especially after everything that [Puerto Rico has] been going through -- this is already just good vibes for everybody."
In December, following the GRAMMY nominations, Fonsi also told ET how proud he was of his recent accomplishments.
"I think all artists dream about achieving this at some point," he expressed. "It just makes me feel like everything I’ve been working so hard for the last two decades and all the sacrifices I have made have been worthwhile.
"I feel extremely honored and grateful to be recognized by the Academy," he added. "I’m still digesting everything that is happening, but one thing is for sure, this is historic for Latin music, and I’m proud to be a part of the selective group of artists that has crossover and shared our roots and culture with the world."
As far as achieving the same success with his new music as he did with "Despacito," he understands it was a huge feat.
"What happened with 'Despacito' is something that is hard to match, even by me, so I do not have any pressure of achieving the same," he said. "I just want to continue creating amazing music and if there is an opportunity to work with other artists from different genres, I’m all for that."
For more on how Fonsi and Daddy Yankee prepared for the big show, watch below.