Biomacromolecules, the first components of bioactive substances in traditional Chinese medicines (TCM) have wide bioactivity-related efficacy but have not yet been fully appreciated compared to small molecule components. The present review brings a novel and systemic point of view to deepen the understanding of the pharmacodynamic material basis of TCM based on biomacromolecules (polysaccharides, proteins and nucleic acids). Biomacromolecules have been, are and will have considerable roles in the efficacy of Chinese medicine, as evidenced by the number of biological activities related to traditional clinical efficacy. The direct and indirect mechanisms of biomacromolecules are further accounted for in a variety of neurotransmitters, hormones, and immune substances to maintain immune function in both sensitive and stable equilibrium. The biological functions of biomacromolecules have been elaborated on in regard to their roles in the process of plant growth and development to the relationship between primary metabolism and secondary metabolism and to the indispensable role of polysaccharides, proteins, and nucleic acids in the quality formation of TCM. Understanding the functional properties and mechanisms of biological macromolecules will help to demystify the drug properties and health benefits of TCM.
Biphenyls are unique phytoalexins de novo synthesized in plants in response to pathogen attack. These compounds are found in Maloideae, a subfamily of the Rosaceae. The anti-microbial activities of biphenyls have been reported in a number of studies and they appear to represent an important defense strategy against pathogens common in the Maloideae, such as species in Malus, Pyrus, Sorbus, and Chaenomeles. Here, cell suspension cultures of Sorbus pohuashanensis were established to study biphenyl phytoalexins formation after yeast extract (YE) treatment. An ultra-performance liquid chromatography (UPLC) method coupled with quadrupole time of flight mass spectrometry (Q-TOF-MS) LC−MS/MS was applied to determine the time course of these biphenyl biomarkers accumulation in YE-treated S. pohuashanensis suspension cells. The results of quantitative analyses show the content of Noraucuparin, 2 -Hydroxyaucuparin, and their glycosides initially increased, then decreased over time. The Noraucuparin content reached its highest (225.76 µg·g −1 ) at 18 h after treatment, 6 hours earlier than that of Noraucuparin 5-O-β-D-glucopyranoside. The content of 2 -Hydroxyaucuparin reached its highest (422.75 µg·g −1 ) at 30 h after treatment, also earlier than that of its glycoside. The understanding of phytoalexin metabolism in this study may provide a basis for improving Maloideae resistance to pathogens.
Shikonin and its derivatives are the main active components in the medicinal plant Arnebia euchroma and possess extensive pharmaceutical properties. In this study, we developed an optimized yeast system to obtain high-level production of 3-geranyl-4-hydroxybenzoate acid (GBA), an important intermediate involved in shikonin biosynthesis pathway. For host selection, recombinant expression of p-hydroxybenzoate:geranyltransferase (PGT) derived from A. euchroma was performed in Saccharomyces cerevisiae WAT11U strain and high yield monoterpene strain. In shake flask culture with 1 mM p-hydroxybenzoate acid (PHBA), they could yield GBA at 0.1567 and 20.8624 mg L-1, respectively. Additionally, AePGT6 showed higher enzymatic activity than its homologs. Moreover, by combining improvement in the homologous mevalonate pathway with reconstruction in the heterologous shikimic pathway, a de novo GBA synthesis pathway was constructed in StHP6tHC with co-overexpressed SctHMG1, optimized EcUbiC and AePGT6. A high titer of 179.29 mg L-1 GBA was achieved in StHP6tHC under shake flask fermentation with 1 mM PHBA. These results suggest that yeast could be engineered systematically to enable an efficient monoterpene-quinone or naphthoquinone production.
Achyranthes bidentata Blume (AB) is a health food and a sulfur-free herbal medicine that is one of the most heavily sulfur-fumigated herbs in the marketplace. In this work, a comprehensive approach using ultra-performance liquid chromatography coupled with quadrupole time-of-flight-MS (UPLC-Q-TOF-MS) and multivariate statistical analysis was developed to identify characteristic sulfur-fumigation markers, elucidate chemical transformation mechanisms and characterize the degree of sulfur-fumigation of AB. Non-fumigated and sulfur-fumigated AB samples were compared by UPLC-Q-TOF-MS/MS analysis. Three triterpene saponins (Betavulgarosides II–IV) and two amides (Feruloyl-4-O-methyldopamine and Moupinamide) were identified as characteristic markers, which were positively correlated with two active AB components, namely oleanic acid and ferulic acid, respectively. Moreover, the extent of the sulfur-fumigation under different weight ratios of sulfur to herbal materials (1:20, 1:40, and 1:80) was analyzed based on chemical transformations and sulfur dioxide residues. Further verification showed that the ratio of 1:40 within 1 h was reasonable and efficient for herb quality preservation and assurance. This study provides a reliable sulfur-fumigation protocol for the quality control of AB and other herbs.
Root rot disease caused by Fusarium oxysporum is a devastating disease of Salvia miltiorrhiza and dramatically affected the production and quality of Sa. miltiorrhiza. Besides the agricultural and chemical control, biocontrol agents can be utilized as an additional solution. In the present study, an actinomycete that highly inhibited F. oxysporum was isolated from rhizosphere soil and identified as based on morphological and molecular characteristics. Greenhouse assay proved that the strain had significant biological control effect against Sa. miltiorrhiza root rot disease and growth-promoting properties on Sa. miltiorrhiza seedlings. To elucidate the biocontrol and plant growth-promoting properties of St-220, we employed an analysis combining genome mining and metabolites detection. Our analyses based on genome sequence and bioassays revealed that the inhibitory activity of St-220 against F. oxysporum was associated with the production of enzymes targeting fungal cell wall and metabolites with antifungal activities. Strain St-220 possesses phosphate solubilization activity, nitrogen fixation activity, siderophore and indole-3-acetic acid production activity in vitro, which may promote the growth of Sa. miltiorrhiza seedlings. These results suggest that St. albidoflavus St-220 is a promising biocontrol agent and also a biofertilizer that could be used in the production of Sa. miltiorrhiza.
Trichosanthis Radix (TR) is one of the most severely sulfur-fumigated herbs in the market, whose transformation mechanism of chemical compositions and sulfur-fumigation markers of TR have not been clarified. To excavate characteristic sulfur-fumigation markers of TR samples, this study brings up a practical protocol using both ultraperformance liquid chromatography/quadrupole time-of-flight-mass spectrum (UPLC-ESI-QTOF-MS/MS)-based non-targeted metabolomics and ultra-performance liquid chromatography/electrospray ionization/quadrupole multiple-stage linear ion-trap mass spectrum (UPLC-ESI-QTRAP-MS/MS)-based widely targeted metabolomics. The results of study demonstrated that five characteristic markers are sulfur-containing components, which were identified as p-Hydroxybenzyl hydrogen sulfite, cucurbitacin D sulfite I, cucurbitacin D sulfite II, cucurbitacin B sulfite I, and cucurbitacin B sulfite II, respectively. Additionally, cucurbitacin B and D were also filtered and identified as the characteristic sulfur-fumigation markers. Meanwhile, the different sulfur-fumigation extent of TR samples was tested by chemical transformations analysis and sulfur dioxide residues test. Further, 58.16% (139 of 239) of the differential metabolites content significantly reduced in sulfur-fumigated TR samples. Besides, 20 kinds of non-sulfur marker metabolites were tested to evaluate the quality of TR samples before and after sulfur fumigation, predominantly including phenolic acids, amino acids, lipids and nucleotides. Taking TR as an example, this work provides a comprehensive practical protocol for the quality supervision of sulfur-fumigation herbs.
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