关键词: 芋螺毒素, 大肠杆菌, 发酵条件, 优化

Abstract: Recombinant conotoxins could be simply synthesized by genetic engineering, and fermentation conditions affect its expression yield. To obtain high expression, the fermentation conditions for engineered E. coli were optimized by using single factor test methods, such as fermentation time, IPTG concentration, initial pH and fermentation temperature. Expression of total protein in the supernatant was measured using a Protein Assay Kit, which was separated in Tricine-SDS-PAGE and the amount of recombinant conotoxins was analyzed by scanning grayscale analysis. The results showed that the optimum fermentation conditions were as follows: fermentation time was 4 h, IPTG concentration was 0.4 mmol/L, initial pH was 6.0, and the fermentation temperature was 26℃, the expression yield was 18 mg/L after optimization. The results laied a solid foundation for large-scale production of conotoxin MrVIB.

Key words: conotoxin, Escherichia coli, fermentation condition, optimization