Abstract: Stable isotope double labeled L-Lysine hydrochloride ¹³C₆、¹⁵N₂ was preparaed by Corynebacterium pekinense ZLD118（HS^-、AEC^r）through fermentation. The conditions of microbial fermentation and the formula of fermentation medium had been optimized. The fermentation of L-Lysine labeled with high abundance stable isotopoe ¹³C and ¹⁵N was successfully performed with the optimized process. The high abundance L-Lysine fermentation broth was purified by ion-exchange resin ，refined after decolorization by activated carbon and the final product L-Lysine hydrochloride ¹³C₆、¹⁵N₂ was got. The ¹³C abundance of the product was 98.61% and the ¹⁵N abundance was 98.15%，its purity was 98.47%. The yield of L-Lysine hydrochloride ¹³C₆、¹⁵N₂ reached 88.8%. The results showed that this process can be further used for preparation in up scale.