The invention provides a preparation method of reconstituted tobacco. The preparation method comprises the following steps of: 1, uniformly mixing tobacco stems, tobacco
powder and water, then adding a
cell disruption
enzyme and an assistant I for carrying out
enzymatic hydrolysis; 2, heating and extracting a mixture in the step 1, carrying out
enzyme deactivation, then filtering to obtain filtrate and filter residue, concentrating the filtrate to obtain a concentrated solution; 3, adding a
mixed culture and an assistant II in the concentrated solution, carrying out
fermentation and
enzymatic hydrolysis to obtain a tobacco extraction solution; 4, adding
slurry in the filter residue in the step 1, uniformly mixing, adding
hydrogen peroxide for carrying out oxygenolysis, then adding
catalase for hydrolyzing residual
peroxide to obtain mixed
slurry; and 5, preparing the mixed
slurry into a slice paper base,
coating the tobacco extraction solution on the slice paper base,
drying and preparing into the reconstituted tobacco. According to the preparation method of the reconstituted tobacco, the
mixed culture is fermented in the concentrated solution, and thus the content of
lignin can be reduced, and the smoking effect is improved.