Abstract The octapeptide Lys-Gly-Asp-Glu-Ser-Leu-Ala, also called beefy meaty peptide (BMP), has previously been demonstrated to be a savory seasoning. In order to produce BMP in pilot scale, recombinant P. pastoris GS115-16B2 secreting the fusion peptide containing 16 tandem repeated BMP was inoculated in a 500L fermentor to carry out a three-step fermentation process, i.e., glycerol batch phase and a glycerol fed-batch phase to achieve high cell densities, followed by a methanol fed-batch induction phase. The cells were grown to desired cell density on glycerol at 30°C and pH 5.0 and then induced on methanol for expression of BMP at 28°C and pH 3.0. Dissolved oxygen (DO) was maintained above 20%-35% saturation by adjusting agitation rate, aeration and fermentor pressure. Samples were analyzed to determine the cell density (OD600), wet cell weight and BMP production. It is found that，after the culture for 118 hours, the cell density(OD600), wet cell weight and the yield of target protein are respectively 184.5, 318.7g/L and 172 mg/L.
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