Biden cited a strategic need to protect domestic industry - but the move drew sharp criticism from both companies.
The decision came after a government panel failed to reach consensus last month on whether the acquisition threatened US national security, shifting the decision to the veteran Democrat in the waning days of his presidency.
The move nevertheless enjoyed rare bipartisan agreement, with Republican President-elect Donald Trump and incoming vice president JD Vance also campaigning against the sale.
"This acquisition would place one of America's largest steel producers under foreign control and create risk for our national security and our critical supply chains," Biden said in a statement.
"That is why I am taking action to block this deal."
The United Steelworkers union quickly welcomed the announcement.
"We're grateful for President Biden's willingness to take bold action to maintain a strong domestic steel industry and for his lifelong commitment to American workers," USW International President David McCall said.
Nippon Steel and US Steel expressed disappointment with the outcome, saying it "reflects a clear violation of due process and the law."
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