Efficient enzymatic hydrolysis of Protogracillin for clean preparation of Prosapogenin A by response surface methodology optimization
Efficient enzymatic hydrolysis of Protogracillin for clean preparation of Prosapogenin A by response surface methodology optimization
复制标题
通过响应面法优化，高效酶解原细青霉素清洁制备原皂苷元 A
DOI：
10.1080/17518253.2022.2138723
复制
发表时间：
2022-07-03
影响因子：
6.6
通讯作者：
Yang，Huan
中科院分区：
文献类型：
--
作者：
Xie，Haicheng;Zhang，Le;Yang，Huan
原阔叶皂苷A（也称为原阔叶皂苷III和多叶根素V）是薯蓣块茎中的一种次生甾体皂苷，具有比原甲氨酰胺更优越的药理活性。传统的制备方法，如柱层析、酸水解等，由于步骤繁琐、有机溶剂消耗大，效率很低，规模有限。本研究旨在建立一种简便、高效的酶法水解原料中丰富的原格拉西林获得原apogenin a的方法。考虑到最高的水解性能，我们从四种商业酶中选择了β-葡聚糖酶。采用Box-Behnken设计优化响应面法，在含有β-葡聚糖酶/原氨苄青霉素（5.0:1,w/w）的0.20 M HAc-NaAc缓冲液（pH 4.81）中进行酶解，在56.7°C水浴中持续保持4 h。结果表明，原格拉西林几乎完全水解为促apogenin A，产率最高为96.4±1.4%。该方法高效、便捷地获得促apogenin A，旨在为促apogenin A的工业化制备提供实验室规模实验基础。图形抽象
ABSTRACT Prosapogenin A (also known as Progenin III and polyphyllin V), a secondary steroidal saponin in Dioscorea zingiberensis tubers, has more superior pharmacological activities than its primary form Protogracillin. Conventional preparation methods, such as column chromatography, and acidic hydrolysis, are of very low efficiency and limited scale due to tedious procedures and large consumption of organic solvent. This study aims to establish a convenient method for efficiently obtaining Prosapogenin A by enzymatic hydrolysis of abundant Protogracillin in the raw material. In light of the highest hydrolysis performance, β-dextranase was selected from four commercial enzymes in this application. After optimization of the conditions by the response surface methodology using Box–Behnken design, the enzymatic hydrolysis was carried out in 0.20 M HAc-NaAc buffer (pH 4.81) containing β-dextranase/Protogracillin (5.0:1, w/w), and the system was constantly kept at 56.7°C water bath for 4 h. Consequently, Protogracillin has been almost completely hydrolyzed to be Prosapogenin A and the highest yield was 96.4 ± 1.4%. The newly proposed approach is efficient and promising for conveniently obtaining Prosapogenin A and aimed to provide a laboratory-scale experimental foundation for the preparation of Prosapogenin A in industrial applications. GRAPHICAL ABSTRACT