目的 研究内生真菌胞外漆酶在槐米总黄酮提取中的应用。方法 内生镰刀菌在起始pH7.0的综合马铃薯培养基中,20℃下摇瓶培养6d,具有较高的漆酶活力(36.1U/mL)。槐米干粉投入真菌发酵液中进行酶解处理,比较酶料比、酶解温度、酶解时间和酶解液pH值对槐米总黄酮提取率的影响。结果 槐米干粉以酶料比40:1加入粗酶液中,在pH7.0、20℃下酶解处理1h后,总黄酮提取率可达11.4%,比常规提取法提高了28.7%。结论 内生菌漆酶辅助提取法为槐米总黄酮提取的可行新方法。

Objective To study the application of extracellular laccase from endophytic fungi to total flavonoid extraction. Methods The extracellular laccase vitality of an endophytic Fusarium sp.C-8 from Artemisa annua reached 36.1 U/mL when the fungus was cultivated in revised PDA medium with initial pH 7.0 at 20℃ on a rotary shaker incubator for 6 d. The crude laccase from the cultural medium was applied for extraction of flavonoid from the buds of Sophara japonica. The optimization on the ratio of dry buds to crude enzyme liquid,temperature,time,and pH value was carried out during the process of laccase-assistant treatment. Results After incubation with 40:1 of crude enzyme (pH 7.0) at 20℃ to dry buds for 1 h,the extraction rate of total flavonoids was 11.4 %,a more increase of 28.7 % than that in regular extraction process. Conclusion The results present a practical method of laccase-assistant extraction process on total flavones in S. japonica buds.

国家自然科学基金资助项目(30772731);江苏省自然科学基金资助项目(BK2007051);苏州大学青年教师研究基金资助项目(Q3132625)