Treatment of plant biomass with ozone is a promising delignification method. It was shown that lignin removal from the cell wall during ozonation was limited by topochemical reactions and toke place in the secondary rather in the primary cell wall. The separation of cellulose microfibrils, the loss of cell wall stiffness and complete removal of intercellular substance during the delignification process were visualized by SEM. The dependence of the average diameter of the cellulose microfibril aggregates in the cell wall of ozonized straw on ozone consumption was studied. Lignin removal caused an increase of size of cellulose microfibrils aggregates. It was demonstrated that there was an optimal degree of delignification, at which cellulose became more accessible to enzymes in the subsequent bioconversion processes. The data on the ozone consumption, residual lignin content, and sugars yield in the enzymatic hydrolysis of ozonized wheat straw were obtained. It was also found that the optimum delignification degree for sugars yield was ≈10% of residual lignin content and optimum ozone consumption was 2 mol·О3/mol C9PPU (phenylpropane structural unit) of lignin in raw straw.