Die Another Day was denounced as "an insult" by a little-known group called the Solidarity of Youth and Students for Implementing the June 15 South-North Joint Declaration.
In a statement released by the North's KCNA news agency, it said the 20th Bond movie "represents the real intention of the US - keen on war as it considers the North as part of an axis of evil".
Last month, North Korea called on the US to stop showing the movie in which Bond - played by Pierce Brosnan - is tortured by North Korean agents.
In one scene he tries to stop an illegal arms deal between a South African diamond smuggler and an evil North Korean officer.
The North Korean government claimed the film was "dirty and cursed burlesque".
It has also upset South Koreans, especially a scene in which Bond has sex in a Buddhist temple.
In the latest condemnation, North Korea claimed the film caused "division and confrontation between the South and the North" and "makes mockery of the Korean nation".
"If the US continues showing the movie despising the Korean nation, all the people will turn out in a fiercer anti-US struggle," it said.
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"The US... spreads abnormality, degeneration, violence and fin-de-siecle corrupt sex culture"
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North Korean government spokesman
The group releasing the statement takes its name from the date of a 2000 summit between North and South Korean leaders in which they vowed to work towards reunification and end half a century of hostility.
During a similar condemnation in December, North Korea said the film proved the US was "the root cause of all disasters and misfortune of the Korean nation" and was "an empire of evil".
The US was "the headquarters that spreads abnormality, degeneration, violence and fin-de-siecle corrupt sex culture", it added.
In the movie, which co-stars Halle Berry, Korea is shown with a farmer tilling a field with a cow - which critics say makes the country appear backward.