Ginsenosides are the primary bioactive components of ginseng, which is a popular medicinal plant that exhibits diverse pharmacological activities. Protopanaxadiol, protopanaxatriol and oleanolic acid are three basic aglycons of ginsenosides. Producing aglycons of ginsenosides in Saccharomyces cerevisiae was realized in this work and provides an alternative route compared to traditional extraction methods. Synthetic pathways of these three aglycons were constructed in S. cerevisiae by introducing β-amyrin synthase, oleanolic acid synthase, dammarenediol-II synthase, protopanaxadiol synthase, protopanaxatriol synthase and NADPH-cytochrome P450 reductase from different plants. In addition, a truncated 3-hydroxy-3-methylglutaryl-CoA reductase, squalene synthase and 2,3-oxidosqualene synthase genes were overexpressed to increase the precursor supply for improving aglycon production. Strain GY-1 was obtained, which produced 17.2 mg/L protopanaxadiol, 15.9 mg/L protopanaxatriol and 21.4 mg/L oleanolic acid. The yeast strains engineered in this work can serve as the basis for creating an alternative way for producing ginsenosides in place of extractions from plant sources.
A combined computational and experimental methodology is developed to predict new materials that should have desirable properties for CCS looping, and then select promising candidates to experimentally validate these predictions.
In recent years, an innovative public transportation (PT) mode known as the customized bus (CB) has been proposed and implemented in many cities in China to efficiently and effectively shift private car users to PT to alleviate traffic congestion and traffic-related environmental pollution. The route network design activity plays an important role in the CB operation planning process because it serves as the basis for other operation planning activities, for example, timetable development, vehicle scheduling, and crew scheduling. In this paper, according to the demand characteristics and operational purpose, a methodological framework that includes the elements of large-scale travel demand data processing and analysis, hierarchical clustering-based route origin-destination (OD) region division, route OD region pairing, and a route selection model is proposed for CB network design. Considering the operating cost and social benefits, a route selection model is proposed and a branch-and-bound-based solution method is developed. In addition, a computer-aided program is developed to analyze a real-world Beijing CB route network design problem. The results of the case study demonstrate that the current CB network of Beijing can be significantly improved, thus demonstrating the effectiveness of the proposed methodology.
Synergy is now a widely recognized approach that has direct applicability for new pharmaceuticals. The ethanolic extract of the aerial parts of the herb Sophora moorcroftiana showed significant antibacterial activity against drug-resistant Staphylococcus aureus, and its minimum inhibitory concentration (MIC) was 8 µg/mL. In a phytochemical study of the extract, five flavonoids were obtained. However, the isolates exhibited antibacterial activity in the range of 32-128 µg/mL, which was weaker than the extract. In combination with antibiotics, the antibacterially inactive compound genistein (1) and diosmetin (4) showed significant synergistic activity against drug-resistant S. aureus. In combination with norfloxacin, genistein (1) reduced the MIC to 16 µg/mL and showed synergy against strain SA1199B with a fractional inhibitory concentration index (FICI) of 0.38. With the antibiotics norfloxacin, streptomycin and ciprofloxacin, diosmetin (4) showed synergy against SA1199B, RN4220 and EMRSA-15, with FICI values of 0.38, 0.38 and 0.09, respectively. In an efflux experiment to elucidate a plausible mechanism for the observed synergy, genistein showed marginal inhibition of the NorA efflux protein.
This work not only leads to high yield production of rosmarinic acid and analogues, but also sheds new light on the construction of the pathway of rosmarinic acid in E. coli.
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