天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (11): 1928-1936.doi: 10.16333/j.1001-6880.2020.11.017

所属专题: No.8

• 开发研究 • 上一篇    下一篇

黄芩内生真菌发酵黄芩苷生成千层纸素A的代谢途径及转化条件研究

马宗敏,苗文丽,刘佳,何培,孙国强,裴林*   

  1. 河北省中医药科学院 浊毒证重点实验室,石家庄 050031

  • 出版日期:2020-11-28 发布日期:2020-12-07
  • 基金资助:
    国家重点研发计划(2017YFC1701701);河北省中医药管理局科研计划(2015087)

Study on pathway and fermentation conditions of an endophytic fungus transforming baicalin to oroxylin A

MA Zong-min,MIAO Wen-li,LIU Jia,HE Pei,SUN Guo-qiang,PEI Lin*#br#

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  1. Key Laboratory of Zhuodu Syndrome,Hebei Academy of Chinese Medical Sciences, Shijiazhuang 050031,China

  • Online:2020-11-28 Published:2020-12-07

摘要: 本研究采用黄芩内生青霉菌Penicillium sp.R3发酵转化黄芩苷生产千层纸素A,探索生物转化途径,优化发酵转化条件,旨在为天然产物千层纸素A的开发利用提供新方法、新途径。菌株Penicillium sp.R3在马铃薯葡萄糖(PDB)培养液中培养并进行转化反应,菌株生长4天后达到最大生物量,之后进入稳定期。通过不用底物添加法分析其生物转化途径,高效液相色谱检测发现菌株转化黄芩苷生成千层纸素A同时伴随黄芩素产生,且黄芩素为中间产物。单因素分析结合正交试验方法获取发酵转化的最适条件为:摇瓶装液量10%,接种孢子浓度1×106~2×106个/mL,30 ℃,150 rpm培养3天后添加0.08%黄芩苷转化6天。该条件下转化反应摩尔转化率达到42.44%,发酵液中千层纸素A产量达到177 mg/L,且结果稳定性好。实验明确了菌株生长及转化规律,获得了稳定且转化率较高的发酵转化条件,为千层纸素A的进一步开发利用奠定了基础。


关键词: 黄芩苷, 千层纸素A, 生物转化, 青霉菌R3

Abstract:

Aimed to provid a new approach for the exploitation of natural product oroxylin A,this reasearch focused on obtaining oroxylin A from baicalin by biotransformation applying endophytic fungus-Penicillium sp.R3 from Scutellaria baicalensis Georgi,identifying its biotransformation pathway and optimizing the fermentation conditions.The culture and biotransformation reaction were carried out in potato dextrose broth(PDB).Cell growth was measured by dry weight and the maximum biomass of Penicillium sp.R3 was gained on the day 4,then it entered the stationary phase.The biotransformation pathway was analyzed by the method of adding different substrates and the products were measured and identified by HPLC.After fermentation baicalin was converted to oroxylin A accompanied with baicalein which was identified as intermediate metabolite.Single factor analysis and orthogonal test were applied to optimized fermentation conditions.The optimal fermentation conditions were as follows:the liquid medium volume was 10% of the bottle;the concentration of inoculated spores was 1×106-2×106/mL and the temperature and shaker speed were 30 ℃,150 rpm respectively.The substrate baicalin was added on the day 3 with proportion of 0.08%,then the biotransformation process sustained 6 days.Under these optimal conditions the molar conversion rate and concentration of oroxylin A reached 42.44% and 177 mg/L respectively.The growth regularity and the biotransformation pathway of Penicillium sp.R3 was clarified,the stable and optimal conditions were obtained with a satisfactory yeild.All these findings lay a foundation for the further development and utilization of oroxylin A.

Key words: baicalin, oroxylin A, biotransformation, Penicillium , sp.R3

中图分类号:  R284.1 Q939.97