Kuo Chia-Hung, Liu Hsuan-Chieh, Nargotra Parushi, Chan Hsiao-Sung, Shih Hsin-Der, Liu Yung-Chuan
Department of Seafood Science, National Kaohsiung University of Science and Technology, Kaohsiung 811, Taiwan.
Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan.
Molecules. 2025 Apr 21;30(8):1859. doi: 10.3390/molecules30081859.
S. MOORE, a medicinal plant, is recognized for tetrandrine production, which is extensively accepted for its therapeutic benefits. However, the slow growth of limits tetrandrine's availability, which can be addressed by in vitro hairy root cultivation using and optimization of medium components. The present study attempted the three-step optimization of three components of woody plant medium (ammonium nitrate, calcium nitrate, and sucrose), including two-level factorial design, path of steepest ascent (PSA), and central composite design (CCD) to obtain high hairy root biomass and tetrandrine production. The CCD-based optimization for enhanced hairy root biomass resulted in a hairy root biomass of 9.75 g dw/L at optimal concentrations of ammonium nitrate (NHNO), calcium nitrate (Ca(NO)), and sucrose of 631.96 mg/L, 651 mg/L, and 41.35 g/L, respectively. The optimal concentration of 550.31 mg/L, 862.88 mg/L, and 25.89 g/L for NHNO, Ca(NO), and sucrose, respectively, obtained after CCD analysis for enhanced tetrandrine production led to the maximum tetrandrine of 70.48 mg/L. Medium optimization resulted in a 1.47-fold increase in hairy root biomass and a 1.37-fold increase in tetrandrine production under individually optimized conditions. The present study findings confirmed the important role of process optimization for enhanced product yield.
千金藤是一种药用植物,因其能产生粉防己碱而闻名,粉防己碱因其治疗功效而被广泛认可。然而,千金藤生长缓慢限制了粉防己碱的产量,这可以通过使用发根农杆菌进行体外毛状根培养和优化培养基成分来解决。本研究尝试对木本植物培养基的三种成分(硝酸铵、硝酸钙和蔗糖)进行三步优化,包括两水平析因设计、最速上升路径(PSA)和中心复合设计(CCD),以获得高毛状根生物量和粉防己碱产量。基于CCD的提高毛状根生物量的优化在硝酸铵(NH₄NO₃)、硝酸钙(Ca(NO₃)₂)和蔗糖的最佳浓度分别为631.96 mg/L、651 mg/L和41.35 g/L时,得到了9.75 g dw/L的毛状根生物量。在基于CCD分析提高粉防己碱产量后,分别获得的硝酸铵、硝酸钙和蔗糖的最佳浓度为550.31 mg/L、862.88 mg/L和25.89 g/L,导致最大粉防己碱产量为70.48 mg/L。培养基优化在单独优化的条件下使毛状根生物量增加了1.47倍,粉防己碱产量增加了1.37倍。本研究结果证实了工艺优化对提高产品产量的重要作用。