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In glutamate fermentation, optimizations of production medium and fermentation parameters have been the mainly strategy for its maximization production.Response surface methodology (RSM), as an optimization strategy based on the statistical methods, was used to evaluate the effects of different organic sources of nitrogen on L-glutamate production by Corynebacterium glutamicum GDK-9.A designed RSM with different combinations of corn steep liquor and soybean protein hydrolysate was performed to attain the optimal medium of glutamate production in 5 L full-automatic fermenter.Based on the analysis result of RSM, the optimized production medium of glutamate production was obtained as follows: corn steep liquor (4.29 g/L) and soybean protein hydrolysate (10.9 g/L).Moreover, the maximum glutamate amount predicted by the second-order model was 147.17 g/L.This predicted value was in accordance with the actual experimental yield (145.73+1.24 g/L).It was concluded that the compound of corn steep liquor and soybean protein hydrolysate as the organic nitrogen sources was extremely feasible in the glutamate fermentation.