A bio-fluorescence method for the determination of steroid hormone in environment substance was established. Enzyme can specifically recognize steroid ring structure of hormone-steroid. Green fluorescent protein (GFP) has a report-phenomeon. On the base of these two features, a bio-induced fluorescence detection system was established through the genetic homologous recombination techniques, the research achieved the specificity and sensitivity of the monitoring of steroids. After preparing GFP mutant strains and cell-free system working solution containing 1. 0 g/L protein, the estradiol standard steroid in different concentrations was detected at room temperature. The results show that the fg level of steroid hormone can be detected within 10 min at room temperature. The detection time can maintain stable in 30 min. The linear range is 0. 001 - 0. 266 fg/mL. This method is more sensitive than HPLC with six orders of magnitude. The method is sensitive, simple, accurate, specificity, and can be applied to the inspection of steroid in environment substance.
