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Both natural ginsenoside F2 and unnatural ginsenoside 3β,20S-Di-O-Glc-DM were reported to exhibit anti-tumor activity. Traditional approaches for producing them rely on direct extraction from Panax ginseng, enzymatic catalysis or chemical synthesis, all of which result in low yield and high cost. Metabolic engineering of microbes has been recognized as a green and sustainable biotechnology to produce natural and unnatural products. Hence we engineered the complete biosynthetic pathways of F2 and 3β,20S-Di-O-Glc-DM in Saccharomyces cerevisiae via the CRISPR/Cas9 system. The titers of F2 and 3β,20S-Di-O-Glc-DM were increased from 1.2 to 21.0 mg/L and from 82.0 to 346.1 mg/L at shake flask level, respectively, by multistep metabolic engineering strategies. Additionally, pharmacological evaluation showed that both F2 and 3β,20S-Di-O-Glc-DM exhibited anti-pancreatic cancer activity and the activity of 3β,20S-Di-O-Glc-DM was even better. Furthermore, the titer of 3β,20S-Di-O-Glc-DM reached 2.6 g/L by fed-batch fermentation in a 3 L bioreactor. To our knowledge, this is the first report on demonstrating the anti-pancreatic cancer activity of F2 and 3β,20S-Di-O-Glc-DM, and achieving their de novo biosynthesis by the engineered yeasts. Our work presents an alternative approach to produce F2 and 3β,20S-Di-O-Glc-DM from renewable biomass, which lays a foundation for drug research and development.
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Medicinal natural products derived from plants are usually of low content and difficult to extract and isolate. Moreover, these compounds are structurally complex, making it difficult to obtain them by environmental unfriendly chemical synthesis. Biosynthesis of medicinal natural products through synthetic biology is a novel, environment-friendly and sustainable approach. Taking terpenoids (ginsenosides, paclitaxel, artemisinin, tanshinones), alkaloids (vincristine and morphine), and flavonoids (breviscapine) as examples, this review summarizes the advances of the biosynthetic pathways and synthetic biology strategies of plant-derived medicinal natural products. Moreover, we introduce the key technologies and methods of synthetic biology used in the research of medicinal natural products, and provide future prospects in this area.
Subject(s)
Biological Products , Biosynthetic Pathways , Plants , Synthetic Biology , TerpenesABSTRACT
OBJECTIVE@#To observe the changes of blood pressure and S-100B, neuron specific enolase (NSE) protein in hypertensive dogs with high fat diet after catheter-based renal sympathetic denervation.@*METHODS@#Twelve Beagles were divided into an interventional group (n=6) and a sham-operation group (n=6). After baseline measurements, the Beagles were fed with lard oil for 3 months. After 3 months, the interventional group had renal sympathetic denervation by percutaneous catheter based radiofrequency ablation and the control group had renal angiography. The blood pressure, plasma S-100B, and NSE before the operation and 3 days, 1 week, 2 weeks, 1 month, 2 months and 3 months after the operation were measured.@*RESULTS@#The dogs had significantly higher levels of systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MBP) compared to its baselines (P<0.05). The SBP, DBP and MBP in the interventional group were significantly lower than those in the control group 1 month and 3 months after the operation (P<0.05). Three months after the operation, renal angiography in all dogs revealed no sign of renal artery stenosis. Plasma S-100B and NSE expression in the interventional group were higher than those in the control group 3 days, 1 week and 2 weeks after the operation (P<0.05).@*CONCLUSION@#Renal sympathetic denervation can significantly reduce the SBP, DBP and MBP in hypertensive dogs. The plasma S-100B and NSE may be used as indicators for assessment of renal nerve injury after renal sympathetic denervation.
Subject(s)
Animals , Dogs , Blood Pressure , Catheter Ablation , Hypertension , Metabolism , Kidney , General Surgery , Phosphopyruvate Hydratase , Metabolism , S100 Calcium Binding Protein beta Subunit , Metabolism , SympathectomyABSTRACT
Objective To investigate the predictive value of cardiovascular risk factors in coronary atherosclerotic heart disease.Methods A total of 400 patients who underwent coronary angiography (CAG) in our hospital was divided into multiple vascular lesion group,single vascular lesions group,and non-CAD group according to the result of CAG and their clinical data were analyzed retrospectively.The correlation between cardiovascular risk factors and coronary artery lesions was analyzed,and the independent risk factor of CAD was screened by multi-factorial logistic regression analysis.Results There was significant difference in total cholesterol (TC),triglycerides (TG),high density lipoprotein cholesterol (HDLC),fasting blood glucose (FBG),inflammatory cells,carotid artery plaque,brachial-ankle pulse wave velocity (baPWV),high-sensitivity C-reactive protein (Hs-CRP) between the CAD group and the non-CAD group (P ＜ 0.05 or P ＜ 0.01).The number of coronary artery lesion branch was increased significantly when risk factors,such as age,body mass index(BMI),hypertension,diabetes mellitas,smoking,carotid artery plaque,TG,TC,low density lipoprotein cholesterol (LDL-C),FBG,WBC,monocytes (M),neutrophils (N),neutrophils/lymphocytes (N/L),baPWV,and Hs-CRP.Other risk factors including TC,HDL-C,L were decreased with a statistically significant difference (P ＜ 0.05).There was no significant relation among,and left ventricular ejection fraction (LVEF)％.The most significant risk factor was carotid artery plaque that was independently associated with coronary heart disease (b =1.264,P ＜ 0.01),followed by smoking (b =1.204,P ＜0.01),HDL-C (b =1.104,P ＜0.01),TC (b =1.082,P ＜0.01) diabetes mellitus (b =0.956,P ＜0.01),baPWV increased (b =0.741,P ＜0.01),WBC (b=0.721,P ＜0.01),hypertension (b =0.602,P ＜0.01),the age (b =0.538,P ＜0.01),and HsCRP(b =0.421,P ＜ 0.01).Conclusions The results suggest that the hypertension,hyperlipidemia,smoking,age,baPWV,inflammatory cells,Hs-CRP,and carotid artery plaque was a significant independent CHD risk factors.
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Aim To investigate the effect of tanshinoneⅡA(TSN) on the hypertrophy induced by angiotensinⅡ(AngⅡ)in the primary culture of neonatal rat cardiomyocytes. Methods Cardiomyocytes of Wistar rats were cultured with pancreatin and preplating technique. The hypertrophy of cardiomyocytes was induced with angiotensinⅡ, and intervened with TanshinoneⅡA and Valsartan. The effect of TSN on cardiomyocytewas evaluated by 3-[4,5-dimethylthiazol-2-yl]-3,5-diphenylformazan (MTT) assay. As the index of cardiomyocyte hypertrophy, protein synthesis rate was measured by ~3H-Leucine incorporation and the cell size was determined by phase contrast microscope. The proto-oncogene c-fos、c-myc and c-jun mRNA expressions were assessed using reverse transcription polymerase chain reaction (RT-PCR). Results Exposure of cultured cardiomyocytes to TSN (5～80 ?mol?L~ -1 ) for 24 h produced marginal cytotoxicity. Additonally, myocyte monolayers continued to contract synchronously in the presence of TSN. With the treatment of cardiomyocytes by AngII for 7 days, the cell size of cardiomyocytes of the group of Ang Ⅱ(28.5?3.8) ?m increased more prominently than the group of control(19.8?1.9) ?m(P
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Aim To observe effects of Sodium tanshinoneⅡA sulfonate(STS) on angiotensionⅡ(AngⅡ)-induced cardiomyocyte hypertrophy and the expression of phosphorylated extracellular signal-regulated kinase1/2 (p-ERK1/2). Methods In the primary culture of neonatal rat cardiomyocytes, as indexes of cardiomyocyte hypertrophy, the total protein was determined by coomassie brilliant blue and protein synthesis rate was measured by [3H]-Leucine incorporation. The expression of p-ERK1/2 was assessed using Western blot and fluorescence microscope. Results ① The total protein and protein synthesis rate stimulated by Ang Ⅱ(1 ?mol?L-1)in the cardiomyocytes increased significantly in contrast to that of control; STS could effectively decrease the increased total protein level induced by Ang Ⅱand markedly inhibit synthesis of protein. ② AngⅡ(1 ?mol?L-1) had the effect of promoting activation of ERK1/2 and then appeared in nucleus rapidly. The translocation process of ERK1/2 induced by AngⅡ was blocked distinctly by STS. ③ Cardiomyocyte pretreated with Ang Ⅱ(1 ?mol?L-1)for 5 min, the p-ERK1/2 protein expression began to increase ,the peak effect was at 10 min. While pretreatment with STS(2, 10, 50 ?mol?L-1) ,Ang Ⅱ-induced increase in p-ERK1/2 were inhibited evidently. ④ In pretreatment of cardiomyocyte with STS in different doses for 30 min, STS was found to be able to inhibit the expression of p-ERK1/2 stimulated by AngⅡ in a dose-dependent manner. Conclusions The results suggested that activation of ERK1/2 might play an important role in cardiomyocytes hypertrophy induced by Ang Ⅱ,and the anti-hypertrophic effect of STS on cardiomyocyte hypertrophy induced by AngⅡ might be associated to its inhibitory effect on ERK signaling pathway.