内生链格孢菌纤维素酶辅助提取丹参茎叶中总丹酚酸条件优化  

Optimization of extraction conditions of total salvianolic acid from stems and leaves of Salvia miltiorrhiza by cellulase from endophytic Alternaria

作　　者：张芮 刘水永恒 孙士妍 张可轩 汪玉 田园 史仁玖 常正尧 李艳玲 ZHANG Rui;LIU Shui;Yong-heng;SUN Shi-yan;ZHANG Ke-xuan;WANG Yu;TIAN Yuan;SHI Ren-jiu;ChANG;Zheng-yao;LI Yan-ling(College of Life Sciences,Shandong First Medical University&Shandong Academy of Medical Sciences,Taian 271016,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)生命科学学院,山东泰安271016

出　　处：《泰山医学院学报》2020年第3期172-177,共6页Journal of Taishan Medical College

基　　金：国家自然科学基金(81403036);山东省自然科学基金(ZR2014HM048);山东省重点研发计划(2016GSF202002);山东省中医药科技发展计划(2017-239);国家大学生创新创业训练计划(201610439259,201710439317)。

摘　　要：目的研究内生真菌胞外纤维素酶在丹参茎叶总丹酚酸提取中的应用。方法内生链格孢菌在液体产酶培养基中,28℃下摇瓶培养7 d,具有较高的纤维素酶活力(46.63 U/mL)。丹参茎叶干粉投入真菌发酵液中进行酶解处理,利用Design-Expert 8.0.6软件釆用三因素三水平响应面分析法,优选丹参内生真菌纤维素酶提取总丹酚酸最佳提取工艺条件。结果利用内生链格孢菌纤维素酶提取丹参茎叶总丹酚酸的最佳提取工艺条件为:加酶液量80 mL,酶解pH 7.0,提取时间6 h,模型相关系数R2=0.9257,拟合度较好。在该条件下,丹参茎叶中总丹酚酸的提取率3.72 mg/g,与理论提取率(3.69 mg/g)接近。结论内生菌纤维素酶辅助提取法可以充分提取出丹参茎叶中的总丹酚酸,为丹参的工业化生产及中药废弃物的再利用提供了新方法。Objective:To apply extracellular cellulase of endophytic fungi in extraction of total salvianolic acid from stem and leaf of salvia miltiorrhiza was studied.Methods:The extracellular cellulase vitality of an endophytic Alternaria sp.from Salvia miltiorrhiza reached 46.63 U/mL when the fungus was cultivated in liquid enzyme production medium at 28℃on a rotary shaker incubator for 7 d.The stem and leaf dry powder of salvia miltiorrhiza was put into the fungal fermentation liquid for enzymatic hydrolysis treatment,and the three-factor and three-level response surface analysis method was adopted by design-expert 8.0.6 software to optimize the optimal extraction conditions for total salvianolic acid extraction by cellulase from endophytic fungi of Salvia miltiorrhiza.Results:The optimal extraction conditions of total salvianolic acid from stems and leaves of salvia miltiorrhiza by cellulase of endophytospora were:80 mL of enzyme solution was added,pH of enzymatic hydrolysis was 7.0,extraction time was 6 h,model correlation coefficient R2=0.9257,and the fitting degree was good.Under these conditions,the extraction rate of total salvianolic acid from stem and leaves of salvia miltiorrhiza was 3.72 mg/g,which was close to the theoretical extraction rate of 3.60 mg/g.Conclusion:The total salvianolic acid can be fully extracted by this cellulase assisted extraction method,which provides a new method for the industrial production of Salvia miltiorrhiza and the reuse of traditional Chinese medicine waste.

关 键 词：丹参 内生真菌 纤维素酶 丹酚酸 响应面法 

分 类 号：R915[医药卫生—微生物与生化药学]