A strain of high-biomass, producing glucose tolerance factor yeast strain was screened from seven yeast strains and seven strains mutated by spaceflight-induced mutagenesis through gradient concentration of chromium plates cultivation and liquid fermentation cultivation. The biomass of it was 40 g/L. Extracting GTF with ammonia and determining the content of Cr with flame atomic absorption spectrometry, the content of organic Cr was 1287 mg/g and that of total Cr was 1917 mg/g. At the same time, dynamic relationship between other parameters and the amount of organic chromium was analyzed during fermentation.