酶法转化柚皮苷产普鲁宁工艺研究  

Enzymatic Preparation of Prunin from Naringin with Pre-treated Naringinase

作　　者：杨秋明[1,2,3,4] 王之路 倪辉[1,2,4] 蔡慧农[1,2,4] 李利君[1,2,4] 肖安风[1,2,3,4] 

机构地区：[1]集美大学食品与生物工程学院,福建厦门361021 [2]福建省食品微生物与酶工程重点实验室,福建厦门361021 [3]福建省海洋功能食品工程技术研究中心,福建厦门361021 [4]厦门市食品与生物工程技术研究中心,福建厦门361021

出　　处：《中国食品学报》2016年第5期68-75,共8页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31271914);福建省科技计划重点项目(2011N1008);厦门科技计划项目(3502Z20153008)

摘　　要：优化柚苷酶转化柚皮苷生成普鲁宁的工艺,在此基础上分离纯化得到浓度较高的普鲁宁。利用碱性缓冲液处理柚苷酶,使其β-葡萄糖苷酶的活力大部分丧失,而保留α-鼠李糖苷酶的活性,从而使酶水解柚皮苷的产物——普鲁宁得到积累。利用水解过程中还原糖生成量与普鲁宁生成量成正比的关系,用DNS法检测柚皮苷水解过程中还原糖的含量,分析不同工艺参数中普鲁宁的含量。通过对柚皮苷水解过程的动态分析,得到柚苷酶转化柚皮苷的最适条件:温度50℃,p H 4.0,酶用量10 U/m L,底物含量0.16%。在此条件下酶解90 min,普鲁宁还保持较高的浓度而柚皮苷基本转化完全。酶解液经化学萃取和硅胶柱层析,得到纯化的普鲁宁,用高效液相色谱检测,其纯度达到99%。This paper is mainly involved in the process optimization of prunin production from naringin catalyzed with naringinase, and subsequent prunin purification. Firstly, naringinase was treated with p H 12 of Na2HPO4/ Na OH buffer to deprive β-glucosidase activity but retain α-rhamnosidase activity. After treated by alkaline buffer, prunin could be accumulated during the process of naringin hydrolysis. Secondly, based on the proportional relationship between reducing sugar accumlation and prunin production, the naringin hydrolysis process was analyzed and optimized by means of detecting reducing sugar concentration with DNS method. The optimal condition for naringin bioconversion was determined as enzyme dosage 10 U/m L, substrate concentration 0.16%, p H 4.0, temperature 50 ℃. Under this optimal condition,prunin could remain much higher concentration than naringenin and naringin was almost converted completely at 90 min.Finally, the final product, obtained through chemic extraction and sillica gel column chromatography, was prunin identified by HPLC method, and the purity of prunin reached 99%.

关 键 词：柚皮苷 普鲁宁 柚苷酶 生物转化 

分 类 号：TQ464[化学工程—制药化工]