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菌株 XD101 对人参皂苷 Rb1 的生物转化为人参皂苷 CK:一种安全的生物转化策略。

Biotransformation of Ginsenoside Rb1 to Ginsenoside CK by Strain XD101: a Safe Bioconversion Strategy.

机构信息

Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Taibai North Road 229, Xi'an, 710069, China.

Shaanxi R&D Center of Biomaterials and Fermentation Engineering, School of Chemical Engineering, Northwest University, Xi'an, 710069, China.

出版信息

Appl Biochem Biotechnol. 2021 Jul;193(7):2110-2127. doi: 10.1007/s12010-021-03485-0. Epub 2021 Feb 24.

Abstract

Ginsenoside Rb1 is the main predominant component in Panax species. In this study, an eco-friendly and convenient preparation method for ginsenoside CK has been established, and five strains of β-glucosidase-producing microorganisms were screened out from the soil of a Panax notoginseng planting field using Esculin-R2A agar. Aspergillus niger XD101 showed that it has excellent biocatalytic activity for ginsenosides; one of the isolates can convert ginsenoside Rb1 to CK using extracellular enzyme from the mycelium. Mycelia of A. niger were cultivated in wheat bran media at 30 °C for 11 days. By the removal of mycelia from cultured broth, enzyme salt fractionation by ammonium sulfate (70%, v/v) precipitation, and dialysis, sequentially, crude enzyme preparations from fermentation liquid supernatant were obtained. The enzymatic transformed Rb1 as the following pathways: Rb1→Rd→F2→CK. The optimized reaction conditions are at reaction time of 72 h, in the range of pH 4-5, and temperature of 50-60 °C. Active minor ginsenosides can be obtained by a specific bioconversion via A. niger XD101 producing the ginsenoside-hydrolyzing β-glucosidase. In addition, the crude enzyme can be resulted in producing ginsenoside CK via conversion of ginsenoside Rb1 at high conversion yield (94.4%). FDA generally regarded, A.niger as safe microorganism. Therefore, these results indicate that A. niger XD10 may provide an alternative method to prepare ginsenoside CK without food safety issues in the pharmaceutical industry.

摘要

人参皂苷 Rb1 是人参属植物中的主要优势成分。本研究建立了一种绿色、简便的制备人参皂苷 CK 的方法,从三七种植地土壤中筛选出 5 株产β-葡萄糖苷酶的微生物,采用 Esculin-R2A 琼脂进行筛选。黑曲霉 XD101 对人参皂苷表现出优异的生物催化活性;其中一株菌可以通过胞外酶将人参皂苷 Rb1 转化为 CK。在 30℃下用麦麸培养基培养黑曲霉 XD101 菌丝体 11 天。通过从培养物中去除菌丝体,依次用硫酸铵(70%,v/v)沉淀和透析进行酶盐分级,从发酵液上清液中获得粗酶制剂。酶转化 Rb1 的途径如下:Rb1→Rd→F2→CK。优化的反应条件为反应时间 72 h,pH4-5,温度 50-60℃。通过黑曲霉 XD101 产生的人参皂苷水解β-葡萄糖苷酶的特异性生物转化,可以获得活性较低的人参皂苷。此外,粗酶制剂可通过转化人参皂苷 Rb1 生成人参皂苷 CK,转化率高达 94.4%。FDA 通常认为黑曲霉是安全的微生物。因此,这些结果表明,黑曲霉 XD10 可能为制药工业中制备人参皂苷 CK 提供一种无食品安全问题的替代方法。

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