The divorce settlement could prove to be the biggest of all time.
Bezos is worthmore than $137 billion, according to Bloomberg, making him the richest person in the world.
According to Forbes' real-time ranking of the world's richest billionaires, Bezos is worth $136.6 billion ($A190bn). The business man also owns the Washington Post. An Amazon spokesman confirmed the divorce.
MacKenzie is a novelist and together they have four children.
"The property acquired during the marriage is common property", said Jennifer Payseno, a family lawyer at the firm McKinley Irvin in Seattle.
Bezos founded Amazon in his garage in 1994 and has remained through the company since its inception.
Amazon didn't immediately respond to a request for comment.
It turned ugly and stretched on for years after Elaine was granted nearly half of Steve's ownership in the company back when they divorced in 2010.
"For the most part, they're going to want to keep things private and quiet", she said. While Oracle Corp.'s Larry Ellison has been through multiple divorces, none has affected his stake in the software maker.
Amazon's stock edged up $US2.84 Wednesday to close at $US1659.42. MacKenzie was one of the company's first employees.
In spite of their separation and impending divorce, Bezos wrote that he and MacKenzie will continue to share their lives in many ways.
The couple married in 1993 after they met at hedge fund DE Shaw in NY, where they both worked. The two were married in 1993.
Jeff Bezos, CEO of Amazon and McKenzie Bezos attend the Amazon.com Sundance Party in Park City, Utah, Jan. 22, 2005.
If Jeff Bezos signed a prenup, his fortune is safe.
"My office was next door to his, and all day long I listened to that fabulous laugh", MacKenzie Bezos told Vogue in a 2013 profile, explaining how they met. She also runs a company called Black Ops Aviation, which specializes in shooting aerial shots for TV shows, movies and commercials.