The tech executive and social media leader achieved the feat with total net worth that Forbes estimated at $44.6 billion, up from $33.4 billion in the 2015 ranking.
Zuckerberg is among 1,810 billionaires who made the updated list, with aggregate net worth of $6.48 trillion, down from $7.05 trillion last year, Forbes said.
Microsoft founder Bill Gates maintained the top spot as the world's richest person. His estimated $75 billion net worth, down from $79.2 billion last year, gave him the number one position for the 17th time in the last 22 years, Forbes said.
Spanish clothing retailer Amancio Ortega, best known in the U.S. for the Zara fashion chain, moved up two spots to second place as his estimated net worth increased $2.5 billion to $67 billion, the listing showed.
Famed business investor Warren Buffett kept third place in the ranking even though his net worth dropped $11.9 billion to an estimated total of $60.8 billion.
Mexican telecom magnate and investor Carlos Slim Helú ranked fourth, down two spots from last year, as his estimated net worth fell to $50 billion from the $77.1 billion Forbes reported in 2015.
Amazon CEO and founder Jeff Bezos moved up to fifth place from the 15th spot last year as his net worth increased to $45.2 billion.
Rounding out the new top ten are Larry Ellison, executive chairman of software giant Oracle ($43.6 billion); Michael Bloomberg, a former New York City mayor who heads the business information firm that bears his name ($40 billion); and brothers Charles and David Koch ($39.6 billion), who lead U.S. multinational corporation Koch Industries.
The new ranking is also notable for those who didn't make this year's list.
J. Michael Pearson, CEO of Valeant Pharmaceuticals International, fell out of the rankings as the Canada-based drugmaker lost roughly 75% of its value over the last six months amid government and regulatory investigations of the company's pricing and marketing practices.
U.S. fashion designer Tory Burch also missed the 2016 list, as the value of luxury fashion brands fell, Forbes said.
In all, 190 women made the 2016 billionaire ranking, seven fewer than last year. Leading the women is France's Liliane Bettencourt, the businesswoman, heiress and grand dame of cosmetics giant L'Oréal. She took the 11th spot overall with an estimated net worth of $36.1 billion, Forbes said.
The net worth of Christy Walton, widow of one of Walmart founder Sam Walton's sons, fell from an estimated $36.5 billion last year to $5.2 billion in 2016 based on what Forbes said was new details of how her fortune is split with her son, Lukas. That dropped Walton from the top-ranked woman billionaire last year to the 258th spot in 2016, the magazine said.
The new billionaire list also showed that:
India's richest are getting poorer, according to Forbes, as falling stock prices, corruption scandals in Asia's third-largest economy and a global slowdown wiped 20 percent off the total value of the country's 100 wealthiest in the last year.
10 years, 1 month ago