RESULTS: In this study, we use the crude enzymes from engineered Saccharomyces cerevisiae to directionally convert TCDCA from chicken bile powder to TUDCA at the committed ratio in vitro. This S. cerevisiae strain was modified with heterologous 7α-hydroxysteroid dehydrogenase (7α-HSDH) and 7β-hydroxysteroid dehydrogenase (7β-HSDH) genes. S. cerevisiae host and HSDH gene combinatorial optimization and response surface methodology was applied to get the best engineered strain and the optimal biotransformation condition, respectively, under which 10.99 ± 0.16 g/L of powder products containing 36.73 ± 6.68% of TUDCA and 28.22 ± 6.05% of TCDCA were obtained using 12.00 g/L of chicken bile powder as substrate.
CONCLUSIONS: This study provides a healthy and environmentally friendly way to produce potential alternative resource for bear bile powder from cheap and readily available chicken bile powder, and also gives a reference for the green manufacturing of other rare and endangered animal-derived valuable resource.
结果:在这项研究中,我们使用来自工程化酿酒酵母的粗酶在体外以确定的比例将TCDCA从鸡肉胆汁粉末定向转化为TUDCA。用异源7α-羟基类固醇脱氢酶(7α-HSDH)和7β-羟基类固醇脱氢酶(7β-HSDH)基因修饰该酿酒酵母菌株。采用酿酒酵母宿主和HSDH基因组合优化和响应面法,得到最佳工程菌株和最佳生物转化条件,分别,以12.00g/L鸡胆汁粉为底物,得到10.99±0.16g/L含36.73±6.68%TUDCA和28.22±6.05%TCDCA的粉末产品。
结论:这项研究提供了一种健康环保的方法,可以从廉价易得的鸡肉胆汁粉中生产熊胆粉的潜在替代资源,也为其他珍稀濒危动物宝贵资源的绿色制造提供了参考。