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CO2 fixation plays a key role to make biobased production cost competitive. Here, we use 3-hydroxypropionic acid (3-HP) to showcase how CO2 fixation enables approaching theoretical-yield production. Using genome-scale metabolic models to calculate the production envelope, we demonstrate that the provision of bicarbonate, formed from CO2, restricts previous attempts for high yield production of 3-HP. We thus develop multiple strategies for bicarbonate uptake, including the identification of Sul1 as a potential bicarbonate transporter, domain swapping of malonyl-CoA reductase, identification of Esbp6 as a potential 3-HP exporter, and deletion of Uga1 to prevent 3-HP degradation. The combined rational engineering increases 3-HP production from 0.14 g/L to 11.25 g/L in shake flask using 20 g/L glucose, approaching the maximum theoretical yield with concurrent biomass formation. The engineered yeast forms the basis for commercialization of bio-acrylic acid, while our CO2 fixation strategies pave the way for CO2 being used as the sole carbon source. CO2 fixation plays an important role to make bioproduction cost competitive. Here, the authors take 3-hydroxypropionic acid as an example to showcase how to achieve high carbon yield production through increasing the accessible bicarbonate, minimizing native CO2 release and avoiding carbon waste. [ABSTRACT FROM AUTHOR]

The growth status, root exudates, radial oxygen loss, and sewage purification effect of plants acclimated with sewage and cultured with a nutrient solution were studied by selecting Acorus gramineus Aiton, Pistia stratiotes L., and Eichhornia crassipes (Mart.) Solms with the same individual size. The results showed that the root oxygen secretion rate of the Acorus gramineus Aiton was higher in light and dark conditions. For a single plant species, the oxygen secretion rate under light conditions was much higher than that under dark conditions. The root oxygen secretion rate of Acorus gramineus Aiton was the highest (2.03 µmol O2/h/plant), followed by Pistia stratiotes L. (1.68 µmol O2/h/plant), and the root oxygen secretion rate of Eichhornia crassipes (Mart.) Solms was the lowest (1.15 µmol O2/h/plant). After a period of adaptation, plants showed strong removal effects on COD, NH3-N, TN, and TP. The removal intensity was in the order of Acorus gramineus Aiton > Pistia stratiotes L. > Eichhornia crassipes (Mart.) Solms; the higher the temperature, the bigger the removal rate. These results were consistent with Acorus gramineus Aiton's organic matter secretion and radial oxygen loss, which were better than those of Pistia stratiotes L. and Eichhornia crassipes (Mart.) Solms. Three kinds of plants had better COD removal effects in an acidic environment and better TP removal effects in an alkaline environment. The results show that using the Acorus gramineus Aiton can achieve better remediation of polluted water bodies. [ABSTRACT FROM AUTHOR]

In this study, a modified horizontal settling tank with rotating biological discs was developed to treat slightly polluted surface water, and its performance on the simultaneous removal of turbidity, organics, and NH 4 + -N was investigated on a lab scale. Results show that the effluent quality of the modified settling tank is stable in more than two months of continuous operation. At a hydraulic retention time (HRT) of 2.0 h, 73.65 ± 5.15% turbidity, 53.98 ± 5.17% TOC, and 77.01 ± 10.02%, NH 4 + -N could be removed by the modified settling tank with an average of 1.96 NTU turbidity, 1.98 mg/L TOC, and 0.46 mg/L NH 4 + -N residue in the effluent. Due to the improvement in DO supply, higher removal efficiencies of both organics and NH 4 + -N were achieved with increased disc rotating speed (r < 4 r/min). Further study showed that the genus Hyphomicrobium dominant on the posterior discs and the genus Nitrospira dominant on the anterior discs mainly contributed to the enhanced bio-oxidation of organics and NH 4 + -N, respectively. [ABSTRACT FROM AUTHOR]

Large-scale fumaric acid chemical brownfield sites have posed a significant obstacle to environmental safety, public safety, and the redevelopment of brownfield sites. A comprehensive study was conducted to determine the main pollution indicators, soil pollution characteristics, and the multiple potential risks to the environment and the ecosystem of a fumaric acid brownfield site in northwestern China. The results showed that 1,2,3-trichloropropane(1,2,3-TCP) was the primary pollutant at the fumaric acid brownfield site. The atmospheric volatile organic compounds (VOCs) in this brownfield site did not exceed the Chinese standard limits. The soil contamination was more severe within the site, with a total of about 30 VOCs detected, including the uncommon brominated pollutants. The level of longitudinal soil contamination decreased with increasing soil depth. The distillation area was more contaminated with a maximum exceedance multiple of 11,291.8. The longitudinal contamination depths in the distillation and reactor zones were 10.0 m and 4.0 m, respectively. Soil texture and production processes are considered to be one of the influencing factors for the depth of vertical soil contamination. Our findings heighten the awareness of brownfield site soil contamination and provide a reference for contamination controls and the comprehensive management of fumaric acid brownfield sites. [ABSTRACT FROM AUTHOR]

Vitamin K epoxide reductases (VKORs) are a large family of integral membrane enzymes found from bacteria to humans. Human VKOR, specific target of warfarin, has both the epoxide and quinone reductase activity to maintain the vitamin K cycle. Bacterial VKOR homologs, however, are insensitive to warfarin inhibition and are quinone reductases incapable of epoxide reduction. What affords the epoxide reductase activity in human VKOR remains unknown. Here, we show that a representative bacterial VKOR homolog can be converted to an epoxide reductase that is also inhibitable by warfarin. To generate this new activity, we first substituted several regions surrounding the active site of bacterial VKOR by those from human VKOR based on comparison of their crystal structures. Subsequent systematic substitutions narrowed down to merely eight residues, with the addition of a membrane anchor domain, that are responsible for the epoxide reductase activity. Substitutions corresponding to N80 and Y139 in human VKOR provide strong hydrogen bonding interactions to facilitate the epoxide reduction. The rest of six substitutions increase the size and change the shape of the substrate‐binding pocket, and the membrane anchor domain stabilizes this pocket while allowing certain flexibility for optimal binding of the epoxide substrate. Overall, our study reveals the structural features of the epoxide reductase activity carried out by a subset of VKOR family in the membrane environment. [ABSTRACT FROM AUTHOR]

Abstract Background Although a variety of methods and expensive kits are available, molecular cloning can be a time-consuming and frustrating process. Results Here we report a highly simplified, reliable, and efficient PCR-based cloning technique to insert any DNA fragment into a plasmid vector or into a gene (cDNA) in a vector at any desired position. With this method, the vector and insert are PCR amplified separately, with only 18 cycles, using a high fidelity DNA polymerase. The amplified insert has the ends with ~16-base overlapping with the ends of the amplified vector. After DpnI digestion of the mixture of the amplified vector and insert to eliminate the DNA templates used in PCR reactions, the mixture is directly transformed into competent E. coli cells to obtain the desired clones. This technique has many advantages over other cloning methods. First, it does not need gel purification of the PCR product or linearized vector. Second, there is no need of any cloning kit or specialized enzyme for cloning. Furthermore, with reduced number of PCR cycles, it also decreases the chance of random mutations. In addition, this method is highly effective and reproducible. Finally, since this cloning method is also sequence independent, we demonstrated that it can be used for chimera construction, insertion, and multiple mutations spanning a stretch of DNA up to 120 bp. Conclusion Our FastCloning technique provides a very simple, effective, reliable, and versatile tool for molecular cloning, chimera construction, insertion of any DNA sequences of interest and also for multiple mutations in a short stretch of a cDNA.

Accumulating epidemiological evidence supports that chronic exposure to ambient fine particular matters of <2.5 μm (PM2.5) predisposes both children and adults to Alzheimer's disease (AD) and age-related brain damage leading to dementia. There is also experimental evidence to show that PM2.5 exposure results in early onset of AD-related pathologies in transgenic AD mice and development of AD-related and age-related brain pathologies in healthy rodents. Studies have also documented that PM2.5 exposure causes AD-linked molecular and cellular alterations, such as mitochondrial dysfunction, synaptic deficits, impaired neurite growth, neuronal cell death, glial cell activation, neuroinflammation, and neurovascular dysfunction, in addition to elevated levels of amyloid β (Aβ) and tau phosphorylation. Oxidative stress and the oxidative stress-sensitive TRPM2 channel play important roles in mediating multiple molecular and cellular alterations that underpin AD-related cognitive dysfunction. Documented evidence suggests critical engagement of oxidative stress and TRPM2 channel activation in various PM2.5-induced cellular effects. Here we discuss recent studies that favor causative relationships of PM2.5 exposure to increased AD prevalence and AD- and age-related pathologies, and raise the perspective on the roles of oxidative stress and the TRPM2 channel in mediating PM2.5-induced predisposition to AD and age-related brain damage. [ABSTRACT FROM AUTHOR]

The transient receptor potential melastatin‐related 2 (TRPM2) channel, a reactive oxygen species (ROS)‐sensitive cation channel, has been well recognized for being an important and common mechanism that confers the susceptibility to ROS‐induced cell death. An elevated level of ROS is a salient feature of ischaemia‐reperfusion, chronic cerebral hypo‐perfusion and neonatal hypoxia‐ischaemia. The TRPM2 channel is expressed in hippocampus, cortex and striatum, the brain regions that are critical for cognitive functions. In this review, we examine the recent studies that combine pharmacological and/or genetic interventions with using in vitro and in vivo models to demonstrate a crucial role of the TRPM2 channel in brain damage by ischaemia‐reperfusion, chronic cerebral hypo‐perfusion and neonatal hypoxic‐ischaemia. We also discuss the current understanding of the underlying TRPM2‐dependent cellular and molecular mechanisms. These new findings lead to the hypothesis of targeting the TRPM2 channel as a potential novel therapeutic strategy to alleviate brain damage and cognitive dysfunction caused by these conditions. [ABSTRACT FROM AUTHOR]

Arsenic (As) is among the most dangerous metalloids and is harmful to human wellbeing. In this laboratory study, Al(III)-modified kapok fibres (Al-Kapok) were used to remove As(V) from water. The sorbent was characterised using Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) combined with energy-dispersive X-ray spectroscopy (EDX). Batch experiments were performed to observe the performance of Al-Kapok in the removal of As(V) and to examine the effects of pH, temperature, adsorbent dose, and coexisting ions on the adsorption process. The surface of the sorbent changed after aluminium modification, and the results of the batch experiments showed that the adsorption of As(V) occurred mainly via endothermic-spontaneous chemisorption at the solution and solid interface of Al-Kapok. The As(V) removal efficiency was approximately 76%–84%, and it was slightly affected at pH levels below 8.0. Further study showed that the maximum adsorption capacity of Al-Kapok for As(V) was 118 μg/g at 30 °C and pH 6, and notable adverse effects were caused by the presence of SO 4 2 − and PO 4 3 − . It was also found that the boundary layer and film diffusion contributed more to As(V) adsorption. After five adsorption/desorption cycles, regeneration recovered approximately 92% of the adsorption capacity of Al-Kapok used. Overall, Al-Kapok appears to be a suitable adsorbent material for the purification of As-contaminated water. [Display omitted] • The adsorption of As(V) on the Al-Kapok is slightly affected by pH at pHs below 8.0 • As(V) adsorption is mainly a chemical process and inhibited by multi-valent anions. • Boundary layer and film diffusion shows more contribution to the adsorption of As(V). • 92% of the Al-Kapok's adsorption capacity could be recovered through regeneration. [ABSTRACT FROM AUTHOR]

More than 170 million individuals have been influenced by arsenic (As) because of the ingestion of As-polluted groundwater. The presence of As in water bodies, particularly groundwater, has been found to become a widespread issue in the past few decades. Because arsenic causes extreme wellbeing impacts, even at a low concentration in drinking water, the innovations of As removal from contaminated water are of significant importance. Traditional strategies, for example, reverse osmosis, ion exchange, and electro-dialysis are generally utilized for the remediation of As-polluted water; however, the high cost and/or sludge production restricts their application in less-developed areas. The utilization of adsorbents acquired from natural materials has been explored as an alternative for the costly techniques for As removal. This paper aims to review the past and current developments in using naturals adsorbents or modified natural materials for arsenic removal and show the different parameters, which may influence the As removal effectiveness of the natural adsorbent, such as contact time, adsorbent dosage, flow rate, pH, reusability, temperature, and influence of others ions. [ABSTRACT FROM AUTHOR]

Human neuroblastoma SH-SY5Y cells are a widely-used human neuronal cell model in the study of neurodegeneration. A recent study shows that, 1-methyl-4-phenylpyridine ion (MPP), which selectively causes dopaminergic neuronal death leading to Parkinson's disease-like symptoms, can reduce SH-SY5Y cell viability by inducing H2O2 generation and subsequent TRPM2 channel activation. MPP-induced cell death is enhanced by increasing the TRPM2 expression. By contrast, increasing the TRPM2 expression has also been reported to support SH-SY5Y cell survival after exposure to H2O2, leading to the suggestion of a protective role for the TRPM2 channel. To clarify the role of reactive oxygen species (ROS)-induced TRPM2 channel activation in SH-SY5Y cells, we generated a stable SH-SY5Y cell line overexpressing the human TRPM2 channel and examined cell death and cell viability after exposure to H2O2 in the wild-type and TRPM2-overexpressing SH-SY5Y cells. Exposure to H2O2 resulted in concentration-dependent cell death and reduction in cell viability in both cell types. TRPM2 overexpression remarkably augmented H2O2-induced cell death and reduction in cell viability. Furthermore, H2O2-induced cell death in both the wild-type and TRPM2-overexpressing cells was prevented by 2-APB, a TRPM2 inhibitor, and also by PJ34 and DPQ, poly(ADP-ribose) polymerase (PARP) inhibitors. Collectively, our results show that increasing the TRPM2 expression renders SH-SY5Y cells to be more susceptible to ROS-induced cell death and reinforce the notion that the TRPM2 channel plays a critical role in conferring ROS-induced cell death. It is anticipated that SH-SY5Y cells can be useful for better understanding the molecular and signaling mechanisms for ROS-induced TRPM2-mediated neurodegeneration in the pathogenesis of neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

Although many prokaryotes have glycolysis alternatives, it's considered as the only energy-generating glucose catabolic pathway in eukaryotes. Here, we managed to create a hybrid-glycolysis yeast. Subsequently, we identified an inositol pyrophosphatase encoded by OCA5 that could regulate glycolysis and respiration by adjusting 5-diphosphoinositol 1,2,3,4,6-pentakisphosphate (5-InsP 7) levels. 5-InsP 7 levels could regulate the expression of genes involved in glycolysis and respiration, representing a global mechanism that could sense ATP levels and regulate central carbon metabolism. The hybrid-glycolysis yeast did not produce ethanol during growth under excess glucose and could produce 2.68 g/L free fatty acids, which is the highest reported production in shake flask of Saccharomyces cerevisiae. This study demonstrated the significance of hybrid-glycolysis yeast and determined Oca5 as an inositol pyrophosphatase controlling the balance between glycolysis and respiration, which may shed light on the role of inositol pyrophosphates in regulating eukaryotic metabolism. [Display omitted] • Hybrid of glycolysis with the phosphoketolase pathway made an efficient cell factory • Oca5, identified as an inositol pyrophosphatase, degrading 5-InsP 7 to InsP 6 • InsPs control gene expression involved in glycolysis and respiration to balance fluxes • The mechanism of Oca5 regulating glycolysis and respiration was elaborated Glycolysis is considered as the only energy-generating glucose catabolic pathway in eukaryotes. Qin et al. establish a hybrid-glycolysis yeast that disrupts the Embden-Meyerhof-Parnas glycolysis pathway and introduced components of the phosphoketolase pathway. They further identify Oca5 as an inositol pyrophosphatase controlling the balance between glycolysis and respiration activities. [ABSTRACT FROM AUTHOR]