The invention discloses a preparation method of beer through mixed fermentation of zygosaccharomyces cidri and saccharomyces cerevisiae. The preparation method relates to the technical field of beer brewing, and comprises the following steps of (a) inoculating the saccharomyces cerevisiae and pichia pastoris on an inclined surface into a 12-degree P wholewheat juice, carrying out spread cultivation step by step, and collecting a yeast paste; (b) smashing malt, saccharifying, filtering, boiling, adding hops and sediments, and preparing into the 12-degree P wholewheat juice; (c) quickly coolingthe wheat juice to 10 DEG C, inoculating saccharomyces cerevisiae paste (3.0 to 6.0g/L) and pichia pastoris paste (0.01 to 1.0g/L), and carrying out main fermentation at 8 to 12 DEG C for 7 days; carrying out post-fermentation at the temperature being 0 to 4 DEG C, and storing for 5 to 7 days; filtering to obtain the beer. According to the preparation method provided by the invention, the pichia pastoris and the saccharomyces cerevisiae capable of increasing a beer fragrance flavor are utilized to be mixed and fermented for brewing the beer, so that the ester flavor substance content in the beer is increased, and the beer obtained through fermentation is excellent in quality, rich in ester flavor, harmonized in special flavor, and mellow in beer body.