Run by chef Daniel Humm, the Manhattan institution came out on top at the World's 50 Best Restaurants awards, widely seen as the Oscars of the food world. Spain's El Celler De Can Roca was third.
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"I don't know how I feel right now but I'm overjoyed," said an emotional Humm, who was praised for his fine dining menu inspired by the agricultural bounty of New York and its culinary traditions.
"I'm so proud to be up here. When I first started cooking 25 years ago, I never in my wildest dreams would have thought I'd find myself in this position.
"It's an unbelievable time to be a chef," he added.
The top 10 was rounded out by restaurants in France, Peru, Spain, Thailand and Austria.
The awards this year were held outside of London or New York for the first time, with Australia fast earning a reputation as a serious food destination and attracting some of the world's top chefs to open restaurants Down Under.
The move to Melbourne was part of an ongoing strategy to make the event more global, organisers said.
In other awards, Spanish eatery The Disfrutar in Barcelona was picked as the One to Watch while Ana Ros of Hisa Franko in Slovenia was named the world's best female chef.
Top pastry chef was French-born Dominique Ansel -- known as king of the cronuts -- and his bakery in New York, while a nod for sustainability went to France's Septime.
Acclaimed British chef Heston Blumenthal, whose restaurant The Fat Duck is a former winner of the top award, was handed a lifetime achievement gong.
Organisers said it was hard to think of another chef who had more of an impact on the way the world cooks.
"Such is his influence, even those that haven't been lucky enough to eat at The Fat Duck in Bray -- west of London - will likely have experienced elements of his work second-hand," they said.
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"Many of the world's greatest restaurants use methods derived from Blumenthal and his development team's ground-breaking research."
Blumenthal, who has brought his recipes to millions through TV shows that are broadcast across the world, said he was honoured by the accolade and had no plans to slow down.
"I consider the past 20 years my apprenticeship," he said. "I'm only just getting started."
The awards, run by trade magazine Restaurant, began in 2002 and have become a reference for the world's foodies, but were hit in 2015 by allegations of cosy deals between restaurants and jury members.
The jury is now made up of just over 1,000 "independent experts," including food writers, chefs, restaurant owners and gourmets.
Members list their choices in order of preference, based on where they have eaten in the past 18 months.
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