Thomson Airways will perform a biofuel-powered flight on 28 July between Birmingham and Palma Mallorca using a Boeing 757 aircraft.
It will use a 50% blend of biofuel, produced from used cooking oil, and 50% kerosene in both engines. The fuel is supplied by Dutch-based company, SkyNRG.
The airline said that it will then begin weekly biofuel-powered revenue flights on the same route from September, becoming the first UK airline to do so.
It is anticipated that the weekly flights will operate for one year on the Birmingham-Palma route in the summer, switching to Birmingham-Alicante in the winter.
Thomson said that as sustainable biofuels become more commercially viable, it would expand their use across its fleet over the next three years.
Highlighting the significant premium over regular kerosene at which biofuels currently trade, Thomson Airways' managing director said: "We urge UK and EU governments to use this opportunity to review the legislation and remove the barriers around sustainable biofuels so that other airlines can follow our lead."