Using Corynebacterium glutamicum as original strain, through low energy N+ implanting into C. glutamicum, combining with isoleucine hydroxamic (Ile Hx) and other amino acid structure analogs to conduct directed screening, one high L-isoleucine yield mutant strain was obtained.After 72 h fermentation in shaking flask, the concentration of L-Ile was 21-22 g/L, and after 51 h fermentation in 50-L fermentor, the concentration of L-Ile was 32 g/L.