Grolsch was first brewed in Groenlo in 1615 by Willem Neerfeldt. Peter Cuyper, son-in-law of Neerfeldt, was appointed master of the Grolsche brewing guild in 1676. Cuyper died in 1684, after which his descendants continued the business. Until 1876, the beer was brewed in the center of Groenlo. In that year brewery 'De Klok' built a new brewery outside the city. In 1895 it was bought by Theo de Groen, a descendant of a Zevenaar brewing family. Op 3 mei 1922 fuseerde Brouwerij De Klok uit Groenlo met de Twentse Enschedesche Stoombierbrouwerij uit Enschede. Het bedrijf ging verder onder de naam "N.V. Bierbrouwerij De Klok Enschede-Groenlo". Later veranderde de naam in Grolsche Bierbrouwerijen en werd het bier 'Grolsch' genoemd.

The De Groen family has upgraded the company from 1968 from a local brewery to an international company. Until 2004, Grolsch was brewed in Groenlo and Enschede. Since 2004, both locations have been closed and there is a new brewing factory in Boekelo-Usselo.

On 19 November 2007 the board of Royal Grolsch NV accepted a €816 million offer for the company by SABMiller. The takeover was completed with the delisting of Grolsch's shares on 20 March 2008. SABMiller subsequently sold the company to Anheuser-Busch InBev and in April 2016, the latter accepted Asahi Group Holdings Ltd.’s offer to buy not only Grolsch but also the Peroni and Meantime beer brands.