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哈茨木霉CGMCC 2979生物转化栀子中的京尼平苷制备京尼平
引用本文:董悦生,刘乐平,包永明,郝爱鱼,秦莹,温祖佳,修志龙.哈茨木霉CGMCC 2979生物转化栀子中的京尼平苷制备京尼平[J].催化学报,2014,35(9):1534-1546.
作者姓名:董悦生  刘乐平  包永明  郝爱鱼  秦莹  温祖佳  修志龙
作者单位:a 大连理工大学生命科学与技术学院, 辽宁大连116024;
b 大连市药品检验所, 辽宁大连116021
基金项目:国家自然科学基金(81172966);家重点基础研究发展计划(973计划,2009CB724706);央高校基本科研业务费专项资金资助(DUT12ZD209)~~
摘    要:采用微生物直接转化药材的方法,将栀子中的京尼平苷转化为京尼平,无需糖苷酶和京尼平苷的制备. 在培养温度为30 ℃,pH 6.1以及栀子载量为80 g/L的条件下,48 h京尼平苷的转化率为97.8%. 转化后的京尼平通过XAD-16N大孔树脂偶联硅胶层析的方法,制备得到纯度大于95%的京尼平,收率为62.3%. 在催化、转化机制研究中,从哈茨木霉CGMCC2979的发酵液中分离得到了分子量为74.4 kDa的京尼平苷β-葡萄糖苷酶,该酶最优催化条件为50 ℃和pH 4.0-5.0. Km和Vmax分别为3.6 mmol/L和775 μmol/h/mg蛋白. 本文提供了一种简便、高效制备京尼平的新方法.

关 键 词:京尼平苷  京尼平  哈茨木霉  生物转化  京尼平苷β-葡萄糖苷酶  纯化
收稿时间:2014-02-14

Biotransformation of geniposide in Gardenia jasminoides to genipin by Trichoderma harzianum CGMCC 2979
Yuesheng Dong,Leping Liu,Yongming Bao,Aiyu Hao,Ying Qin,Zujia Wen,Zhilong Xiu.Biotransformation of geniposide in Gardenia jasminoides to genipin by Trichoderma harzianum CGMCC 2979[J].Chinese Journal of Catalysis,2014,35(9):1534-1546.
Authors:Yuesheng Dong  Leping Liu  Yongming Bao  Aiyu Hao  Ying Qin  Zujia Wen  Zhilong Xiu
Institution:a School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, Liaoning, China;
b Dalian Institute for Drug Control, Dalian 116021, Liaoning, China
Abstract:Trichoderma harzianum (T. harzianum) CGMCC 2979 was used to transform the geniposide in Gardenia jasminoides (G. jasminoides) to genipin, dispensing the use of purified enzyme and the extraction of geniposide from the raw material. At 30 ℃, pH 6.1, and an initial G. jasminoides concentration of 80 g dried fruit per liter of medium, the geniposide-to-genipin conversion rate reached 97.8% after 48 h of fermentation. The genipin was purified from the fermentation broth by a combined method of XAD-16N-resin and silica-gel chromatography, yielding a total recovery of 62.3%. A 74.4-kDa geniposide-β-glucosidase implicated in the transformation of geniposide to genipin was purified from T. harzianum CGMCC 2979. It had optimum activity at 50 ℃ and pH 4.0-5.0. The Km and Vmax of the enzyme for geniposide were 3.6 mmol/L and 775 μmol/h/mg protein, respectively. The simple, direct, and efficient biotransformation of geniposide in G. jasminoide to genipin by T. harzianum CGMCC 2979 that is described in this study could represent an alternative and effective method for producing genipin.
Keywords:Geniposide  Genipin  Trichoderma harzianum  Biotransformation  Geniposide-β-glucosidase  Purification
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