Skrzypczak-Pietraszek Ewa, Urbańska Aneta, Żmudzki Paweł, Pietraszek Jacek
Chair and Department of Pharmaceutical Botany, Collegium Medicum Jagiellonian University Kraków Poland.
Chair of Pharmaceutical Chemistry, Faculty of Pharmacy, Collegium Medicum Jagiellonian University Kraków Poland.
Eng Life Sci. 2019 Aug 6;19(12):931-943. doi: 10.1002/elsc.201900051. eCollection 2019 Dec.
(L.) Urban is an important pharmacopoeial plant used not only in medicine but also in cosmetology. agitated shoot cultures were established to study the influence of ethephon, methyl jasmonate, l-phenylalanine (Eth 50 µM, MeJa 50 µM, L-Phe 2.4 g/L of medium, respectively; seven variants of the supplementation) on the accumulation of secondary metabolites: the main centellosides (asiaticoside and madecassoside) and selected phenolic acids, and flavonoids in the biomass. Microshoots were harvested two and six days after the supplementation. Secondary metabolites were analyzed in methanolic extracts by UPLC-MS/MS (centellosides) and by HPLC-DAD (phenolics). In comparison with the reference cultures, the concentrations of individual secondary metabolites increased as follows: centellosides up to 5.6-fold (asiaticoside), phenolic acids up to 122-fold (-coumaric acid) and flavonoids up to 22.4-fold (kaempherol). The highest production increase of individual compounds was observed for different variants of supplementation. Variant C (50 µM MeJa), the most optimal for centellosides and flavonoid accumulation, was selected for the experiment with bioreactors. Bioreactor Plantform™, compared to RITA system and agitated cultures, appeared to be the most advantageous for secondary metabolites production in shoot cultures. The phenolic acid, flavonoid, centelloside, and total secondary metabolite productivity in Plantform™ system is 1.8-fold, 1.7-fold, 2.8-fold, 2.1-fold, respectively, higher than in MeJa elicitated agitated cultures, and 4.3-fold, 7.3-fold, 12.2-fold, 7.2-fold, respectively, higher than in control agitated cultures.
(L.) 积雪草是一种重要的药典植物，不仅用于医学，还用于美容学。建立了振荡培养的芽培养物，以研究乙烯利、茉莉酸甲酯、L-苯丙氨酸（分别为50 μM乙烯利、50 μM茉莉酸甲酯、2.4 g/L L-苯丙氨酸于培养基中；七种添加变体）对次生代谢产物积累的影响：主要的积雪草苷（积雪草苷和羟基积雪草苷）以及选定的酚酸和黄酮类化合物在生物量中的积累。在添加后两天和六天收获微芽。通过超高效液相色谱-串联质谱法（积雪草苷）和高效液相色谱-二极管阵列检测法（酚类化合物）分析甲醇提取物中的次生代谢产物。与对照培养物相比，各个次生代谢产物的浓度增加如下：积雪草苷高达5.6倍（积雪草苷）、酚酸高达122倍（对香豆酸）和黄酮类化合物高达22.4倍（山奈酚）。不同添加变体的单个化合物产量增加最高。选择变体C（50 μM茉莉酸甲酯）进行生物反应器实验，其对积雪草苷和黄酮类化合物积累最为理想。与RITA系统和振荡培养相比，Bioreactor Plantform™似乎对芽培养物中次生代谢产物的生产最为有利。Plantform™系统中酚酸、黄酮类化合物、积雪草苷和总次生代谢产物的生产率分别比茉莉酸甲酯诱导的振荡培养高1.8倍、1.7倍、2.8倍、2.1倍，比对照振荡培养分别高4.3倍、7.3倍、12.2倍、7.2倍。
