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Science Advances Aug 2023Developing technologies based on the concept of methanol electrochemical refinery (e-refinery) is promising for carbon-neutral chemical manufacturing. However, a lack of...
Developing technologies based on the concept of methanol electrochemical refinery (e-refinery) is promising for carbon-neutral chemical manufacturing. However, a lack of mechanism understanding and material properties that control the methanol e-refinery catalytic performances hinders the discovery of efficient catalysts. Here, using O isotope-labeled catalysts, we find that the oxygen atoms in formate generated during the methanol e-refinery reaction can originate from the catalysts' lattice oxygen and the O-2p-band center levels can serve as an effective descriptor to predict the catalytic performance of the catalysts, namely, the formate production rates and Faradaic efficiencies. Moreover, the identified descriptor is consolidated by additional catalysts and theoretical mechanisms from density functional theory. This work provides direct experimental evidence of lattice oxygen participation and offers an efficient design principle for the methanol e-refinery reaction to formate, which may open up new research directions in understanding and designing electrified conversions of small molecules.
PubMed: 37624888
DOI: 10.1126/sciadv.adh9487 -
Internal Medicine (Tokyo, Japan) Apr 2024
PubMed: 38569902
DOI: 10.2169/internalmedicine.3450-24 -
Biotechnology For Biofuels and... Nov 2023Single-cell protein (SCP) production in the methylotrophic yeast Pichia pastoris has the potential to achieve a sustainable protein supply. However, improving the...
Single-cell protein (SCP) production in the methylotrophic yeast Pichia pastoris has the potential to achieve a sustainable protein supply. However, improving the methanol fermentation efficiency and reducing carbon loss has been a long-standing challenge with far-reaching scientific and practical implications. Here, comparative transcriptomics revealed that PAS_0305, a gene directly associated with cell wall thickness under methanol stress, can be used as a target for unlocking cell wall sensors. Intracellular trehalose accumulation confirmed that cell wall sensors were activated after knocking out PAS_0305, which resulted in increased cell wall permeability. Genome-wide signal perturbations were transduced through the HOG module and the CWI pathway, which was confirmed to connected by Pbs2-Mkk. As a consequence of CWI pathway activation, ΔPAS_0305 elicited a rescue response of cell wall remodeling by increasing the β-1,3-glucan content and decreasing the chitin/mannose content. Remarkably, perturbations in global stress signals led to a fine-tuning of the metabolic network of ΔPAS_0305, resulting in a superior phenotype with highest crude protein and methanol conversion rate of 67.21% and 0.46 gDCW/g. Further genome-scale metabolic models were constructed to validate the experimental results, confirming that unlocking cell wall sensors resulted in maximized flux from methanol towards SCP and effectively addressing the issue of carbon loss in methanol fermentation. This work sheds new light on the potential of manipulating cellular signaling pathways to optimize metabolic networks and achieve exceptional phenotypic characteristics, providing new strategies for constructing versatile cell factories in P. pastoris.
PubMed: 37978550
DOI: 10.1186/s13068-023-02428-7 -
Journal of Tropical Medicine 2023The emergence of multidrug bacterial resistance poses a great public health problem and requires a constant search for new antibacterial agents. However, Niger's flora...
The emergence of multidrug bacterial resistance poses a great public health problem and requires a constant search for new antibacterial agents. However, Niger's flora possesses several medicinal plants used in traditional medicine to cure infectious diseases and can be used as sources of bioactive ingredients. This current study was designed to evaluate the antibacterial activity of eight plants used in the traditional pharmacopeia of Niger. The extracts were prepared by maceration using ethanol, methanol, and distilled water. The obtained extracts were screened against spp., spp., and using the microdilution method coupled with a resazurin-based assay. Phytochemical screening was performed using colorimetry, while the quantification of total polyphenols, total flavonoids, and total tannins was determined by spectrophotometry. Out of the eight plants obtained, five named , , , , and exhibited antibacterial activity with MICs ranging from 500 g/mL to 2000 g/mL. Phytochemical screening showed the presence of alkaloids, saponosides, tannins, flavonoids, terpenes/sterols, quinones, and polyphenols. The ethanolic and methanolic extracts of contained important quantities of total polyphenols, with 43.59 ± 0.15 and 41.97 ± 0.02 mg EAG/100 mg of extract, respectively. These extracts showed the highest contents of total tannins at 46.49 g/L and 45.52 g/L, respectively. For total flavonoids, the highest content was obtained with the methanolic extract of , with 3.12 ± 0.01 mg QE/100 mg of extract. These findings justify the uses of these plants in traditional medicine for the treatment of infectious diseases such as diarrhea and can be used as starting points for the development of phytodrugs against infectious diarrhea.
PubMed: 37529122
DOI: 10.1155/2023/6120255 -
Microbial Cell Factories Jan 2024The genus Eubacterium is quite diverse and includes several acetogenic strains capable of fermenting C1-substrates into valuable products. Especially, Eubacterium...
BACKGROUND
The genus Eubacterium is quite diverse and includes several acetogenic strains capable of fermenting C1-substrates into valuable products. Especially, Eubacterium limosum and closely related strains attract attention not only for their capability to ferment C1 gases and liquids, but also due to their ability to produce butyrate. Apart from its well-elucidated metabolism, E. limosum is also genetically accessible, which makes it an interesting candidate to be an industrial biocatalyst.
RESULTS
In this study, we examined genomic, phylogenetic, and physiologic features of E. limosum and the closest related species E. callanderi as well as E. maltosivorans. We sequenced the genomes of the six Eubacterium strains 'FD' (DSM 3662), 'Marburg' (DSM 3468), '2A' (DSM 2593), '11A' (DSM 2594), 'G14' (DSM 107592), and '32' (DSM 20517) and subsequently compared these with previously available genomes of the E. limosum type strain (DSM 20543) as well as the strains 'B2', 'KIST612', 'YI' (DSM 105863), and 'SA11'. This comparison revealed a close relationship between all eleven Eubacterium strains, forming three distinct clades: E. limosum, E. callanderi, and E. maltosivorans. Moreover, we identified the gene clusters responsible for methanol utilization as well as genes mediating chain elongation in all analyzed strains. Subsequent growth experiments revealed that strains of all three clades can convert methanol and produce acetate, butyrate, and hexanoate via reverse β-oxidation. Additionally, we used a harmonized electroporation protocol and successfully transformed eight of these Eubacterium strains to enable recombinant plasmid-based expression of the gene encoding the fluorescence-activating and absorption shifting tag (FAST). Engineered Eubacterium strains were verified regarding their FAST-mediated fluorescence at a single-cell level using a flow cytometry approach. Eventually, strains 'FD' (DSM 3662), '2A' (DSM 2593), '11A' (DSM 2594), and '32' (DSM 20517) were genetically engineered for the first time.
CONCLUSION
Strains of E. limosum, E. callanderi, and E. maltosivorans are outstanding candidates as biocatalysts for anaerobic C1-substrate conversion into valuable biocommodities. A large variety of strains is genetically accessible using a harmonized electroporation protocol, and FAST can serve as a reliable fluorescent reporter protein to characterize genetically engineered cells. In total eleven strains have been assigned to distinct clades, providing a clear and updated classification. Thus, the description of respective Eubacterium species has been emended, improved, aligned, and is requested to be implemented in respective databases.
Topics: Eubacterium; Metabolic Engineering; Methanol; Phylogeny; Butyrates
PubMed: 38233843
DOI: 10.1186/s12934-024-02301-8 -
Journal of Microbiology and... Jan 2024Plants contain a large number of phytochemical components, many of which are known as bioactive compounds and responsible for the expression of various pharmacological...
Polyphenol Contents, Gas Chromatography-Mass Spectrometry (GC-MS) and Antibacterial Activity of Methanol Extract and Fractions of Fruits from Ben Tre Province in Vietnam.
Plants contain a large number of phytochemical components, many of which are known as bioactive compounds and responsible for the expression of various pharmacological activities. The extract of fruit collected in Vietnam was investigated for its total phenolic and total flavonoid contents using methanol solvent and different fractions of fruits (hexane, ethyl acetate, n-butanol, and aqueous). GC-MS analysis was conducted to identify the bioactive chemical constituents occurring in the active extract. Further, the antibacterial activity was tested in vitro on bacterial isolates, namely , , and , using the disc diffusion method on tryptic soya agar (TSA) medium. The methanol extract showed high total flavonoid (82.3 ± 0.41 mg QE/g extract) and phenolic (41.0 ± 0.34 mg GAE/g extract) content. GC-MS of the methanol extract and different fractions of fruits detected 20 compounds, principally fatty alcohols, fatty acids, phenols, lipids, terpenes derivatives, and carboxylic acids derivatives. A 50 mg/ml concentration of methanol extract had the strongest antibacterial activity on , , and . Furthermore, ethyl acetate, aqueous, and n-butanol fractions inhibited and the most. The results of the present study suggested that the fruits of are rich sources of phenolic compounds that can contribute to safe and cost-effective treatments.
Topics: Polyphenols; Fruit; Plant Extracts; Methanol; Gas Chromatography-Mass Spectrometry; Staphylococcus aureus; Vietnam; 1-Butanol; Escherichia coli; Antioxidants; Anti-Bacterial Agents; Phenols; Flavonoids; Acetates
PubMed: 38282409
DOI: 10.4014/jmb.2304.04019 -
Ultrasonics Sonochemistry Dec 2023The sonochemical generation of hydrogen (H) was investigated using various water/alcohol solutions under argon (Ar) 100 % in a 300 kHz sonoreactor. Five types of...
The sonochemical generation of hydrogen (H) was investigated using various water/alcohol solutions under argon (Ar) 100 % in a 300 kHz sonoreactor. Five types of alcohols-methanol, ethanol, isopropanol, n-propanol, and n-butanol-were used at various concentrations (0 - 100 % v/v). The H generation rate in water was 0.31 μmol/min in the absence of alcohols. The H generation rate increased, peaked, and then decreased as the alcohol concentration increased. The concentrations used for the peak H generation were 5 %, 1 %, 0.5 %, 0.5 %, and 0.1 % for methanol, ethanol, isopropanol, n-propanol, and n-butanol, respectively. The highest generation rate (5.46 μmol/min) was obtained for methanol 5 % among all conditions in this study, and no H was detected for 100 % alcohol concentrations. The reason for the enhancement of the sonochemical H generation by the addition of alcohols might be due to strong scavenging effect of alcohols for sonochemically generated oxidizing radicals and vigorous reactions of alcohol molecules and their derivatives with H radicals. No significant correlations were found between the H generation rates and physicochemical properties of the alcohols in any of the data in this study. As alcohol concentration increased, the calorimetric power decreased. This indicates that the calorimetric power does not represent the degree of sonochemical reactions in the water/alcohol mixtures. The effect of oxygen (O) content in the dissolved gases on the generation of HO (representing sonochemical oxidation activity) and H (representing sonochemical reduction activity) was investigated using Ar/O mixtures for water, methanol 5 % and n-propanol 0.5 %. In water, the highest HO generation was obtained for Ar/O (50:50), which is similar to previous research results. However, the HO generation increased as the O content increased. In addition, H generation decreased as the O content increased under all liquid conditions (water, methanol, and n-propanol).
PubMed: 37924613
DOI: 10.1016/j.ultsonch.2023.106660 -
BMC Complementary Medicine and Therapies Aug 2023Scientists and medical professionals are actively striving to improve the efficacy of treatment methods for oral squamous cell carcinoma (OSCC), the most frequently...
Cytotoxic, anti-proliferative, and apoptotic evaluation of Ramalina sinensis (Ascomycota, Lecanoromycetes), lichenized fungus on oral squamous cell carcinoma cell line; in-vitro study.
BACKGROUND
Scientists and medical professionals are actively striving to improve the efficacy of treatment methods for oral squamous cell carcinoma (OSCC), the most frequently occurring cancer within the oral cavity, by exploring the potential of natural products. The active pharmacological compounds found in lichenized fungi have shown potential for aiding in cancer treatment. Recent research aims to evaluate the impact of the lichenized fungus Ramalina sinensis (R. sinensis) on the cell viability and apoptosis of OSCC cell lines, considering the anti-inflammatory and anti-cancer capabilities of lichens.
METHODS
Ramalina sinensis (Ascomycota, Lecanoromycetes) was selected for investigation of its effects on a human oral squamous cell carcinoma cell line. Acetone and methanol extracts of R. sinensis on an OSCC cell line (KB cell line, NCBI Code: C152) were investigated. Viability was assessed by MTT assay analysis, and apoptotic cells were measured using flow cytometry analysis. Scratch assay was used to assess cell migration. The chemical composition and metabolic profiling of R. sinensis were investigated.
RESULTS
The growth and multiplication of KB cells were observed to undergo a gradual but remarkable inhibition when exposed to various concentrations. Specifically, concentrations of 6.25, 12.5, 25, 50, 100, and 200 μg/mL exhibited a significant suppressive effect on the proliferation of KB cells. The inhibition of cell proliferation exhibited a statistically significant difference between the extracts obtained from acetone and methanol. Flow cytometry results show an increase in apoptosis of OSCC cells by acetone extract. R. sinensis exerted a concentration-dependent inhibitory effect on the migration of OSCC cells. The chemical composition of R. sinensis was investigated using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and 33 compounds in the acetone and methanol extracts of R. sinensis were detected.
CONCLUSION
The findings provide evidence supporting the beneficial effects of R. sinensis extract on inducing apoptosis in OSCC cells and exerting anti-cancer properties.
Topics: Humans; Carcinoma, Squamous Cell; Squamous Cell Carcinoma of Head and Neck; Acetone; Methanol; Tandem Mass Spectrometry; Mouth Neoplasms; Ascomycota; Antineoplastic Agents; Cell Line; Head and Neck Neoplasms; Plant Extracts
PubMed: 37608377
DOI: 10.1186/s12906-023-04118-1 -
MethodsX Dec 2023Since the beginning of the COVID-19 pandemic, the use and manufacture of alcohol-based hand sanitizers increased exponentially. Efficacy of hand sanitizers mainly...
Since the beginning of the COVID-19 pandemic, the use and manufacture of alcohol-based hand sanitizers increased exponentially. Efficacy of hand sanitizers mainly depends on active ingredients like ethanol and isopropanol (IPA). Even though methanol is extremely hazardous to people, it is still illegally used in hand sanitizers in Bangladesh. Developing a quick and simple analytical method for detecting and quantifying ethanol/IPA/methanol is crucial. Here, Fourier transform infrared spectroscopy (FTIR) was used to identify and quantify alcohol content in commercially available hand sanitizers in a quick and easy way. Comparing the FTIR and GC data, provided quite similar results. Unlike previous studies by FTIR, C-H, CH-C-CH stretching, and C-H bending vibrational modes were employed to construct analytical calibration curves to detect and quantify alcohol in hand sanitizers. According to FTIR and GC findings, ethanol and IPA content were found to be 43-82% and 40-69%, and 56-64% and 61-66%, respectively, whereas ethanol was labeled at 66-80% and IPA at 65-70%. FTIR and GC revealed methanol content ranging from 37 to 98 and 19 to 81%, respectively. Also, the FTIR was significantly faster than the GC. Therefore, FTIR can be used to commercially analyze the quality of hand sanitizers.•FTIR was used to identify and quantify alcohol content in commercially available hand sanitizers in a quick and easy way.•Comparing the FTIR and GC data, provided quite similar results.•Out of ten samples, five contained ethanol, three IPA, and two methanol.
PubMed: 37484519
DOI: 10.1016/j.mex.2023.102274 -
Journal of Translational Medicine Jan 2024Hepatocellular carcinoma (HCC) remains a leading life-threatening health challenge worldwide, with pressing needs for novel therapeutic strategies. Sphingosine kinase 1...
BACKGROUND
Hepatocellular carcinoma (HCC) remains a leading life-threatening health challenge worldwide, with pressing needs for novel therapeutic strategies. Sphingosine kinase 1 (SphK1), a well-established pro-cancer enzyme, is aberrantly overexpressed in a multitude of malignancies, including HCC. Our previous research has shown that genetic ablation of Sphk1 mitigates HCC progression in mice. Therefore, the development of PF-543, a highly selective SphK1 inhibitor, opens a new avenue for HCC treatment. However, the anti-cancer efficacy of PF-543 has not yet been investigated in primary cancer models in vivo, thereby limiting its further translation.
METHODS
Building upon the identification of the active form of SphK1 as a viable therapeutic target in human HCC specimens, we assessed the capacity of PF-543 in suppressing tumor progression using a diethylnitrosamine-induced mouse model of primary HCC. We further delineated its underlying mechanisms in both HCC and endothelial cells. Key findings were validated in Sphk1 knockout mice and lentiviral-mediated SphK1 knockdown cells.
RESULTS
SphK1 activity was found to be elevated in human HCC tissues. Administration of PF-543 effectively abrogated hepatic SphK1 activity and significantly suppressed HCC progression in diethylnitrosamine-treated mice. The primary mechanism of action was through the inhibition of tumor neovascularization, as PF-543 disrupted endothelial cell angiogenesis even in a pro-angiogenic milieu. Mechanistically, PF-543 induced proteasomal degradation of the critical glycolytic enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3, thus restricting the energy supply essential for tumor angiogenesis. These effects of PF-543 could be reversed upon S1P supplementation in an S1P receptor-dependent manner.
CONCLUSIONS
This study provides the first in vivo evidence supporting the potential of PF-543 as an effective anti-HCC agent. It also uncovers previously undescribed links between the pro-cancer, pro-angiogenic and pro-glycolytic roles of the SphK1/S1P/S1P receptor axis. Importantly, unlike conventional anti-HCC drugs that target individual pro-angiogenic drivers, PF-543 impairs the PFKFB3-dictated glycolytic energy engine that fuels tumor angiogenesis, representing a novel and potentially safer therapeutic strategy for HCC.
Topics: Animals; Humans; Mice; Angiogenesis; Carcinoma, Hepatocellular; Diethylnitrosamine; Endothelial Cells; Liver Neoplasms; Methanol; Neovascularization, Pathologic; Phosphofructokinase-2; Phosphotransferases (Alcohol Group Acceptor); Pyrrolidines; Sphingosine-1-Phosphate Receptors; Sulfones
PubMed: 38200582
DOI: 10.1186/s12967-023-04830-z