acetyl-CoA Synthase Research Articles

Untargeted metabolomics to analyze alterations in two-dimensional graphdiyne–copper nanocomposite on the metabolic reprogramming of prostate cancer

Prostate cancer (PCa) is a common type of cancer in men worldwide. Metabolic reprogramming is an important factor in its pathogenesis. Two-dimensional (2D) nanomaterials have tremendous potential for cancer treatment owing to their unique physicochemical properties. However, very few studies have focused on the metabolic reprogramming mechanisms of PCa using 2D nanomaterials. In this study, for the first time, 2D graphdiyne (GDY) was used as a template to immobilize copper (Cu) ions to form a novel nanocomposite (GDY–Cu). GDY provides a large π-conjugated architecture that spatiotemporally restricts Cu ions spatiotemporally to realize the functional moiety of Cu ions as tumor therapeutics. The GDY–Cu nanocomposite with a 2D morphological structure was characterized using a transmission electron microscopy (TEM) and atomic force microscopy. The distribution of Cu loaded on the GDY was confirmed by high-resolution TEM with energy-dispersive x-ray spectroscopy analysis. In vitro and in vivo, GDY–Cu exhibits a good antitumor effect and is associated with specific metabolic reprogramming characteristics of PCa. In this study, the effect of GDY–Cu on the metabolic reprogramming of PCa cells was analyzed using untargeted metabolomics. Differences in metabolites in DU145 cells treated with GDY–Cu were analyzed by clustering and target analysis using bioinformatics methods. GDY–Cu inhibited the growth of PCa cells by decreasing the expression levels of acetyl-CoA carboxylase and cytoplasmic acetyl-CoA synthase, which inhibits the synthesis of related fatty acids and lipid metabolism. These results indicated that GDY–Cu inhibits the growth of PCa cells mainly via lipid metabolic pathways. At present, combinatory therapeutic modalities based on GDY and Cu are in their infancy. Further exploration is required to promote the development of 2D nanocomposite combinatory therapeutic modalities based on metabolic reprogramming.

Enhancing xylanase expression by Komagataella phaffii by formate as carbon source and inducer

Komagataella phaffii (syn. Pichia pastoris), a methylotrophic yeast, has many advantages as a protein expression system, but has the disadvantage of hazardous methanol as an inducer and carbon source. To enable substitution of formate for methanol, a formate assimilation pathway was constructed by the co-expression of acetyl-CoA synthase, acetaldehyde dehydrogenase, and transcription factor Mit1, resulting in a 103.5 ± 12.5% increase in xylanase production. Recombinant K. phaffii was able to use formate as a carbon source, indicating successful substitution of formate for methanol. Xylanase production, using the safe and sustainable formate as an inducer and carbon source, is a major advance in the field of industrial enzyme production. KEY POINTS: • Change to formate assimilation by recombinant K. phaffii instead of methanol • K. phaffii expressed xylanase by formate induction instead of methanol induction • Increased xylanase expression by transcription factor co-expression.

Structural Insights into Microbial One-Carbon Metabolic Enzymes Ni-Fe-S-Dependent Carbon Monoxide Dehydrogenases and Acetyl-CoA Synthases.

Ni-Fe-S-dependent carbon monoxide dehydrogenases (CODHs) are enzymes that interconvert CO and CO2 by using their catalytic Ni-Fe-S C-cluster and their Fe-S B- and D-clusters for electron transfer. CODHs are important in the microbiota of animals such as humans, ruminants, and termites because they can facilitate the use of CO and CO2 as carbon sources and serve to maintain redox homeostasis. The bifunctional carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) is responsible for acetate production via the Wood-Ljungdahl pathway, where acetyl-CoA is assembled from two CO2-derived one-carbon units. A Ni-Fe-S A-cluster is key to this chemistry. Whereas acetogens use the A- and C-clusters of CODH/ACS to produce acetate from CO2, methanogens use A- and C-clusters of an acetyl-CoA decarbonylase/synthase complex (ACDS) to break down acetate en route to CO2 and methane production. Here we review some of the recent advances in understanding the structure and mechanism of CODHs, CODH/ACSs, and ACDSs, their unusual metallocofactors, and their unique metabolic roles in the human gut and elsewhere.

The multiple effects of REG1 deletion and SNF1 overexpression improved the production of S-adenosyl-l-methionine in Saccharomyces cerevisiae

BackgroundSaccharomyces cerevisiae is often used as a cell factory for the production of S-adenosyl-l-methionine (SAM) for diverse pharmaceutical applications. However, SAM production by S. cerevisiae is negatively influenced by glucose repression, which is regulated by a serine/threonine kinase SNF1 complex. Here, a strategy of alleviating glucose repression by deleting REG1 (encodes the regulatory subunit of protein phosphatase 1) and overexpressing SNF1 (encodes the catalytic subunit of the SNF1 complex) was applied to improve SAM production in S. cerevisiae. SAM production, growth conditions, glucose consumption, ethanol accumulation, lifespan, glycolysis and amino acid metabolism were analyzed in the mutant strains.ResultsThe results showed that the multiple effects of REG1 deletion and/or SNF1 overexpression exhibited a great potential for improving the SAM production in yeast. Enhanced the expression levels of genes involved in glucose transport and glycolysis, which improved the glucose utilization and then elevated the levels of glycolytic intermediates. The expression levels of ACS1 (encoding acetyl-CoA synthase I) and ALD6 (encoding aldehyde dehydrogenase), and the activity of alcohol dehydrogenase II (ADH2) were enhanced especially in the presence of excessive glucose levels, which probably promoted the conversion of ethanol in fermentation broth into acetyl-CoA. The gene expressions involved in sulfur-containing amino acids were also enhanced for the precursor amino acid biosynthesis. In addition, the lifespan of yeast was extended by REG1 deletion and/or SNF1 overexpression. As expected, the final SAM yield of the mutant YREG1ΔPSNF1 reached 8.28 g/L in a 10-L fermenter, which was 51.6% higher than the yield of the parent strain S. cerevisiae CGMCC 2842.ConclusionThis study showed that the multiple effects of REG1 deletion and SNF1 overexpression improved SAM production in S. cerevisiae, providing new insight into the application of the SNF1 complex to abolish glucose repression and redirect carbon flux to nonethanol products in S. cerevisiae.

Development of Hypertolerant Strain of Yarrowia lipolytica Accumulating Succinic Acid Using High Levels of Acetate.

Acetate is emerging as a promising feedstock for biorefineries as it can serve as an alternate carbon source for microbial cell factories. In this study, we expressed acetyl-CoA synthase in Yarrowia lipolytica PSA02004PP, and the recombinant strain grew on acetate as the sole carbon source and accumulated succinic acid or succinate (SA). Unlike traditional feedstocks, acetate is a toxic substrate for microorganisms; therefore, the recombinant strain was further subjected to adaptive laboratory evolution to alleviate toxicity and improve tolerance against acetate. At high acetate concentrations, the adapted strain Y. lipolytica ACS 5.0 grew rapidly and accumulated lipids and SA. Bioreactor cultivation of ACS 5.0 with 22.5 g/L acetate in a batch mode resulted in a maximum cell OD600 of 9.2, with lipid and SA accumulation being 0.84 and 5.1 g/L, respectively. However, its fed-batch cultivation yielded a cell OD600 of 23.5, SA titer of 6.5 g/L, and lipid production of 1.5 g/L with an acetate uptake rate of 0.2 g/L h, about 2.86 times higher than the parent strain. Cofermentation of acetate and glucose significantly enhanced the SA titer and lipid accumulation to 12.2 and 1.8 g/L, respectively, with marginal increment in cell growth (OD600: 26.7). Furthermore, metabolic flux analysis has drawn insights into utilizing acetate for the production of metabolites that are downstream to acetyl-CoA. To the best of our knowledge, this is the first report on SA production from acetate by Y. lipolytica and demonstrates a path for direct valorization of sugar-rich biomass hydrolysates with elevated acetate levels to SA.

Dietary selection of metabolically distinct microorganisms drives hydrogen metabolism in ruminants

Ruminants are important for global food security but emit the greenhouse gas methane. Rumen microorganisms break down complex carbohydrates to produce volatile fatty acids and molecular hydrogen. This hydrogen is mainly converted into methane by archaea, but can also be used by hydrogenotrophic acetogenic and respiratory bacteria to produce useful metabolites. A better mechanistic understanding is needed on how dietary carbohydrates influence hydrogen metabolism and methanogenesis. We profiled the composition, metabolic pathways, and activities of rumen microbiota in 24 beef cattle adapted to either fiber-rich or starch-rich diets. The fiber-rich diet selected for fibrolytic bacteria and methanogens resulting in increased fiber utilization, while the starch-rich diet selected for amylolytic bacteria and lactate utilizers, allowing the maintenance of a healthy rumen and decreasing methane production (p < 0.05). Furthermore, the fiber-rich diet enriched for hydrogenotrophic methanogens and acetogens leading to increased electron-bifurcating [FeFe]-hydrogenases, methanogenic [NiFe]- and [Fe]-hydrogenases and acetyl-CoA synthase, with lower dissolved hydrogen (42%, p < 0.001). In contrast, the starch-rich diet enriched for respiratory hydrogenotrophs with greater hydrogen-producing group B [FeFe]-hydrogenases and respiratory group 1d [NiFe]-hydrogenases. Parallel in vitro experiments showed that the fiber-rich selected microbiome enhanced acetate and butyrate production while decreasing methane production (p < 0.05), suggesting that the enriched hydrogenotrophic acetogens converted some hydrogen that would otherwise be used by methanogenesis. These insights into hydrogen metabolism and methanogenesis improve understanding of energy harvesting strategies, healthy rumen maintenance, and methane mitigation in ruminants.

Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris.

Production of 3-hydroxypropionic acid (3-HP) in Pichia pastoris (syn. Komagataella phaffii) via the malonyl-CoA pathway has been recently demonstrated using glycerol as a carbon source, but the reported metrics were not commercially relevant. The flux through the heterologous pathway from malonyl-CoA to 3-HP was hypothesized as the main bottleneck. In the present study, different metabolic engineering approaches have been combined to improve the productivity of the original 3-HP producing strains. To do so, an additional copy of the gene encoding for the potential rate-limiting step of the pathway, i.e., the C-terminal domain of the malonyl-CoA reductase, was introduced. In addition, a variant of the endogenous acetyl-CoA carboxylase (ACC1 S1132A ) was overexpressed with the aim to increase the delivery of malonyl-CoA. Furthermore, the genes encoding for the pyruvate decarboxylase, aldehyde dehydrogenase and acetyl-CoA synthase, respectively, were overexpressed to enhance conversion of pyruvate into cytosolic acetyl-CoA, and the main gene responsible for the production of the by-product D-arabitol was deleted. Three different screening conditions were used to classify the performance of the different strains: 24-deep-well plates batch cultures, small-scale cultures in falcon tubes using FeedBeads® (i.e., slow release of glycerol over time), and mini bioreactor batch cultures. The best two strains from the FeedBeads® screening, PpHP8 and PpHP18, were tested in bioreactor fed-batch cultures using a pre-fixed exponentially increasing feeding rate. The strain PpHP18 produced up to 37.05 g L−1 of 3-HP at 0.712 g L−1 h−1 with a final product yield on glycerol of 0.194 Cmol−1 in fed-batch cultures. Remarkably, PpHP18 did not rank among the 2-top producer strains in small scale batch cultivations in deep-well plates and mini bioreactors, highlighting the importance of multiplexed screening conditions for adequate assessment of metabolic engineering strategies. These results represent a 50% increase in the product yield and final concentration, as well as over 30% increase in volumetric productivity compared to the previously obtained metrics for P. pastoris. Overall, the combination of glycerol as carbon source and a metabolically engineered P. pastoris strain resulted in the highest 3-HP concentration and productivity reported so far in yeast.

Evaluation of the synergistic effects of biochar and biogas residue on CO2 and CH4 emission, functional genes, and enzyme activity during straw composting

This study examined the effects of biochar, biogas residue, and their combined amendments on CO2 and CH4 emission, enzyme activity, and related functional genes during rice straw composting. Results showed that the biogas residue increased CO2 and CH4 emissions by 13.07 % and 74.65 %, while biochar had more obvious inhibition. Biogas residue addition enhanced functional gene abundance more than biochar. Biogas residue raised the methanogens mcrA gene by 2.5 times. Biochar improved the Acetyl-CoA synthase and β-glucosidase activities related to carbon fixation and decreased coenzyme activities related to methanogens. Biochar and biogas residue combined amendments enhanced the acsB gene abundance for CO2 assimilation process and decreased methyl-coenzyme M reductase α subunit activity. Pearson correlation analysis indicated that organic matter was the significant variable affecting CO2 and CH4 emissions (P < 0.01). These results indicated biochar played significant roles in carbon loss and greenhouse emissions caused by biogas residue incorporation during composting.

Size-dependent effects of polystyrene microplastics on anaerobic digestion performance of food waste: Focusing on oxidative stress, microbial community, key metabolic functions

Polystyrene (PS) microplastics (MPs) are widely existed in food waste (FW) due to the usage of plastic food-packaging. However, the effects and mechanisms of PS MPs with different sizes on anaerobic digestion (AD) performance of FW have not been comprehensively studied yet. Herein, the impacts of different PS MPs sizes (1 mm, 100 µm and 1 µm) with 20, 200 particles/g-TS were investigated. Results showed that 20 particles/g-TS PS MPs decreased cumulative methane production by 1.46–18.11 %, while the higher levels (200 particles/g-TS) significantly inhibited by 9.14–33.08 % (p < 0.05) compared with control group. The inhibiting effects were enhanced as particle size smaller. Physicochemical analysis indicated that MPs prolonged organic matter hydrolysis, weakened the volatile fatty acids metabolism and inhibited methanogenesis-related microorganisms (Synergistetes, Proteiniphilum and Methanosarcina). Small-sized MPs could induce more reactive oxygen species causing cell toxicity and suppressed key enzymes (α–glucoside, protease, acetate kinases and F420) activities, thereby restraining methane production. The analyses of acetyl-CoA synthase and methyl-coenzyme M reductase functional genes illustrated that small-sized MPs negatively affected acetoclastic methanogenesis pathways. Overall, these results provide new insights into the size-dependent effects on AD performance induced by PS MPs.

Development of CO gas conversion system using high CO tolerance biocatalyst

Acetogenic bacteria (acetogens) are microorganisms with high potential as biocatalysts that can convert C1 gas, such as carbon dioxide (CO2) or carbon monoxide (CO), into biomass or biochemicals; however, high CO concentrations cause growth retardation in these microorganisms. In this study, we performed adaptive evolution of Eubacterium limosum, an acetogen, for approximately 400 generations under high CO synthetic gas (syngas) conditions to obtain E. limosum ECO2, which can rapidly proliferate even under high CO conditions. A mutation was revealed in H636R in the Acetyl-CoA synthase (ACS) protein of the ECO2 strain through whole-genome re-sequencing, and the mutation increased the CO gas conversion rate. By analyzing the transcript profiling of the ECO2 strain under high CO syngas conditions, the transcript levels of the methyl branch in the Wood–Ljungdahl pathway and the energy conservation system increased at high CO concentration conditions. Finally, to convert CO into value-added biochemicals, 2,3-BDO (2,3-butanediol) biosynthesis pathways were constructed and transformed into the E. limosum ECO2 strain. This strain showed capability to produce 2.60 mg L-1h−1 of 2,3-BDO through CO 66% syngas fed-batch fermentation. This was 6.5-fold higher than that of the wild type. We developed a microorganism with high CO tolerance, and this strain showed high application potential as a biocatalyst that can efficiently convert CO gas into value-added chemicals.

Key Molecules of Fatty Acid Metabolism in Gastric Cancer.

Fatty acid metabolism is closely linked to the progression of gastric cancer (GC), a very aggressive and life-threatening tumor. This study examines linked molecules, such as Sterol Regulatory Element-Binding Protein 1 (SREBP1), ATP Citrate Lyase (ACLY), Acetyl-CoA Synthases (ACSs), Acetyl-CoA Carboxylase (ACC), Fatty Acid Synthase (FASN), Stearoyl-CoA Desaturase 1 (SCD1), CD36, Fatty Acid Binding Proteins (FABPs), and Carnitine palmitoyltransferase 1 (CPT1), as well as their latest studies and findings in gastric cancer to unveil its core mechanism. The major enzymes of fatty acid de novo synthesis are ACLY, ACSs, ACC, FASN, and SCD1, while SREBP1 is the upstream molecule of fatty acid anabolism. Fatty acid absorption is mediated by CD36 and FABPs, and fatty acid catabolism is mediated by CPT1. If at all possible, we will discover novel links between fatty acid metabolism and a prospective gastric cancer target.

Multiple resistance of silky windgrass to acetolactate synthase- and acetyl-CoA synthase–inhibiting herbicides

AbstractField and pot experiments were conducted in Greece to study the occurrence of resistance in silky windgrass to acetolactate synthase (ALS)- and acetyl-CoA synthase (ACCase)-inhibiting herbicides. Twenty-four populations of silky windgrass were examined in whole-plant response experiments. High levels of field-evolved resistance to chlorsulfuron (0% to 28% control in terms of fresh weight reduction) with the recommended field rates were confirmed in most silky windgrass populations. However, other ALS inhibitors, such as pyroxsulam and a premix of mesosulfuron-methyl and iodosulfuron, provided adequate control (76% to 100% in terms of fresh weight reduction) of most populations, except eight silky windgrass populations that were found to be cross-resistant to all ALS-inhibiting herbicides tested (i.e., chlorsulfuron, commercial mixture of mesosulfuron-methyl plus iodosulfuron, and pyroxsulam). Conversely, most silky windgrass populations were controlled effectively (90% to 100% in terms of fresh weight reduction) with the recommended field rates of ACCase inhibitors cycloxydim, clethodim, and pinoxaden, but five populations were also found to be resistant to clodinafop-propargyl (10% to 68% control in terms of fresh weight reduction). The ALS gene sequencing of the eight silky windgrass populations, with cross-resistance to ALS inhibitors, revealed a point mutation at the Pro-197 position, causing amino acid substitution by Ser or Thr in the ALS enzyme. Overall, chlorsulfuron and clodinafop-propargyl were selecting agents of field-evolved multiple resistance to ALS- and ACCase-inhibiting herbicides in five silky windgrass populations. As the available postemergence-applied chemistries/modes of action registered for grass weed control in cereals are rather limited, adopting integrated management practices and implementing proactive and reactive measures to delay the evolution of resistant populations is essential.

Exploitation of strong constitutive and stress-driven promoters from Acetobacter pasteurianus for improving acetic acid tolerance

Acetobacter pasteurianus is an excellent cell factory for production of highly-strength acetic acid, and attracts an increasing attention in metabolic engineering. However, the available well-characterized constitutive and inducible promoters are rather limited to adjust metabolic fluxes in A. pasteurianus. In this study, we screened a panel of constitutive and acid stress-driven promoters based on time-series of RNA-seq data and characterized in A. pasteurianus and Escherichia coli. Nine constitutive promoters ranged in strength from 1.7-fold to 100-fold that of the well-known strong promoter Padh under non-acetic acid environment. Subsequently, an acetic acid-stable red fluorescent visual reporting system was established and applied to evaluate acid stress-driven promoter in A. pasteurianus during highly-acidic fermentation environment. PgroES was identified as acid stress-driven strong promoters, with expression outputs varied from 100% to 200% when acetic acid treatment. To assess their application potential, ultra-strong constitutive promoter Ptuf and acid stress-driven strong promoter PgroES were selected to overexpress acetyl-CoA synthase and greatly improved acetic acid tolerance. Notably, the acid stress-driven promoter displayed more favorable for regulating strain robustness against acid stress by overexpressing tolerance gene. In summary, this is the first well-characterized constitutive and acid stress-driven promoter library from A. pasteurianus, which could be used as a promising toolbox for metabolic engineering in acetic acid bacteria and other gram-negative bacteria.

Influence of Nutrient (NPK) Factors on Growth, and Pharmacodynamic Component Biosynthesis of Atractylodes chinensis: An Insight on Acetyl-CoA Carboxylase (ACC), 3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGR), and Farnesyl Pyrophosphate Synthase (FPPS) Signaling Responses.

In the planting of crops, especially medicinal plants, formula fertilization is important for improving the utilization rate of elements, soil quality, crop yield, and quality. Therefore, it is important to study targeted fertilizer application schemes for sustainable agricultural development and environmental protection. In this study, an L9(34) orthogonal design was used to conduct a field experiment to study the effects of NPK combined application on the growth and pharmacodynamic component biosynthesis of Atractylodes chinensis (DC.) Koidz. Results showed that after applying a base fertilizer at the seedling stage (late May), topdressing at the vegetative stage (late June) and fruit stage (late August) was beneficial to the growth and development of A. chinensis. The high concentrations of phosphorus were conducive to the accumulation of yield and effective components, and the best harvest time was after late October. Principal component analysis (PCA) showed that the comprehensive score of T6 treatment was the highest, indicating that the optimal fertilization scheme for the high yield and high quality of A. chinensis was (N2P3K1): N 180, P2O5 225, and K2O 105 kg⋅ha–1. A signaling response analysis showed that during the growth and development of A. chinensis, the T6 fertilization scheme had clear effects on the activity and gene expression of the key enzymes acetyl-CoA carboxylase (ACC) and farnesyl pyrophosphate synthase (FPPS). Under the T4 [(N2P1K2): N 180, P2O5 75, and K2O 210 kg⋅ha–1] fertilization scheme, the activity and gene expression of the key enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) were higher. Moreover, ACC was closely related to the synthesis of the polyacetylene component atractylodin, and FPPS played an important regulatory role in the synthesis of sesquiterpene components atractylenolide II, β-eudesmol, and atractylon. In summary, the high phosphorus fertilization scheme T6 could notably increase the yield of A. chinensis, and promote the accumulation of polyacetylene and sesquiterpene volatile oils by increasing the expression of ACC and FPPS. Therefore, we postulate that the precise application of nutrients (NPK) plays a vital role in the yield formation and quality regulation of A. chinensis.

Effects of Culture Mechanism of Cinnamomum kanehirae and C. camphora on the Expression of Genes Related to Terpene Biosynthesis in Antrodia cinnamomea

The rare edible and medicinal fungus Antrodia cinnamomea has a substantial potential for development. In this study, Illumina HiSeq 2000 was used to sequence its transcriptome. The results were assembled de novo, and 66,589 unigenes with an N50 of 4413 bp were obtained. Compared with public databases, 6,061, 3,257, and 2,807 unigenes were annotated to the Non-Redundant, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes databases, respectively. The genes related to terpene biosynthesis in the mycelia of A. cinnamomea were analyzed, and acetyl CoA synthase (ACS2 and ACS4), hydroxymethylglutaryl CoA reductase (HMGR), farnesyl transferase (FTase), and squalene synthase (SQS) were found to be upregulated in XZJ (twig of C. camphora) and NZJ (twig of C. kanehirae). Moreover, ACS5 and 2,3-oxidized squalene cyclase (OCS) were highly expressed in NZJ, while heme IX farnesyl transferase (IX-FIT) and ACS3 were significantly expressed in XZJ. The differential expression of ACS1, ACS2, HMGR, IX-FIT, SQS, and OCS was confirmed by real-time quantitative reverse transcription PCR. This study provides a new concept for the additional exploration of the molecular regulatory mechanism of terpenoid biosynthesis and data for the biotechnology of terpenoid production.

Acetyl CoA synthase 2 potentiates ATG5-induced autophagy against neuronal apoptosis after subarachnoid hemorrhage.

ATG5-induced autophagy is triggered in the early stages after SAH, which plays a vital role in subarachnoid hemorrhage (SAH). Acyl-CoA synthetase short-chain family 2 (ACSS2) is not just involved in energy metabolism but also binds to TEFB to form a complex translocated to related autophagy genes to regulate the expression of autophagy-related genes. However, the contribution of ACSS2 to the activation of autophagy in early brain injury (EBI) after SAH has barely been discussed. The purpose of this study was to investigate the alterations of ACSS2 and its neuroprotective effects following SAH. We first evaluated the expression of ACSS2 at different time points (6, 12, 24, and 72 h after SAH) in vivo and primary cortical neurons stimulated by oxyhemoglobin (OxyHb). Subsequently, adeno-associated virus and lentivirus were used to regulate ACSS2 expression to investigate the effect of ACSS2 after SAH. The results showed that the ACSS2 level decreased significantly in the early stages of SAH and was minimized at 24 h post-SAH. After artificial intervention to overexpress ACSS2, ATG5-induced autophagy was further enhanced in EBI after SAH, and neuronal apoptosis was alleviated to protect brain injury. In addition, brain edema and neurological function scores were improved. These results suggest that ACSS2 plays an important role in the neuroprotection against EBI after SAH by increasing ATG5-induce autophagy and inhibiting apoptosis.

Theoretical Studies of Acetyl-CoA Synthase Catalytic Mechanism

DFT calculations were performed for the A-cluster from the enzyme Acetyl-CoA synthase (ACS). The acid constants (pKa), reduction potentials, and pH-dependent reduction potential for the A-cluster with different oxidation states and ligands were calculated. Good agreement of the reduction potentials, dependent on pH in the experiment, was obtained. On the basis of the calculations, a mechanism for the methylation reaction involving two–electron reduction and protonation on the proximal nickel atom of the reduced A-cluster is proposed.

The monofunctional CO dehydrogenase CooS is essential for growth of Thermoanaerobacter kivui on carbon monoxide

Thermoanaerobacter kivui is a thermophilic acetogen that can grow on carbon monoxide as sole carbon and energy source. To identify the gene(s) involved in CO oxidation, the genome sequence was analyzed. Two genes potentially encoding CO dehydrogenases were identified. One, cooS, potentially encodes a monofunctional CO dehydrogenase, whereas another, acsA, potentially encodes the CODH component of the CODH/ACS complex. Both genes were cloned, a His-tag encoding sequence was added, and the proteins were produced from a plasmid in T. kivui. His-AcsA copurified by affinity chromatography with AcsB, the acetyl-CoA synthase of the CO dehydrogenase/acetyl CoA synthase complex. His-CooS copurified with CooF1, a small iron-sulfur center containing protein likely involved in electron transport. Both protein complexes had CO:ferredoxin oxidoreductase as well as CO:methyl viologen oxidoreductase activity, but the activity of CooSF1 was 15-times and 231-times lower, respectively. To underline the importance of CooS, the gene was deleted in the CO-adapted strain. Interestingly, the ∆cooS deletion mutant did not grow on CO anymore. These experiments clearly demonstrated that CooS is essential for growth of T. kivui on CO. This is in line with the hypothesis that CooS is the CO-oxidizing enzyme in cells growing on CO.

Altered synthesis of genes associated with short-chain fatty acids in the gut of patients with atrial fibrillation

BackgroundThe gut microbiota provides health benefits in humans by producing short-chain fatty acids (SCFAs), whose deficiency causes multiple disorders and inflammatory diseases. However, gut bacteria producing SCFAs in patients with atrial fibrillation (AF), an arrhythmia with increasing prevalence, have not been reported. To investigate major gut microbial organisms related to SCFA synthesis, SCFAs-associated KEGG orthologues (KOs), enzymatic genes, and potential producers were examined according to metagenomic data-mining in a northern Chinese cohort comprising 50 non-AF control and 50 AF patients.ResultsCompared with non-AF controls, individuals with AF had marked differences in microbial genes involved in SCFA-related synthesis, including 125 KOs and 5 SCFAs-related enzymatic genes. Furthermore, there were 10 species that harbored SCFA-synthesis related enzymatic genes, and were markedly decreased in the gut of AF patients. Notably, discriminative features about SCFA-synthesis related function, including 8 KOs (K01752, K01738, K00175, K03737, K01006, K01653, K01647 and K15023), 4 genes (menI, tesB, yciA and CO dehydrogenase acetyl-CoA synthase complex) and 2 species (Coprococcus catus and Firmicutes bacterium CAG:103), were selected as key factors based on LASSO analysis. Furthermore, PLS-SEM analysis showed that 72.8 and 91.14 % of the overall effects on gut microbiota diversity and key species on AF, respectively, were mediated by the key KOs. Meanwhile, 46.31 % of the total effects of SCFA-synthesis related function on left atrial enlargement was mediated by hsCRP. Upon incorporation of clinical properties in AF, the KO score was still significantly associated with AF incidence (OR = 0.004, P = 0.001).ConclusionsThe current study revealed that dysbiotic gut microbiota in AF is coupled with disrupted SCFA-synthesis related genes, characterized by decreased abundances of KEGG orthologues, synthesis enzymatic genes and harboring species.

Polymorphism in acetyl-CoA synthase mimic complex [NiN2S2-(W(CO)5)2

One trimetallic compound (μ-η2-Ni-1)[W(CO)5]2 (NiW2) was prepared by reaction of the Ni(II) dithiolate metalloligand, [Ni-bme-dach] where bme-dach = bis-(mercaptoethyl)diazacycloheptane, with W(CO)6 in CH2Cl2 under a certain treatment. Two solvent-free polymorphs of NiW2 (I and II) and two solvates NiW2(CH2Cl2) (III) and NiW2(C6H6) (IV) were obtained by crystallization in various solvent conditions. Conformations I and II exhibit concomitant polymorphisms in CH2Cl2 solvent system. However, conformations of III and IV were obtained in mixed solvent system of CH2Cl2 with CHCl3 and benzene, respectively. For morph II*, it was obtained by heating treatment of conformation II at 160 °C for 20 min. Two low-valent metallic W0(CO)5 groups in I to IV were bridged by Ni-1 in the transoid modes, giving W-W distances of 5.576(17) Å to 6.024(3) Å.