麦芽糖淀粉酶对葛根素进行体外酶促修饰生成葛根素糖苷

In vitro enzymatic modification of puerarin to puerarin glycosides by maltogenic amylase.

作者信息

Li Dan, Park Sung-Hoon, Shim Jae-Hoon, Lee Hee-Seob, Tang Shuang-Yan, Park Cheon-Seok, Park Kwan-Hwa

机构信息

National Laboratory for Functional Food Carbohydrate and Center for Agricultural Biomaterials, School of Agricultural Biotechnology, Seoul National University, Shillim-dong, Kwanak-gu, Seoul 151-742, Korea.

出版信息

Carbohydr Res. 2004 Dec 6;339(17):2789-97. doi: 10.1016/j.carres.2004.09.017.

DOI:10.1016/j.carres.2004.09.017

PMID:15542087

Abstract

Puerarin (daidzein 8-C-glucoside), the most abundant isoflavone in Puerariae radix, is prescribed to treat coronary heart disease, cardiac infarction, problems in ocular blood flow, sudden deafness, and alcoholism. However, puerarin cannot be given by injection due to its low solubility in water. To increase its solubility, puerarin was transglycosylated using various enzymes. Bacillus stearothermophilus maltogenic amylase (BSMA) was the most effective transferase used compared with Thermotoga maritima maltosyl transferase (TMMT), Thermus scotoductus 4-alpha-glucanotransferase (TS4alphaGTase), and Bacillus sp. I-5 cyclodextrin glucanotransferase (BSCGTase). TMMT and TS4alphaGTase lacked acceptor specificity for puerarin, which lacks an O-glucoside linkage between D-glucose and 7-OH-daidzein. The yield exceeded 70% when reacting 1% puerarin (acceptor), 3.0% soluble starch (donor), and 5U/100 microL BSMA at 55 degrees C for 45 min. The two major transfer products of the BSMA reaction were purified using C(18) and GPC chromatography. Their structures were identified as alpha-d-glucosyl-(1-->6)-puerarin and alpha-D-maltosyl-(1-->6)-puerarin using ESI+ TOF MS-MS and 13C NMR spectroscopy. The solubility of the transfer products was 14 and 168 times higher than that of puerarin, respectively.

摘要

葛根素（大豆苷元8 - C - 葡萄糖苷）是葛根中含量最为丰富的异黄酮，被用于治疗冠心病、心肌梗死、眼部血流问题、突发性耳聋和酒精中毒。然而，由于葛根素在水中的溶解度较低，无法进行注射给药。为提高其溶解度，利用多种酶对葛根素进行了转糖基化反应。与嗜热栖热菌麦芽糖基转移酶（TMMT）、嗜热栖热放线菌4-α-葡聚糖转移酶（TS4αGTase）和芽孢杆菌属I - 5环糊精葡聚糖转移酶（BSCGTase）相比，嗜热栖热芽孢杆菌麦芽糖生成淀粉酶（BSMA）是最有效的转移酶。TMMT和TS4αGTase对葛根素缺乏受体特异性，因为在D - 葡萄糖和7 - OH - 大豆苷元之间不存在O - 葡萄糖苷键。当1%的葛根素（受体）、3.0%的可溶性淀粉（供体）与5U/100μL的BSMA在55℃下反应45分钟时，产率超过70%。利用C(18)和凝胶渗透色谱法（GPC）对BSMA反应的两种主要转移产物进行了纯化。使用电喷雾电离正离子飞行时间串联质谱（ESI+ TOF MS - MS）和13C核磁共振光谱法（13C NMR）将它们的结构鉴定为α - d - 葡萄糖基 - (1→6) - 葛根素和α - D - 麦芽糖基 - (1→6) - 葛根素。转移产物的溶解度分别比葛根素高14倍和168倍。