化工学报 ›› 2014, Vol. 65 ›› Issue (9): 3583-3591.DOI: 10.3969/j.issn.0438-1157.2014.09.037

• 生物化学工程与技术 • 上一篇    下一篇

两相体系中β-葡萄糖苷酶催化栀子苷水解制备京尼平

万丽花1, 姚忠1, 倪芳1, 韦敏1, 周治1, 王浩绮2, 孙芸1, 仲兆祥1   

  1. 1 南京工业大学食品与轻工学院, 江苏 南京 211816;
    2 南京工业大学生物与制药工程学院, 江苏 南京 211816
  • 收稿日期:2014-01-08 修回日期:2014-05-22 出版日期:2014-09-05 发布日期:2014-09-05
  • 通讯作者: 仲兆祥
  • 基金资助:

    国家自然科学基金项目(21206072);江苏省农业支撑计划项目(BE2012399);江苏省自然科学基金项目(BK2012825);国家科技支撑计划项目(2012BAD34B08)。

Biosynthesis of genipin from gardenoside catalyzed by β-glucosidase in two-phase medium

WAN Lihua1, YAO Zhong1, NI Fang1, WEI Min1, ZHOU Zhi1, WANG Haoqi2, SUN Yun1, ZHONG Zhaoxiang1   

  1. 1 School of Food Science and Light Industry, Nanjing University of Technology, Nanjing 211816, Jiangsu, China;
    2 School of Biological and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 211816, Jiangsu, China
  • Received:2014-01-08 Revised:2014-05-22 Online:2014-09-05 Published:2014-09-05
  • Supported by:

    supported by the National Natural Science Foundation of China (21206072), Agricultural Support Project in Jiangsu Province (BE2012399), Natural Science Foundation of Jiangsu Province (BK2012825), National Science and Technology Support Program (2012BAD34B08).

摘要: 京尼平是一种天然生物交联剂,且具有多种生理活性。本文以β-葡萄糖苷酶催化栀子苷水解制备京尼平为目标体系,证明了京尼平对β-D-葡萄糖苷酶具有竞争性抑制作用;考察了β-D-葡萄糖苷酶在不同有机相-水体系中的稳定性,发现其在正辛醇-水、正己醇-水体系中的稳定性均较好;测定了栀子苷在正辛醇-水和正己醇-水体系中的分配系数kD,gardenoside分别为0.17和0.76,而京尼平的分配系数kD,genipin为42.57和37.75;分别在水、正辛醇-水和正己醇-水体系中进行栀子苷水解制备京尼平的反应,当底物浓度为0.25 μmol·ml-1时京尼平收率分别为89.17%、93.96%、90.16%;进一步提高底物浓度为2.0 μmol·ml-1时,正辛醇-水体系中京尼平收率高达91.9%,较水相体系(79.7%)提高了12.2%。本文的研究结果证明了采用正辛醇-水两相反应体系可部分解除产物抑制,提高产物收率,并简化后续的产物分离。

关键词: 栀子苷, β-葡萄糖苷酶, 京尼平, 两相体系, 催化

Abstract: Genipin is an important bio-crosslinking agent with multiple physiological activities. In this paper, the competitive inhibition for activity of β-glucosidase by genipin was demonstrated. The stabilities of β-glucosidase in different organic solvents were investigated. Stability of β-glucosidase was satisfactory in the octanol-water and hexanol-water medium. The partition coefficients for gardenoside in the octanol-water and hexanol-water system were 0.17 and 0.76, while those for genipin were 42.57 and 37.75 respectively. The process for preparation of genipin was carried out in water, octanol-water and hexanol-water respectively, while gardenoside concentration was 0.25 μmol·ml-1. The obtained genipin yields were 89.17%, 93.96% and 90.16% respectively. Furthermore, the effect of gardenoside concentration on genipin yield was also investigated. While in the octanol-water system, conversion rate of genipin was as high as 91.9% at the gardenoside concentration of 2.0 μmol·ml-1, an increase of 12.2% compared to that in water. Using the octanol-water medium was helpful for partially eliminating product inhibition, improving genipin yield and simplifying product recovery.

Key words: gardenoside, β-glucosidase, genipin, two phase medium, catalysis

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