Hsuan-Chieh Liu: Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan; Biomass Materials Technology Department, Agri-Industrial Systems Technology Division, Central Region Campus, Industrial Technology Research Institute, Nantou 54041, Taiwan. Electronic address: ohohpigjay@gmail.com.
Hsiao-Sung Chan: Department of Applied Chemistry, Chaoyang University of Technology, Taichung 413310, Taiwan. Electronic address: phyllanthus@gmail.com.
Parushi Nargotra: Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan. Electronic address: parushi11nargotra@gmail.com.
Hsin-Der Shih: Plant Pathology Division, Taiwan Agricultural Research Institute, Ministry of Agriculture, Taichung 413008, Taiwan. Electronic address: tedshih@tari.gov.tw.
Chia-Hung Kuo: Department of Seafood Science, National Kaohsiung University of Science and Technology, Kaohsiung 811, Taiwan; Center for Aquatic Products Inspection Service, National Kaohsiung University of Science and Technology, Kaohsiung 811, Taiwan. Electronic address: kuoch@nkust.edu.tw.
Yung-Chuan Liu: Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan. Electronic address: ycliu@dragon.nchu.edu.tw.
Tetrandrine, a bioactive active compound mainly found in the roots of Stephania tetrandra, exhibits various pharmacological properties. In vitro hairy root (HR) culture may serve as a promising solution for the extraction of tetrandrine, overcoming the limitations of natural cultivation. The present study describes the consistent production of tetrandrine from S. tetrandra hairy roots induced by different strains of Agrobacterium rhizogenes. Cultivation in woody plant medium (WPM) resulted in the highest HR biomass (0.056 g/petri-dish) and tetrandrine content (7.28 mg/L) as compared to other media. The maximum HR biomass (6.95 g dw/L) and tetrandrine production (68.69 mg/L) were obtained in the fifth week of cultivation. The presence of ammonium nitrate (800 mg/L), calcium nitrate (1156 mg/L), sucrose (20 g/L) and casein (2 g/L) enhanced the tetrandrine production. Moreover, the fed-batch cultivation demonstrated that the NHNO (1200 mg/L) was an important growth limiting factor that yielded the highest tetrandrine amount (119.59 mg/L). The cultivation of hairy roots in a mist trickling bioreactor for eight weeks was less (26.24 mg/L) than in the flask. Despite a lower tetrandrine yield observed in bioreactors compared to flask cultures, refining the growth medium and fine-tuning bioreactor operations hold promise for boosting tetrandrine yield.
