Microbial synthesis of anti-tumor agent oxysophoridine through one step by filamentous fungus

Sophoridine and oxysophoridine are alkaloids extracted from Chinese traditional medicine with significant antitumor activities. Computer-aided design and preliminary structure-activity relationship studies showed that the biological activity of sophoridine can be improved after oxidation (oxysophoridine) while its toxicity significantly decreased. A new method to prepare oxysophoridine from natural product Sophoridine by biotransformation was established for the first time except for traditional extraction from plants and chemical synthesis.  Positive biocatalytic reaction was detected by TLC. Structure of conversion product was elucidated based on NMR spectroscopy. Optimizations of reactions including substrate concentration, pH, conversion time and inoculation quantity were complemented which analyzed by LC-MS. Results showed that filamentous fungus Xylariales. sp F005 (CCTCC M2014660) has the capacity of synthetizing oxysophoridine from sophoridine in the selected 37 strains. The yield of oxysophoridine reached highest at conditions pH 6.0, the third day of conversion time, 5.0 mg substrate and 5% inoculums respectively. The method for preparing oxysophoridine catalyzed by enzymes with green and sustainable synthetic processes was developed.