Abstract: A method of separating a hydrophobic fermentation product selected from the group consisting of lipase, esterase, endoxylanase and an antibiotic from a mixture comprising said product and contaminants which method comprises adding to the mixture sequentially(1) 0.5 to 15% (w/v) of a nonionic surfactant,(2) 0.5 to 60 mg of a flocculating agent per gram of said mixture,(3) 1 to 20% (w/v) of an extra nonionic surfactant,(4) a suitable K, Na, NH.sub.4 or Mg salt selected from the group of chlorides, sulfates, acetates, carbonates or phosphates, whereby the concentration of the salt is chosen so as to have the surfactant layer on top and is between 2 and 30%;to obtain a three phase product mixture, separating the product mixture into liquid-liquid-solid fractions and recovering the hydrophobic fermentation product.