A yeast strain of Saccharomyces pastorianus no. 54 with hypoglycemic activity was isolated from soils of a winery. The aims of this study were first to investigate the effects of the cultivation conditions on proliferation and hypoglycemic activity of this yeast using the assay model of the differentiated 3T3-L1 adipocytes, and then, to confirm in vivo the hypoglycemic activity of cultured yeast by oral administration in streptozotocin (STZ)-induced diabetic mice. Among 7 diluted fruit juice samples the diluted strawberry juice (1.74 g/L reducing sugar content) was chosen as the basal medium. After investigation of the effects of addition of various substances, including 1% of 5 different sugars and glycerol, 0.1% of 6 nitrogen-containing substances, and 1 ppm of 7 growth factors, the diluted strawberry juice added with 1% glucose, 0.1% yeast extract and 1 ppm aspartic acid was optimized at 20 degrees C with initial pH value of 6.0 for cultivating S. pastorianus no. 54 in flask. The scale-up system of a 5-L fermentor was further established by using the same medium with initial pH 6.0 and being incubated at 20 degrees C with an aeration rate of 12 vvm for 96 h. The hypoglycemic activity of yeast cells cultivated in fermentor was 3.11 times of that in flask. Oral administration of the cultured yeast at a dosage of 130 mg/kg body weight/day for 6 days could significantly reduce the plasma glucose content in SE-induced diabetic mice and keep their body weights in the normal range. (C) 2011, The Society for Biotechnology, Japan. All rights reserved.