The move sets up a court battle between Brussels and the internet giant that could take years to resolve and make already tense relations between Europe and the US tech giant even more fraught.
The EU hit Google with the record fine in June for illegally favouring its shopping service in search results, giving the company 90 days to comply or face further fines.
A spokesman for the Court of Justice of the European Union confirmed to AFP that the appeal had been submitted.
Concurrent to the appeal, Google last week also notified to the EU that it would attempt to meet the demands of the EU decision.
In an interview with AFP, Europe's anti-trust chief Margrethe Vestager last week said some of that proposal "pointed in the very right direction", but could only be judged once in effect.
This had raised hopes that Google may be choosing a more conciliatory road with the EU, but the appeal risks embittering matters still further.
The legal battle extends the already seven-year tussle with the EU, including a failed effort to settle, that could affect crucial parts of Google's business model.
The fine over Google Shopping broke the previous European Union record for a monopoly case against US chipmaker Intel of 1.06 billion euros in 2009 and made the EU the global leader in regulating Silicon Valley giants.
Brussels accuses Google of giving its own service too much priority in search results to the detriment of other price comparison services, such as TripAdvisor and Expedia.
The EU is also expected to soon decide another case against Google over abusing its dominance of internet search to impose its Android mobile operating system.
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