Emirates uses DXB as its global hub and earlier this year warned the southern runway closure would result in up to 48 Emirates aircraft not being utilised, with a 25 per cent reduction in the overall number of flights operated by the airline during the 45-day period.
More than 90 million people pass through Dubai International every year, making it the busiest airport for international traffic, ahead of London Heathrow.
Dubai Airports said the runway is nearing the end of its design life and requires a complete upgrade to accommodate future traffic growth.
Planning and preparations have been underway for almost two years, said Paul Griffiths, chief executive of Dubai Airports.
Once the runway closes tomorrow more than 90 construction vehicles enter and exit every hour, transporting more than 18,500 truckloads of concrete, asphalt, materials and personnel. More than 1900 workers will be involved in the project.
The work is planned for a quieter time of the year for travel through Dubai.