食品科学 ›› 2023, Vol. 44 ›› Issue (8): 152-161.doi: 10.7506/spkx1002-6630-20220427-349

• 生物工程 • 上一篇    下一篇

蛹虫草产β-葡萄糖苷酶与α-L-阿拉伯呋喃糖苷酶活性分析及转化人参皂苷Rg1与Rc应用

李飞, 黄秋婷, 隋新, 谢莹   

  1. (1.吉林化工学院生物与食品工程学院,吉林 吉林 132022;2.江南大学生物技术学院,工业生物技术教育部重点实验室,江苏 无锡 214122)
  • 出版日期:2023-04-25 发布日期:2023-05-06
  • 基金资助:
    吉林省教育厅科学技术研究规划项目(JJKH20220247KJ);吉林市科技创新创业人才培育计划项目(20213824); 吉林化工学院重大科研项目(2020016)

Activities of β-Glucosidase and α-L-Arabinofuranosidase from Cordyceps militaris and Their Applications in the Transformation of Ginsenoside Rg1 and Rc

LI Fei, HUANG Qiuting, SUI Xin, XIE Ying   

  1. (1. School of Biological and Food Engineering, Jilin Institute of Chemical Technology, Jilin 132022, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China)
  • Online:2023-04-25 Published:2023-05-06

摘要: 通过鉴定筛选得到1 株高产β-葡萄糖苷酶的长白山虫草属真菌蛹虫草。研究碳源、氮源及pH值对菌株产β-葡萄糖苷酶、α-L-阿拉伯呋喃糖苷酶、虫草酸与生物量的影响规律,从而获得高产生物活性物质的培养条件,并对蛹虫草转化人参皂苷Rg1与Rc的路径与转化率进行了研究。采用紫外检测法测定β-葡萄糖苷酶与α-L-阿拉伯呋喃糖苷酶活力,超高效液相色谱-四极杆飞行时间质谱法鉴定转化产物中人参皂苷的组成,利用高效液相色谱法测定虫草素含量及人参皂苷的转化率。结果表明:蛹虫草在碳源为纤维二糖、氮源为牛肉膏、pH 8、培养120 h条件下有较高β-葡萄糖苷酶活力((74.70±0.09)U/mL)。在碳源为乳糖、氮源为蛋白胨、pH 4、培养72 h条件下有最高的α-L-阿拉伯呋喃糖苷酶活力((11.55±0.01)U/mL)。蛹虫草转化人参皂苷Rg1的路径为Rg1→Rh1和Rg1→F1,转化Rc路径为Rc→Rd→Rg3→CK和Rc→CMc。经过168 h的转化,人参皂苷Rg1转化率为54.9%,Rc转化率达到83.44%。本研究为提高药食两用真菌蛹虫草生物转化人参皂苷效率提供了理论基础,也为蛹虫草和人参食品、药品开发提供了一定理论支持。

关键词: 蛹虫草;β-葡萄糖苷酶;α-L-阿拉伯呋喃糖苷酶;人参皂苷

Abstract: In this study, a strain of Cordyceps militaris with high productivity of β-glucosidase was identified and selected. The effects of carbon source, nitrogen source and pH value on the production of β-glucosidase, α-L-arabinofuranosidase, cordyceps acid and biomass by the strain were studied to determine the optimal culture conditions for the production of bioactive substances with high productivity. The transformation pathways and efficiency of ginsenosides Rg1 and Rc by C. militaris were evaluated. The activities of β-glucosidase and α-L-arabinofuranidase were determined by ultraviolet (UV) spectrophotometry. The ginsenoside composition of the transformed products was identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and the content of cordycepin and the transformation efficiency of ginsenoside were determined by high performance liquid chromatography (HPLC). The results showed that the highest β-glucosidase activity of ((74.70 ± 0.09) U/mL) was obtained after 120 h culture using cellulose disaccharide as the carbon source and beef extract as the nitrogen source at pH 8, while the highest α-L-arabinofuranosidase activity of ((11.55 ± 0.01) U/mL) was obtained after 72 h culture using lactose as the carbon source and peptone as the nitrogen source at pH 4. The transformation pathways of ginsenoside Rg1 by C. militaris were Rg1→Rh1 and Rg1→F1, and the transformation pathways of Rc were Rc→Rd→Rg3→CK and Rc→CMc. The conversion efficiency of ginsenoside Rg1 and Rc was 54.9% and 83.44% after 168 h, respectively. This study provides a theoretical basis to improve the efficiency of biotransformation of ginsenoside by the medicinal and edible fungus C. militaris, and lays theoretical support for developing food and medicinal products from C. militaris and ginseng.

Key words: Cordyceps militaris; β-glucosidase; α-L-arabinofuranosidase; ginsenoside

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