Integrated network pharmacology and metabolomics to reveal the mechanism of Pinellia ternata inhibiting non-small cell lung cancer cells.

Lung cancer is a malignant tumor with highly heterogeneous characteristics. A classic Chinese medicine, Pinellia ternata (PT), was shown to exert therapeutic effects on lung cancer cells. However, its chemical and pharmacological profiles are not yet understood. In the present study, we aimed to reveal the mechanism of PT in treating lung cancer cells through metabolomics and network pharmacology. Metabolomic analysis of two strains of lung cancer cells treated with Pinellia ternata extracts (PTE) was used to identify differentially abundant metabolites, and the metabolic pathways associated with the DEGs were identified by MetaboAnalyst. Then, network pharmacology was applied to identify potential targets against PTE-induced lung cancer cells. The integrated network of metabolomics and network pharmacology was constructed based on Cytoscape. PTE obviously inhibited the proliferation, migration and invasion of A549 and NCI-H460 cells. The results of the cellular metabolomics analysis showed that 30 metabolites were differentially expressed in the lung cancer cells of the experimental and control groups. Through pathway enrichment analysis, 5 metabolites were found to be involved in purine metabolism, riboflavin metabolism and the pentose phosphate pathway, including D-ribose 5-phosphate, xanthosine, 5-amino-4-imidazolecarboxyamide, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). Combined with network pharmacology, 11 bioactive compounds were found in PT, and networks of bioactive compound-target gene-metabolic enzyme-metabolite interactions were constructed. In conclusion, this study revealed the complicated mechanisms of PT against lung cancer. Our work provides a novel paradigm for identifying the potential mechanisms underlying the pharmacological effects of natural compounds.

The regulation of postharvest strawberry quality mediated by abscisic acid under elevated CO2 stress.

Elevated CO2 was a potential strategy for strawberry preservation. However, the regulatory mechanism remained unclear. In current study, transcriptome analysis showed that elevated CO2 played important roles in regulating strawberry fruit quality at the transcriptional level, and plant hormones metabolism at least partially involved in the regulatory process. Further, ABA was demonstrated to play important roles in the response to elevated CO2. Elevated CO2 inhibited the accumulation of ABA, which was 61% lower than that in control. Elevated CO2 repressed ABA synthesis by inhibiting NCED activity and the expression of FaNCED1/2, leading to the reduction of ABA accumulation as a result. Meanwhile, elevated CO2 also decreased ABA sensitivity by down-regulating FaSnRK2.4/2.6 and FaABI5 expression. The dual down-regulation of ABA signaling accounted for the regulation of fruit quality under elevated CO2 treatment. These results provide new insights into the mechanism of strawberry fruit response to elevated CO2.

Contribution of Cd passivating functional bacterium H27 to tobacco growth under Cd stress.

The cadmium (Cd) embedded in tobacco not only affects yield and quality but also harms human health. Microbial remediation has attracted widespread attention due to its low cost and minimal risk of secondary pollution. Therefore, researching microbes capable of inhibiting crop absorption of heavy metals or removing heavy metals from the environment has significant practical implications. This study screened a strain named H27 with a Cd immobilization efficiency of up to 76.60%. Static cultivation experiments showed that immobilization of Cd by H27 is achieved through intracellular absorption, hydroxyl, carboxyl, and phosphate group reactions on the cell wall. The bacterium can also secrete extracellular substances to adsorb Cd and increase the environmental pH, reducing the bioavailability of Cd. H27 reduced the accumulation of Cd in the stems of hydroponically grown tobacco by 55.23% and decreased the expression of three Cd transport genes, HAM2, IRT1, and NRAMP1, in the roots. Additionally, H27 increased the mineralization rate of organic matter, increased the content of humic acid in the soil, promoted the formation of smaller soil particles, and enhanced the adsorption and fixation of Cd by soil components while simultaneously raising the pH of rhizosphere and non-rhizosphere soils in tobacco growth environments. Both hydroponic and potted experiments showed that H27 alleviated the inhibitory effect of Cd on tobacco growth, significantly reducing Cd accumulation in various parts of tobacco and lowering the transfer coefficient of Cd within the tobacco plant. This study aims to effectively reduce the Cd content in tobacco using microbes, mitigate the harm of heavy metals in cigarettes to human health, and provide theoretical and practical basis for the application of microbial techniques to control heavy metal absorption in tobacco.

Transcriptomic analysis reveals the mechanism underlying the anthocyanin changes in Fragaria nilgerrensis Schlecht. and its interspecific hybrids.

BACKGROUND: Fragaria nilgerrensis (FN) provides a rich source of genetic variations for strawberry germplasm innovation. The color of strawberry fruits is a key factor affecting consumer preferences. However, the genetic basis of the fruit color formation in F. nilgerrensis and its interspecific hybrids has rarely been researched. RESULTS: In this study, the fruit transcriptomes and flavonoid contents of FN (white skin; control) and its interspecific hybrids BF1 and BF2 (pale red skin) were compared. A total of 31 flavonoids were identified. Notably, two pelargonidin derivatives (pelargonidin-3-O-glucoside and pelargonidin-3-O-rutinoside) were revealed as potential key pigments for the coloration of BF1 and BF2 fruits. Additionally, dihydroflavonol 4-reductase (DFR) (LOC101293459 and LOC101293749) and anthocyanidin 3-O-glucosyltransferase (BZ1) (LOC101300000), which are crucial structural genes in the anthocyanidin biosynthetic pathway, had significantly up-regulated expression levels in the two FN interspecific hybrids. Moreover, most of the genes encoding transcription factors (e.g., MYB, WRKY, TCP, bHLH, AP2, and WD40) related to anthocyanin accumulation were differentially expressed. We also identified two DFR genes (LOC101293749 and LOC101293459) that were significantly correlated with members in bHLH, MYB, WD40, AP2, and bZIP families. Two chalcone synthase (CHS) (LOC101298162 and LOC101298456) and a BZ1 gene (LOC101300000) were highly correlated with members in bHLH, WD40 and AP2 families. CONCLUSIONS: Pelargonidin-3-O-glucoside and pelargonidin-3-O-rutinoside may be the key pigments contributing to the formation of pale red fruit skin. DFR and BZ1 structural genes and some bHLH, MYB, WD40, AP2, and bZIP TF family members enhance the accumulation of two pelargonidin derivatives. This study provides important insights into the regulation of anthocyanidin biosynthesis in FN and its interspecific hybrids. The presented data may be relevant for improving strawberry fruit coloration via genetic engineering.

Forensic age estimation in adults by pubic bone mineral density using multidetector computed tomography.

Radiology plays a crucial role in forensic anthropology for age estimation. However, most studies rely on morphological methods. This study aims to investigate the feasibility of using pubic bone mineral density (BMD) as a new age estimation method in the Chinese population. 468 pubic bone CT scans from living individuals in a Chinese hospital aged 18 to 87 years old were used to measure pubic BMD. The BMD of the bilateral pubic bone was measured using the Mimics software on cross-sectional CT images and the mean BMD of the bilateral pubic bone was also calculated. Regression analysis was performed to assess the correlation between pubic BMD and chronological age and to develop mathematical models for age estimation. We evaluated the accuracy of the best regression model using an independent validation sample by calculating the mean absolute error (MAE). Among all established models, the cubic regression model had the highest R2 value in both genders, with R2 = 0.550 for males and R2 = 0.634 for females. The results of the best model test showed that the MAE for predicting age using pubic BMD was 8.66 years in males and 7.69 years in females. This study highlights the potential of pubic BMD as a useful objective indicator for adult age estimation and could be used as an alternative in forensic practice when other better indicators are lacking.

Crosstalk between melatonin and reactive oxygen species in fruits and vegetables post-harvest preservation: An update.

Fruits and vegetables contain numerous nutrients, such as vitamins, minerals, phenolic compounds, and dietary fibers. They reduce the incidence of cardiovascular diseases and the risk of certain chronic diseases, and improve the antioxidant and anti-inflammatory capacity. Moreover, melatonin was found in various fruits and vegetables species. Melatonin acts as a multifunctional compound to participate in various physiological processes. In recent years, many advances have been found that melatonin is also appraised as a key modulator on the fruits and vegetables post-harvest preservation. Fruits and vegetables post-harvest usually elicit reactive oxygen species (ROS) generation and accumulation. Excess ROS stimulate cell damage, protein structure destruction, and tissue aging, and thereby reducing their quality. Numerous studies find that exogenous application of melatonin modulates ROS homeostasis by regulating the antioxidant enzymes and non-enzymatic antioxidants systems. Further evidences reveal that melatonin often interacts with hormones and other signaling molecules, such as ROS, nitric oxide (NO), hydrogen sulfide (H2S), and etc. Among these 'new' molecules, crosstalks of melatonin and ROS, especially the H2O2 produced by RBOHs, are provided in fruits and vegetables post-harvest preservation in this review. It will provide reference for complicated integration of both melatonin and ROS as signal molecules in future study.

Dehydration effect of freeze-thaw on sludge: Temperature spatio-temporal distribution and multi-scale evaluation.

The freeze-thaw vacuum method for conditioning pretreated sludge has been proved that it not only has greater dewatering efficiency but also is more ecologically friendly. In this paper, the experiment is improved to address shortcomings in previous freeze-thaw vacuum approach for sludge treatment. The spatio-temporal distribution relationship of distance-time-temperature is developed and divided into two stages by numerically fitting the temperature change of freezing tubes in the sludge. It is expected to guide the time control of large-scale frozen sludge in practical engineering applications to achieve optimal dewatering treatment. Furthermore, the performance of dehydration after the model test is evaluated on multi-scale: settlement and mechanical properties (macroscopic perspective), mean particle size (mesoscopic perspective), and SEM microstructure (microscopic perspective). The results reveal that the improved sludge treatment method of alternating freeze-thaw vacuum procedures, using both prefabricated horizontal drains (PHDs) and prefabricated vertical drains (PVDs), substantially benefits the sludge dewatering and reduction. This method results in an unparalleled volume reduction of 63.51% and a water content reduction to 58.54%. Moreover, in-situ vane shearing strength of the sludge obtained from the improved test meets the strength requirement for the landfill final cover soil, demonstrating that the method is superior in improving mechanical properties.

Downregulation of C12orf75 gene inhibits migration and invasion of liver cancer cell via suppressing the Wnt/ß-catenin signaling pathway in vitro.

Hepatic carcinoma (HCC) is the second leading cause of death in cancer patients, and its incidence is on the rise. The prerequisite for the deterioration of liver cancer is the malignant migration and invasion of cancer cells, and in this study the C12orf75 gene was firstly showed that it has high expression characteristics in six HCC cell lines. By bioinformatics analysis, the up-regulation of C12orf75 was negatively correlated with the survival rate of HCC patients. CCK-8 and flow cytometry experiments indicated C12orf75 had no effect on cell proliferation, apoptosis and cycle of HuH-7 and Hep G2 cell lines, respectively. However, transwell assay showed that downregulation of C12orf75 gene could obviously inhibit invasion and migration of liver cancer cell. At the same time, this regulation process has been verified to involve the participation of Wnt/ß-catenin signaling pathway. As a result, all these results showed that C12orf75 gene was a oncogene in liver cancer, which maybe a novel screening and diagnosing biomarker for patients with liver cancer, and this is of great significance for the development of targeted drugs for liver cancer patients.

Astragalus Polysaccharide Suppresses Cell Proliferation and Invasion by Up-Regulation of miR-195-5p in Non-small Cell Lung Cancer.

Non-small cell lung cancer (NSCLC) accounts for approximately 85% of lung cancer-related mortality, and it has a high risk of early recurrence and distant metastasis. The prerequisite for the deterioration of NSCLC is the malignant proliferation and migration of cancer cells, and in this study Astragalus membranaceus polysaccharides (APS) was firstly showed that it could decrease the cell proliferation of A549 and NCI-H1299. Through bioinformatics analysis, the up-regulation of miR-195-5p was positively correlated with the survival rate of lung cancer patients. Real-time PCR indicated APS could increase the expression level of miR-195-5p, and the miR-195-5p inhibitor was used to verify that it could reverse the inhibitory effect of Astragalus polysaccharide on lung cancer cell migration and invasion. Therefore, we believe that APS could inhibit the proliferation and migration of NSCLC cells by regulating miR-195-5p, which laid the foundation for further research on the functional mechanism of miR-195-5p in NSCLC.

Machine-learning assisted modelling of multiple elements for authenticating edible animal blood food.

Elemental fingerprint coupled with machine learning modelling was proposed for species authentication of the edible animal blood gel (EABG). A total of 25 elements were determined by inductively coupled plasma mass spectrometry (ICP-MS) and atomic absorption spectroscopy (AAS) in 150 EABG samples prepared from five species of animals, namely duck, chicken, bovine, pig, and sheep. Extreme learning machine (ELM) models were constructed and optimized. Principal component analysis and Fisher linear discriminant analysis were comparatively utilized for dimension reduction of the crucial input elements selected via stepwise discriminant analysis and one-way ANOVA. The optimal ELM model was obtained with the crucial elements selected by one-way ANOVA from the relative content of the measured elements, which afforded accuracies of 98.0% and 96.0% for the training and test set, respectively. All findings suggest that elemental fingerprint accompanied by ELM have great potential in authenticating the edible animal blood food.

Forensic age prediction and age classification for critical age thresholds via 3.0T magnetic resonance imaging of the knee in the Chinese Han population.

OBJECTIVES: To assess the performance of knee MRI for forensic age prediction and classification for 12-, 14-, 16-, and 18-year thresholds. METHODS: The ossification stages of distal femoral epiphyses and proximal tibial epiphyses were assessed using an integrated staging system by Schmeling et al. and Kellinghaus et al. for knee 3.0T MRI with T1-weighted turbo spin-echo (T1-TSE) in sagittal orientation among 852 Chinese Han individuals (483 males and 369 females) aged 7-30 years. Regression models for age prediction were constructed and their performances were evaluated based on mean absolute deviation (MAD) values. In addition, the performances of age classification were assessed using receiver operating characteristic (ROC) analyses. RESULTS: The intra- and inter-observer agreement levels were very good (κ > 0.80). The complete fusion of those two types of epiphyses took place before 18.0 years in our study participants. The minimum MAD values were 2.51 years (distal femur) and 2.69 years (proximal tibia) in males, and 2.75 years (distal femur) and 2.87 years (proximal tibia) in females. The specificity values of constructed prediction models were all above 90% for the 12-, 14-, and 16-year thresholds, compared to the 74.8-84.6% for the 18-year threshold. Better performances of age prediction and classification were observed in males by distal femoral epiphyses. CONCLUSIONS: Ossification stages via 3.0T MRI of the knee with T1-TSE sequence using an integrated staging system could be a reliable noninvasive method for age prediction or for age classification for 12-, 14-, and 16-year thresholds, especially in males by distal femoral epiphyses. However, assessments based on the full bony fusion of the distal femoral epiphysis and proximal tibial epiphysis seemed not reliable for age classification for the 18-year threshold in the Chinese Han population.

Insights into vacuum preloading consolidation of landfill sludge based on Fe2+-activated sodium persulfate.

To further reduce the in-situ sludge from landfill, Fe2+-activated sodium persulfate combined with vacuum preloading was first proposed. Firstly, the effects of optimal Na2S2O8 dosage on the landfill sludge (LS) were investigated by the vacuum filtration experiments. Then, vacuum preloading experiments were conducted on the sludge with different Na2S2O8 dosages to study the water content, water discharge, and settlement. Besides, sludge particle size diversification was carried out by particle size distribution and scanning electron microscopy (SEM) tests. The results were summarized as follows: the specific resistance of filtration (SRF) of LS could be reduced by 98.5% mostly when the Na2S2O8 dosage was 30%; the particle size became significantly smaller, and large particles were converted to small particles; the water content dropped from 86.9 to 58.3%; and the SEM test manifested the oxidation of sodium persulfate caused the destruction of the glial structure of the sludge and the recombination of partial particles.

Freeze-thaw vacuum treatment of landfill sludge: Mechanism of uneven frost heaving and dewatering performance.

The method of freeze-thaw combined with vacuum pretreatment for landfill sludge (LS) has attracted extensive attention. However, most of the existing approaches are based on small-scale laboratory testing, and further testing studies must be performed to realize in situ treatment. To enhance the practicality of such approaches, the range of temperature effects on LS was analysed after field freeze-thaw model testing. After the freeze-thaw model test, samples were transported to the laboratory for unidirectional oedometer tests, and the remaining samples were retained in the field to continue vacuum model testing for exploring the differences in the consolidation and drainage effect of the LS. Results show that temperature changes during freeze-thaw process affect the distribution of sludge and water in the model boxes, resulting in frost heave and the appearance of "extrusion rings". In addition, the coefficient of consolidation obtained from the unidirectional oedometer test shows that the consolidation coefficient is generally larger near the freezing tubes at a lower temperature. The settlement determined from the field vacuum preloading test shows that the subsequent vacuum consolidation settlement is larger at the position with a lower elevation of the frozen sludge surface. The comparison indicates that the consolidation and drainage effect in the field is not as significant as that in the laboratory. The findings can provide reference to optimize the field conditions during the in situ engineering practice of sludge treatment.

Quantitative Assessment of Abiotic Stress on the Main Functional Phytochemicals and Antioxidant Capacity of Wheatgrass at Different Seedling Age.

The wheat seedlings of 6 days old were daily subjected to ultraviolet irradiation (irradiating for 5, 10, 20, 40, and 60 min/day, respectively), Polyethylene glycol 6000 (5, 10, 15, 20, 25% in 1/2 Hoagland solution, respectively), and salinity solution (10, 25, 50, 100, 200 mM in 1/2 Hoagland solution, respectively), while the control group (CK) was supplied only with the Hoagland solution. The wheatgrass was harvested regularly seven times and the total soluble polysaccharides, ascorbic acid, chlorophyll, total polyphenol, total triterpene, total flavonoid, and proanthocyanins content were tested. The antioxidant capacity was evaluated through 2,2'-azino-bis (3-ethylbenzthia-zoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging ability, and ferric ion reducing power. Technique for order preference by similarity to ideal solution (TOPSIS) mathematical model was adopted to comprehensively assess the functional phytochemicals of the different treatments. The results showed that the accumulation patterns of phytochemicals under abiotic stress were complex and not always upregulated or downregulated. The antioxidant activity and functional phytochemicals content of wheatgrass were significantly affected by both the stress treatments and seedling age, while the latter affected the chemicals more efficiently. The top five highest functional phytochemicals were observed in the 200 mM NaCl treated group on the 21st and 27th day, 25% PEG treated group on the 24th day, 200 mM NaCl treated group on the 24th day, and the group of 40 min/day ultraviolet exposure on 27th day.

Novel insights into enhanced dewaterability and consolidation characteristics of landfill sludge and fresh sludge conditioned by Fe2+ activated sodium persulfate.

Considering the reduction and resource utilization of landfill sludge (LS) and fresh sludge (FS), Fe2+ activated Na2S2O8 is proposed. The effects of the molar ratio of Fe2+/S2O82- and the addition of Na2S2O8 on the dewatering performance of sludge were studied by vacuum filtration experiments. Consolidation tests were conducted on the sludge with different Na2S2O8 dosage, and the compression, consolidation, and permeability characteristics of the sludge were researched. Besides, via particle size distribution (PSD) and scanning electron microscope (SEM) test, the variation of particle size of sludge was studied from the microscopic perspective. The results are as follows: the specific resistance of filtration (SRF) of LS and FS decreases by 99.3%, 95.2% at an optimal dosage (the molar ratio of (Fe2+/S2O82-) = 1, 30% Na2S2O8); the particle size of LS and FS is significantly smaller; the consolidation and permeability coefficients are increased by 1-2 orders of magnitude compared with non-conditioned sludge; the water content of LS and FS drops from 86.5% to 58.4%, 82.4%-59.7%. The research results have certain guiding significance for the in-situ treatment of sludge deep dewatering.

Oxidative stress and mitochondrial membrane potential are involved in the cytotoxicity of perfluorododecanoic acid to neurons.

Perfluorododecanoic acid (PFDoA), used in numerous commercial products, was recently demonstrated to accumulate in the brain more easily than other perfluorinated compounds and to cause cognitive deficits. In this study, pheochromocytoma 12 (PC12) cells were exposed to doses of PFDoA to explore the cytotoxicity of this compound to neurons. The results showed that treatment with PFDoA decreased PC12 cell viability dose-dependently. Treatment with 50 and 100 µM PFDoA significantly increased reactive oxygen species (p < 0.01) and malondialdehyde (p < 0.01) and decreased total antioxidant capacity (p < 0.05 and p < 0.01, respectively) in PC12 cells. The administration of 50 and 100 µM PFDoA led to a loss of mitochondrial membrane potential (MMP) (p < 0.05 and p < 0.01, respectively) in PC12 cells. The activity of caspase 3 was obviously increased (p < 0.05) in 100 µM PFDoA-treated PC12 cells. In general, the results demonstrated that PFDoA exposure could result in the disruption of MMP, which may contribute to the increase of oxidative stress and activation of the apoptotic signaling cascade in PC12 cells.

Ultrasound enhanced biosynthesis of L-theanine from L-glutamine and ethylamine by recombinant γ-glutamyltranspeptidase.

A mutant library of the key amino acid residue site E387 in γ-glutamyltranspeptidase was constructed to screen the mutant enzymes with significantly improved thermal stability (E387Q). The reaction temperature of the mutant enzyme (E387Q) was 10℃ higher than that of the parent enzyme. Ultrasound-assisted synthesis of L-theanine by γ-glutamyltranspeptidase was investigated. The effects of ultrasonic power, reaction pH and substrate concentration on the enzymatic synthesis of L-theanine were studied by the response surface method. The results showed that the optimal process conditions are ultrasonic power of 100 W, reaction pH of 9, substrate L-glutamine concentration of 120 mmol/L, reaction temperature of 45℃, and L-theanine yield of 89.1%. The yield of L-theanine is 2.61 times higher than that obtained without ultrasound. Ultrasound can significantly promote the synthesis of L-theanine by γ-glutamyltranspeptidase.

[Metagenomic Analysis of Antibiotic Resistant Bacteria and Resistance Genes in a Pharmaceutical and Chemical Wastewater Treatment Plant].

Wastewater treatment plants hold a vast pool of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). The aim of this study is to analyze the ARB and ARGs in a pharmaceutical and chemical wastewater treatment plant using a metagenomic technique. The results of taxonomic annotation revealed that bacteria were the predominant domain. The most abundant phyla and genus was Proteobacteria and Hyphomicrobium, respectively. A total of 74 categories of ARGs were predicted using CARD with the most dominant types being sav 1866, dfrE, and mfd. Furthermore, a network analysis was conducted to investigate the co-occurrence patterns between ARGs and microbial taxa. ARGs were found to be highly connected to microbial taxa at the genus level. With respect to the antibiotic resistance mechanisms, antibiotic-specific efflux pumps appeared to be the most common mechanisms. Among these, resistance-nodulation-cell division (RND) was the major type. The most important functional pathway of this microbial community was metabolic correlation. Interestingly, there were many genes related to human diseases, among which bacterial infectious diseases were the main ones. On the one hand, these data further confirmed that pharmaceutical and chemical wastewater treatment plants are rich in ARB and ARGs. The accumulation of ARGs increases the potential environmental risks, and hence it is necessary to strengthen the active monitoring of ARB and ARGs in pharmaceutical and chemical wastewater treatment plants. On the other hand, research on ARB and ARGs offers important information for the selection of deep processing technology to effectively remove ARB and ARGs.

Downregulation of GATS gene inhibits proliferation, clonogenicity and migration in triple negative breast cancer cells MDA-MB-231 by cell autophagy.

The triple negative breast cancer (TNBC) accounts for 15% to 20% of the total number of breast cancer diagnosed. A number of clinical studies have shown that TNBC has a high risk of early recurrence and distant metastasis, and a low rate of disease free survival and total survival. The premise of TNBC deterioration was abnormal proliferation and migration of tumor cells, and this study firstly showed that GATS gene could promote proliferation of MDA-MB-231 breast cancer cells. Through lentiviral expression system, the GATS gene was konckdown by shGATS lentivirus infection in the MDA-MB-231 cells, and the result indicated it could remarkably decrease the ability of cell proliferation and migration. Real-time PCR, western blot and immunofluorescence experiments showed the expressions of protein LC3, and p-Akt in shGATS cell group were lower than the shCtrl group. Therefore, we suggest the GATS could promote the MDA-MB-231 cell proliferation, migration and clonogenicity through cell autophagy by the PI3K/Akt pathway, which paved the way for further study the function of GATS in TNBC, and GATS may potentially be a target for gene therapy against triple negative breast cancer.

Highly efficient preparation of active S-phenyl-L-cysteine with tryptophan synthase using a chemoenzymatic method.

BACKGROUND: S-Phenyl-L-cysteine is regarded as having potential applicability as an antiretroviral/protease inhibitor for human immunodeficiency virus (HIV). In the present study, optically active S-phenyl-L-cysteine was prepared in a highly efficient manner from inexpensive bromobenzene using tryptophan synthase through a chemoenzymatic method. RESULTS: The chemoenzymatic method used a four-step reaction sequence. The process started with the reaction of magnesium and bromobenzene, followed by a Grignard reaction, and then hydrolysis and enzymatic synthesis using tryptophan synthase. Through this approach, S-phenyl-L-cysteine was chemoenzymatically synthesized using tryptophan synthase from thiophenol and L-serine as the starting material. CONCLUSIONS: High-purity, optically active S-phenyl-L-cysteine was efficiently and inexpensively obtained in a total yield of 81.3% (> 99.9% purity).