The ceremony took place at the United Nations headquarters in New York, where representatives from all 15 nations and the European Commission handed the required papers to the UN chief legal counsel.
The ratifications mean that the number of parties to the protocol is now well past the threshold of 55 needed to grant it legal status.
Now the agreement needs to be ratified by more than 55 nations who are responsible for more than 55% of greenhouse gas emissions for the law to come into force.
European commissioner for the environment, Margot Wallstrom, praised the ratification as "an historic moment for global efforts to combat climate change".
However, she warned that the pressure was now on the United States - the world's biggest polluter - to do its part.
'US should reconsider'
Conceived during the historic 1992 Earth Summit in Rio de Janeiro, the Kyoto agreement was signed in Japan in 1997.
It requires industrialised countries to reduce their greenhouse gas emissions by 8% of the 1990 levels between 2008 and 2012.
Austria, Britain, Denmark, Germany and Luxembourg are the five countries in the EU who must make the biggest cuts.
But the US repudiated the treaty, arguing that its economic interests would be threatened.
Instead of cutting emissions from 7% as required by the treaty, the Bush administration had initiated policy changes that could increase its emissions by up to 30%, the European Commission said.
"The European Union urges the United States to reconsider its position," Margot Wallstrom. "All countries have to act, but the industrialised world has to take the lead."
World Summit
United Nations Secretary-General Kofi Annan also welcomed the move, saying it was "good news for the entire world", French news agency AFP reported.
"[It is] a sound and innovative response to a truly global threat affecting rich and poor countries alike."
Since the US pullout from the treaty, the EU has been on a diplomatic offensive to ensure countries such as Russia, Japan and Canada stick with Kyoto.
Green campaigners would like to see Kyoto ratified by the World Summit on Sustainable Development later this year.