Abstract: ¹⁵N-L-leucine was prepared by fermentation on the basis of high-yielding strain TLU53-8 breeded by breeding prescription,fermentation condition and purification of ¹⁵N-L-leucine were studied. The results showed that ¹⁵N-L-leucine can be produced with more than 18 g/L. The ¹⁵N enrichment of ¹⁵N-L-leucine was more than 98%, and the product purity was more than 99%. The results demonstrated that the technique was suitable for producing high quality ¹⁵N-L-leucine.