Abstract:To study the regulating effect of coenzyme on biosynthesis of antitumor alkaloid vincristine, five kinds of coenzyme were added at different concentrations. The regulating mechanism was analyzed. The results showed that adding different coenzymes independently resulted in the increase of vincristine biosynthesis. The optimal adding concentrations were confirmed. Based on the results of coenzyme one-factor experiments, five factors four levels orthogonal experiments were designed. The optimal concentration combination obtained was 1.0 mmol/L DL-thioctic acid, 1.5 mmol/L vitamin B1, 3.0 mmol/L flavin mononucleotide, 3.0 mmol/L adenosine triphosphate, and 3.0 mmol/L lysine. The highest vincristine accumulation reached 0.217 mg/g dry weight. It was the first time for successful biosynthesis of vincristine in Catharanthus roseus cell suspension culture.