Ultrahigh diterpenoid tanshinone production through repeated osmotic stress and elicitor stimulation in fed-batch culture of Salvia miltiorrhiza hairy roots.
Hyperosmotic stress (OS, created with 50 g/L sorbitol) and a yeast elicitor (YE, polysaccharide fraction of yeast extract) applied to Salvia miltiorrhiza hairy root cultures had a synergistic effect on the diterpenoid tanshinone production. With a single OS+YE treatment and nutrient feeding, the total tanshinone content of roots was increased by sevenfold (from 0.2 to 1.6 mg/g dry weight (dw)) and the volumetric yield by 13-fold (from 1.95 to 27.4 mg/L) compared to the batch control culture. With repeated feeding of OS and nutrient medium in an extended fed-batch culture process (i.e., 10 mL fresh medium with 50 g/L sorbitol 25 mg/L YE, every 5 days from day 21 to day 60), the total tanshinone content of roots was increased to 18.1 mg/g dw (or 1.8 wt.%) and the volumetric tanshinone yield to 145 mg/L, which were about 100-fold and 70-fold of those, respectively, in the batch control. Another interesting finding was the presence of root fragments (fine particles) with extremely high tanshinone content in the OS+YE treated cultures. It was also possible to reuse the sorbitol medium for the hairy root growth and tanshinone production to reduce the medium expenses.
Wu
Jian-Yong
JY
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong. bcjywu@polyu.edu.hk
Shi
Ming
M
D045784:Abietanes; D018533:Biomass; D019149:Bioreactors; D003470:Culture Media / Q000737:chemistry / Q000191:economics; D004224:Diterpenes / Q000378:metabolism; D009997:Osmotic Pressure; D010616:Phenanthrenes / Q000378:metabolism; D018517:Plant Roots / Q000254:growth & development / Q000378:metabolism*; D010946:Plants, Medicinal / Q000254:growth & development / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000494:pharmacology; D027561:Salvia miltiorrhiza / Q000378:metabolism*; D046509:Tissue Culture Techniques / Q000379:methods*
D045784:Abietanes; D003470:Culture Media; D004224:Diterpenes; D010616:Phenanthrenes; D011135:Polysaccharides, Bacterial; C021751:tanshinone