A joint statement put out on Twitter said the couple made the decision to part ways following "a long period of loving exploration and trial separation.
"We feel incredibly lucky to have found each other and deeply grateful for every one of the years we have been married to each other," the statement continued. "If we had known we would separate after 25 years, we would do it all again."
Bezos, who is the richest man in the world with an estimated worth of $137 billion per Bloomberg, could find himself on the precipice of a historic milestone: the costliest divorce in history.
MacKenzie Bezos is a novelist who in addition to writing is the executive director of an anti-bullying organization.
Jeff and MacKenzie Bezos launched a $2 billion fund in September to fight homelessness and fund a network of non-profit preschools in low-income communities.
The couple said they will continue to work as "partners in ventures and projects" going forward.