Sustainable production of natural products using synthetic biology: Ginsenosides.

Synthetic biology approaches offer potential for large-scale and sustainable production of natural products with bioactive potency, including ginsenosides, providing a means to produce novel compounds with enhanced therapeutic properties. Ginseng, known for its non-toxic and potent qualities in traditional medicine, has been used for various medical needs. Ginseng has shown promise for its antioxidant and neuroprotective properties, and it has been used as a potential agent to boost immunity against various infections when used together with other drugs and vaccines. Given the increasing demand for ginsenosides and the challenges associated with traditional extraction methods, synthetic biology holds promise in the development of therapeutics. In this review, we discuss recent developments in microorganism producer engineering and ginsenoside production in microorganisms using synthetic biology approaches.

Discrimination of Lycium chinense and L. barbarum Based on Metabolite Analysis and Hepatoprotective Activity.

Lycii Fructus is a traditional medicine used to prevent liver and kidney diseases, which commonly derives from Lycium chinense and Lycium barbarum. Here, the extracts and ethyl acetate-soluble fractions of L. chinense fruits exhibited better hepatoprotective effects than those of L. barbarum, which was likely due to differences in their composition. Therefore, GC-MS and HPLC analyses were conducted to characterize the metabolite differences between L. chinense and L. barbarum. Based on amino acid (AA) and phenolic acid (PA) profiling, 24 AAs and 9 PAs were identified in the two species. Moreover, each species exhibited unique and readily distinguishable AA and PA star graphic patterns. HPLC analysis elucidated composition differences between the ethyl acetate-soluble layers of the two compounds. Further, NMR analysis identified their chemical structures as 4-(2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl)butanoic acid and p-coumaric acid. The higher content of 4-(2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl)butanoic acid was detected in L. chinense, whereas the content of p-coumaric acid was higher in L. barbarum. Therefore, the differences in the relative contents of these two secondary metabolites in the ethyl acetate-soluble layer of Lycii Fructus could be a good marker to discriminate between L. chinense and L. barbarum.

Fermented Korean Red Ginseng Extract Enriched in Rd and Rg3 Protects against Non-Alcoholic Fatty Liver Disease through Regulation of mTORC1.

The fermentation of Korean red ginseng (RG) increases the bioavailability and efficacy of RG, which has a protective role in various diseases. However, the ginsenoside-specific molecular mechanism of the fermented RG with Cordyceps militaris (CRG) has not been elucidated in non-alcoholic fatty liver disease (NAFLD). A mouse model of NAFLD was induced by a fast-food diet (FFD) and treated with CRG (100 or 300 mg/kg) for the last 8 weeks. CRG-mediated signaling was assessed in the liver cells isolated from mice. CRG administration significantly reduced the FFD-induced steatosis, liver injury, and inflammation, indicating that CRG confers protective effects against NAFLD. Of note, an extract of CRG contains a significantly increased amount of ginsenosides (Rd and Rg3) after bioconversion compared with that of conventional RG. Moreover, in vitro treatment with Rd or Rg3 produced anti-steatotic effects in primary hepatocytes. Mechanistically, CRG protected palmitate-induced activation of mTORC1 and subsequent inhibition of mitophagy and PPARα signaling. Similar to that noted in hepatocytes, CRG exerted anti-inflammatory activity through mTORC1 inhibition-mediated M2 polarization. In conclusion, CRG inhibits lipid-mediated pathologic activation of mTORC1 in hepatocytes and macrophages, which in turn prevents NAFLD development. Thus, the administration of CRG may be an alternative for the prevention of NAFLD.

Reduced sludge production in a membrane bioreactor by uncoupling metabolism and its effect on phosphorus accumulation in the biomass.

Due to its limited recycling or reuse, treatment and disposal of excess waste activated sludge has been a major challenge. As a preemptive method, therefore, uncoupling metabolism for reduced sludge production has been investigated recently. In this study, we operated a pilot-scale A2O-membrane bioreactor (MBR) system incorporating an anaerobic sludge holding tank (SHT) in a sludge recycling line to induce uncoupling metabolism, and investigated sludge production and treatment efficiency. After operation for ≥1 year, the Yobs value was estimated to be 0.041 g mixed liquor suspended solid (MLSS)/g chemical oxygen demand with 198.7 days of solids retention time (SRT). This Yobs value was markedly lower than those reported previously. Since MBR can be operated with a relatively high MLSS and prolonged SRT, the greatest reduction was achieved by combination with uncoupling metabolism. Phosphate fractionation experiments of the MLSS from the pilot MBR suggested the total phosphate content of microorganisms was 47.0 mg P/g mixed liquor volatile suspended solid; 83% higher than that of the activated sludge process and 49% higher than that of the conventional A2O process. Of the increased phosphate contents, that of the acid-insoluble polyphosphate (AISP) fraction was greatest, suggesting that growth inhibition by uncoupling metabolism stimulates AISP synthesis, which can be utilized under growth-limiting conditions. ABBREVIATIONS: AISP: acid-insoluble polyphosphate; ASP: acid-soluble polyphosphate; BNR: biological nutrients removal; EPS: extracellular polymeric substance; LMH: L/m2 h; MBR: membrane bioreactor; OST: oxic-settling-anaerobic; PAO: phosphate accumulation organism; PCA: perchloric acid; SBR: sequencing batch reactor; SHT: sludge holding tank; SRT: solids retention time; TN: total nitrogen; TP: total phosphate; WAS: waste activated sludge.

Spectrophotometric measurement of minimal erythema dose sites after narrowband ultraviolet B phototesting: clinical implication of spetrophotometric values in phototherapy.

BACKGROUND: The spectrophotometer is well known to be a useful tool for estimating the objective minimal erythema dose (MED) during planning of phototherapy protocol. However, only a few spectrophotometric values are used to evaluate the erythema and pigmentation of the MED site during phototesting. OBJECTIVE: To determinea new meaning of the relationships among spectrophotometric values during phototesting. METHODS: Twenty-five patients with psoriasis and 23 patients with vitiligo were selected before undergoing narrowband ultraviolet B phototherapy. We interpreted the gross findings of erythema and measured the L(*)a(*)b(*) values using a spectrophotometer at each phototest spot. We compared MEDs, basic spectrophotometric values (L(*)a(*)b(*)), and b(*)/L(*) values separately according to skin type, and determined the correlation of each spectrophotometric value and the correlation between a(*) and b(*)/L(*) values. RESULTS: Among L(*)a(*)b(*) values, only b(*) values showed a statistically significant difference between the type III and IV groups (p=0.003). There was a positive correlation only between MEDs and b(*) values (p<0.05). The average b(*)/L(*)value in the type IV group was significantly higher than the type III group (p<0.05). CONCLUSION: The higher b(*) values in type IV skin indicates that skin tanning develops more prominently than type III. The correlation between MEDs and b(*) values may signify that the skin pigmentation status is deepened with the higher MEDs. The difference in b(*)/L(*)values between type III and IV skin reflects that the b(*)/L(*)value is thought to be an index of tanning. The a(*) value, known as an index of erythema, does not influence the degree of tanning.

Water quality of runoff from agricultural-forestry watersheds in the Geum River Basin, Korea.

Forestry and agricultural land uses constitute 85% of Korea and these land uses are typically mixed in many watersheds. Land cover is one of the most important factors affecting diffuse pollution and water quality. The aim of this study is to estimate the pollutant concentrations in runoff from four study watersheds consisting of a mix of forestry and agricultural land uses at different ratios in the Geum River Basin. The effect of topographical variables was also considered. The ratio of agricultural land use to the total area of study watersheds was in the range of 0.01-0.36. Flow rate and water quality (suspended solids, organics and nutrients) of runoff from 40 rainfall events were monitored at the study watersheds. Descriptive statistics showed higher nutrients and organic concentrations in runoff from watershed with higher agricultural activities. Event Mean Concentration (EMC) of individual runoff event was calculated for each water quality constituent based on the flow rate and concentration data of runoff discharge, and arranged on a cumulative probability scale according to runoff occurrence. From the correlation analysis between EMC data and affecting variables, the ratio of agricultural land use to the total area was identified as the parameter that most affected the magnitude of EMC.