Safety evaluation and optimization of high-efficiency expression of 5hsCT in Saccharomyces cerevisiae
摘要
To evaluate the oral safety of the industrial strain Saccharomyces cerevisiae YS-5hsCT (S. cerevisiae YS-5hsCT) expressing human/salmon chimeric calcitonin (5hsCT) and optimize its fermentation conditions, providing scientific basis for industrial production of oral recombinant calcitonin. The strain demonstrated an LD50 > 10 g kg⁻1, with all toxicological tests yielding negative results and normal hematological/biochemical parameters (P > 0.05). mRNA analysis revealed optimal gene expression at 96 h, 30 °C and 225 rpm. Single-factor experiments established optimal growth conditions: 8% initial inoculum size, 72 h culture, 30 °C, 200–250 rpm agitation, and 7–8.5 l min⁻1 aeration. Under optimized conditions (10% inoculum size, 72 h, 25 °C, 225 rpm, 7 l min⁻1 aeration), the biomass reached 7.175 g l⁻1 d⁻1 and the 5hsCT protein yield was 149.24 mg l⁻1 d⁻1. These findings confirm the oral safety of YS-5hsCT and providetechnical support for industrial calcitonin production. Safety evaluations demonstrated transgenic S. cerevisiae YS-5hsCT exhibits no toxicity or mutagenicity, with optimized cultivation conditions enhancing biomass yield for potential calcitonin production applications.