二氢去氢双松柏醇的微生物转化研究

Study on Microbial Transformation of Dihydrodehydrodiconiferyl Alcohol

利用微生物转化的方法,以Bacillus subtilisYD 016为转化菌株,对二氢去氢双松柏醇的转化进行研究。该菌株可以将二氢去氢双松柏醇完全转化,主要得到3种转化产物(记为HMZG-1、HMZG-2、HZMG-3)。通过硅胶柱层析和制备型HPLC方法对转化产物进行分离纯化,经ESI-MS、~1H-NMR和^(13)C-NMR分析,转化产物分别鉴定为二氢去氢双松柏醇-4-O-β-D-吡喃葡萄糖苷(HMZG-1)、二氢去氢双松柏醇-9'-O-β-D-吡喃葡萄糖苷(HMZG-2)、二氢去氢双松柏醇-9-O-β-D-吡喃葡萄糖苷(HMZG-3)。通过高效液相色谱法对生物转化情况进行检测,建立转化产物和底物的标准曲线。根据HPLC结果计算可知,3种转化产物的转化率分别为:17.34%,14.29%,66.41%。该研究丰富了Bacillus subtilis YD 016的底物作用范围,为二氢去氢双松柏醇葡萄糖苷的合成提供了一条新途径。

In this research,the microbial transformation of dihydrodehydrodiconiferyl alcohol was studied. Through strain screening,Bacillus subtilis YD 016 was chosen to biotransform dihydrodehydrodiconiferyl alcohol. By two-stage culture method,dihydrodehydrodiconiferyl alcohol was biotransformed into three metabolites completely（ denoted as HMZG-1,HMZG-2 and HMZG-3） by Bacillus subtilis YD 016. Then silica gel column chromatography and preparative high performance liquid chromatography was performed to isolate and purify the transformation products. And the structures of three metabolites were elucidated by spectroscopic method.According to the Electrospray Ionization Mass Spectroscopy,1H-NMR and13C-NMR spectroscopy analysis data,which are consistent with those of the literatures,three transformation products were identified as dihydrodehydrodiconiferyl alcohol 4-O-β-D glucopyranoside（ HMZG-1）,dihydrodehydrodiconiferyl alcohol 9＇-O-β-D glucopyranoside（ HMZG-2）,dihydrodehydrodiconiferyl alcohol 9-O-β-D glucopyranoside（ HMZG-3） separately. High performance liquid chromatography spectrum method was established to detect biotransformation substrate and products in order to quantify their contents. By analysing the results of HPLC spectrum data,the converting rate of three transformation products was determined as 17. 34%,14. 29% and 66. 41% respectively. Time course experiments detected that dihydrodehydrodiconiferyl alcohol could be metabolized into dihydrodehydrodiconiferyl alcohol β-D-glucopyranosides within 72 h,so 72 h was chosen as final conversion termination time. This study provides a new approach to synthesize dehydrodiconiferyl alcohol β-D-glucopyranoside and enriches the range of substrate Bacillus subtilis YD 016 catalysis.