N-(2-Deoxy-D-glucos-2-yl)-L-histidine (Glu-His), one of Heyns rearrangement products (HRPs), was prepared by condensation, dehydration and rearrangement using l-Histidine and d-Fructose as raw materials with methanol as solvent. The response surface method (RSM) was used to improve yield of product and the optimal reaction condition was as following: the original ratio of Fru:His was 1.2:1 and the temperature and time of reaction was 73.2 °C and 4.7 h, and the yield of final product was 74.10% with the purity of 99.7%. The structure of product was identified by IR, NMR and conformed as C₁₂H₁₉N₃O₇ (317.1 Da) by high-resolution mass spectrometry (HRMS) and UPLC-MS/MS. The pyrolysis behavior of Glu-His showed that its initial pyrolysis temperature was 145.2 °C and the total weight loss reached 70.61% at 800 °C. The number of pyrolysis products increased with the increase of temperature, and the main pyrolysis products were pyrans, furans, pyrazines, pyrroles, pyridines, indoles and etc. with burnt-sweet, baking, nutty, sweet and floral aroma features. At last, the fragrance enhancement effect of Glu-His in the preparation of reconstructed tobacco stem (RTS) was investigated and the result of sensory evaluation showed that the smoke of RTS cigarettes brought about more sweet and moist, less irritation, better flavor and comfort with the addition of Glu-His (0.25%, w/w).