NOVELTY - Method for producing high yield of 2-keto-L-gulonic acid (2-KLG), involves (a) selecting strains chosen from Ketogulonigenium vulgare and Bacillus megaterium, (b) preparing culture medium chosen from solid culture medium, liquid seed culture medium and initial fermentation culture medium, (c) culturing strains in flask at 29 degrees C for 24 hours under shaking condition at a speed of 190 revolutions/minute (rpm), and (d) supplementing materials to the fermentation tank in stages and controlling the supplementation of materials during fermentation.    USE - The method is useful for producing high yield of 2-KLG.    ADVANTAGE - The method is simple to perform, cost-effective and environmentally-friendly, regulates physiological conditions of two strains in fermentation of 2-KLG, provides environmental and nutritional conditions that are more suitable for growth of two strains, increases biosynthesis rate of the two strains effectively, and saves energy.    DETAILED DESCRIPTION - Method for producing high yield of 2-keto-L-gulonic acid (2-KLG), involves (a) selecting strains chosen from Ketogulonigenium vulgare and Bacillus megaterium, (b) preparing culture medium chosen from (i) solid culture medium (pH 7) comprising (in wt.%) sorbose (1.5), yeast extract (0.2), corn residues (0.5), urea (0.2), magnesium sulfate heptahydrate (0.03), potassium dihydrogen phosphate (0.02), calcium carbonate (0.5) and agar (2.5), (ii) liquid seed culture medium (pH 7) comprising sorbose (1.5), yeast extract (0.3), corn residues (0.5), urea (0.2), magnesium sulfate heptahydrate (0.01), potassium dihydrogen phosphate (0.02), and calcium carbonate (0.5), and (iii) initial fermentation culture medium (pH 6.7-7) comprising sorbose (2 at initial stage and 25 during the process), corn residues (0.8-1.1), urea (0.1-0.2), magnesium sulfate heptahydrate (0.02) and potassium dihydrogen phosphate (0.05), (c) culturing strains in a flask at 29 degrees C for 24 hours under shaking condition at a speed of 190 revolutions/minute (rpm), and (d) supplementing materials to the fermentation tank in stages and controlling the supplementation of materials during fermentation by controlling the temperature at 28-33 degrees C for 0-32 hours during strain culture, controlling the temperature at 34-38 degrees C during strain culture from 32nd hour to the end of fermentation, supplementing 0.2-0.5% of corn residues during strain culture for 16-20 hours, and supplementing 0.1-0.35% of urea during strain culture for 28-32 hours.