Department of Pharmaceutical Botany, Faculty of Pharmacy, Jagiellonian University Medical College, 30-688 Krakow, Poland.
SSG of Medicinal Plants and Mushroom Biotechnology Department of Pharmaceutical Botany, Jagiellonian University Medical College, Medyczna 9 Str., 30-688 Cracow, Poland.
Molecules. 2021 Aug 17;26(16):4965. doi: 10.3390/molecules26164965.
The aims of this study were to evaluate the antioxidant properties, to investigate the content of major secondary metabolites in cell cultures, and to determine the change in the production of phenolic acids by adding phenylalanine to the culture medium. Three in vitro methods, which depend on different mechanisms, were used for assessing the antioxidant activity of the extract: 1,1-diphenyl-2-picrylhydrazil (DPPH), reducing power and Fe chelating activity assays. The extract showed moderate activity both in the DPPH and in the reducing power assays (IC = 1.966 ± 0.058 mg/mL; ASE/mL = 16.31 ± 1.20); instead, it was found to possess good chelating properties reaching approximately 70% activity at the highest tested dose. The total phenolic, total flavonoid, and condensed tannin content of cell culture extract was spectrophotometrically determined. The phenolic acid content was investigated by RP-HPLC, and the major metabolites-protocatechuic and -hydroxybenzoic acids-were isolated and investigated by H NMR. The results showed that phenylalanine added to cell cultures at concentrations of 100, 150, and 200 mg/150 mL increased the production of phenolic acids. Cultures that were grown for 3 weeks and collected after 4 days of phenylalanine supplementation at high concentration showed maximal content of phenolic acids (73.76 mg/100 g DW).
本研究旨在评估抗氧化性能，研究细胞培养物中主要次生代谢物的含量，并通过向培养基中添加苯丙氨酸来确定酚酸产量的变化。三种体外方法，它们依赖于不同的机制，用于评估提取物的抗氧化活性：1,1-二苯基-2-苦基肼基（DPPH）、还原力和 Fe 螯合活性测定。提取物在 DPPH 和还原力测定中均表现出中等活性（IC = 1.966 ± 0.058 mg/mL；ASE/mL = 16.31 ± 1.20）；然而，它被发现具有良好的螯合性能，在最高测试剂量下达到约 70%的活性。细胞培养物提取物的总酚、总黄酮和缩合单宁含量通过分光光度法测定。通过 RP-HPLC 研究了酚酸含量，并通过 1H NMR 分离和研究了主要代谢物-原儿茶酸和 -羟基苯甲酸。结果表明，苯丙氨酸以 100、150 和 200 mg/150 mL 的浓度添加到细胞培养物中，增加了酚酸的产量。在高浓度苯丙氨酸添加 4 天后培养 3 周并收集的培养物显示出最大的酚酸含量（73.76 mg/100 g DW）。
