Metabolic flux analysis of coenzyme Q10 synthesized by Rhodobacter sphaeroides under the influence of different pH regulators.

Coenzyme Q10 (CoQ10) is crucial for human beings, especially in the fields of biology and medicine. The aim of this experiment was to investigate the conditions for increasing CoQ10 production. At present, microbial fermentation is the main production method of CoQ10, and the production process of microbial CoQ10 metabolism control fermentation is very critical. Metabolic flux is one of the most important determinants of cell physiology in metabolic engineering. Metabolic flux analysis (MFA) is used to estimate the intracellular flux in metabolic networks. In this experiment, Rhodobacter sphaeroides was used as the research object to analyze the effects of aqueous ammonia (NH3·H2O) and calcium carbonate (CaCO3) on the metabolic flux of CoQ10. When CaCO3 was used to adjust the pH, the yield of CoQ10 was 274.43 mg·L-1 (8.71 mg·g-1 DCW), which was higher than that of NH3·H2O adjustment. The results indicated that when CaCO3 was used to adjust pH, more glucose-6-phosphate (G6P) entered the pentose phosphate (HMP) pathway and produced more NADPH, which enhanced the synthesis of CoQ10. At the chorismic acid node, more metabolic fluxes were involved in the synthesis of p-hydroxybenzoic acid (pHBA; the synthetic precursor of CoQ10), enhancing the anabolic flow of CoQ10. In addition, Ca2+ produced by the reaction of CaCO3 with organic acids promotes the synthesis of CoQ10. In summary, the use of CaCO3 adjustment is more favorable for the synthesis of CoQ10 by R. sphaeroides than NH3·H2O adjustment. The migration of metabolic flux caused by the perturbation of culture conditions was analyzed to compare the changes in the distribution of intracellular metabolic fluxes for the synthesis of CoQ10. Thus, the main nodes of the metabolic network were identified as G6P and chorismic acid. This provides a theoretical basis for the modification of genes related to the CoQ10 synthesis pathway.

The impact of utilizing oyster shell soil conditioner on the growth of tomato plants and the composition of inter-root soil bacterial communities in an acidic soil environment.

Introduction: The objective of this study is to examine the impact of various oyster shell soil conditioners, which are primarily composed of oyster shells, on the growth of tomatoes in acidic soil. Moreover, the aim of this investigation is to analyze the variety and structure of soil bacterial populations in close proximity to tomato roots while also contributing to the understanding of the physical, chemical, and biological mechanisms of oyster shell soil conditioners. Methods: Tomato plants were grown in acidic red soil in three groups: a control group and a treatment group that used two types of oyster shell soil conditioners, OS (oyster shell powder) and OSF (oyster shell powder with organic microbial fertilizer). A range of soil physicochemical properties were measured to study differences in inter-soil physicochemical parameters and the growth of tomato plantings. In addition, this study utilized the CTAB (Cetyltrimethylammonium Bromide) technique to extract DNA from the soil in order to investigate the effects of oyster shell soil conditioner on the composition and diversity of bacterial populations. Utilizing high-throughput sequencing technologies and diversity index analysis, the composition and diversity of bacterial populations in the soil adjacent to plant roots were then evaluated. Ultimately, correlation analysis was used in this study to explore the relationship between environmental factors and the relative abundance of soil bacteria in the inter-root zone of tomato plants. Results: The findings indicated that the oyster shell soil conditioners were capable of modifying the physicochemical characteristics of the soil. This was evidenced by significant increases in soil total nitrogen (16.2 and 59.9%), soil total carbon (25.8 and 27.7%), pH (56.9 and 55.8%), and electrical conductivity (377.5 and 311.7%) in the OS and OSF groups, respectively, compared to the control group (p < 0.05). Additionally, data pertaining to tomato seed germination and seedling growth biomass demonstrated that both oyster shell soil conditioners facilitated the germination of tomato seeds and the growth of seedlings in an acidic red clay soil (p < 0.05). On the other hand, the application of two oyster shell soil conditioners resulted in a modest reduction in the diversity of inter-root soil bacteria in tomato plants. Specifically, the group treated with OSF exhibited the most substantial fall in the diversity index, which was 13.6% lower compared to the control group. The investigation carried out on the soil between tomato plant roots yielded findings about the identification of the ten most abundant phyla. These phyla together represented 91.00-97.64% of the overall abundance. In the inter-root soil of tomatoes, a study identified four major phyla, namely Proteobacteria, Bacteroidetes, Acidobacteria, and Actinobacteria, which collectively accounted for up to 85% of the total abundance. At the general level, the relative abundance of Massilia increased by 2.18 and 7.93%, Brevundimonas by 5.43 and 3.01%, and Lysobacter by 3.12 and 7.49% in the OS and OSF groups, respectively, compared to the control group. However, the pathogenic bacteria unidentified_Burkholderiaceae decreased by 5.76 and 5.05%, respectively. The correlation analysis yielded conclusive evidence indicating that, which involved the use of CCA (Canonical Correlation Analysis) graphs and Spearman correlation coefficients, pH exhibited a positive correlation (p < 0.05) with Shewanella and a negative correlation (p < 0.05) with Bradyrhizobium. The relative abundance of Lysobacter and Massilia exhibited a positive correlation with the levels of total soil nitrogen. Discussion: The utilization of oyster shell soil conditioner on acidic red soil resulted in several positive effects. Firstly, it raised the pH level of the inter-root soil of tomato plants, which is typically acidic. This pH adjustment facilitated the germination of tomato seeds and promoted the growth of seedlings. In addition, the application of oyster shell soil conditioner resulted in changes in the structure of the bacterial community in the inter-root soil, leading to an increase in the relative abundance of Proteobacteria and Bacteroidetes and a decrease in the relative abundance of Acidobacteria. Furthermore, this treatment fostered the proliferation of genera of beneficial bacteria like Massilia, Brevundimonas, and Lysobacter, ultimately enhancing the fertility of the red soil.

A simple and rapid approach for on-site analysis of nicotine in tobacco based on a screen-printed electrode as an electrochemical sensor.

In this study, we report a portable kit consisting of a portable workstation, gold screen-printed electrode (SPE), 0.45 µm filter membrane, phosphate buffer solution (PBS), and acetic acid (1%) for point-of-use (POU) analysis of nicotine in tobacco. The activated-screen-printed electrode (A-SPE) displayed superior electron transmission efficiency, and the A-SPE without modification was employed for high-performance analysis of nicotine in actual tobacco after simple sample pretreatment. Remarkably, the fabricated nicotine sensor exhibited a broad working range of 10-100 µg g-1, a low limit of detection (LOD) of 6.4 µg mL-1, good stability, selectivity, and practicality under the optimal conditions. The method was applied to the determination of nicotine in (spiked) samples. Satisfactory recovery results demonstrated that the as-prepared portable kit method with outstanding electrocatalysis ability was feasible for analysis of nicotine in tobacco. Moreover, the values obtained using the A-SPE were in good agreement with those determined by gas chromatography-flame ionization detection (GC-FID), which confirms the feasibility and validity of the present method. The results of the as-proposed portable kit provided a new strategy for analyzing nicotine in actual tobacco samples.

Ultrasensitive monitoring of DNA damage associated with free radicals exposure using dynamic carbon nanotubes bridged interdigitated electrode array.

There are currently increasingly concerns over DNA damage related to free radicals due to their vital roles in human health, especially high-performance detection method. Herein, we report an ultra- sensitive monitoring of DNA damage associated with free radicals exposure using interdigitated electrode (IDE) array for the first time. The proposed IDE array was equipped with DNA-wrapped carbon nanotube-based bridges, which utilized the DNA damage mechanism due to the free radicals' attack and the efficient electrical detection nature of the interdigitated electrode. Experiments have been performed, and the results showed the device's capability for detecting DNA damage induced by multiple free radicals generated from different sources, including the Fenton reaction, UV radiation and cigarette smoke, showing the promising ability for DNA damage detection. In addition, the carbon nanotubes bridge-based interdigitated electrode sensor enabled different levels of sensing of DNA damage with great sensitivity and a wide detection range. It was illustrated that the ultrasensitive detection of free radicals generated from ultraviolet radiation (15 min - 125 min), cigarette smoke tar (1 µg/mL to 10 µg/mL) and Fenton reaction under different concentration of H2O2 (2.5 pM - 100 pM), have been detected successfully. Typically, the IDE array supports further performance improvement for the electrochemical detection in an ultrasensitive and high throughput route.

A facile approach for rapid on-site screening of nicotine in natural tobacco.

Nicotine (Nic) exposed to the environment which comes from tobacco products is the main addictive agent and specific classes of hazardous compound that merit concern. In this study, we have established a fast and reliable method to achieve specific detection of Nic in natural nicotiana tabacum within 30 s through a miniaturized platform based on screen printed gold electrode (SPE). A simple electrochemical pretreatment mean was employed on gold surface that led to the exposure of Au (111) facet and a convenient sample pretreatment method was adopted to realize the extraction of Nic in tobacco. The present electrochemical sensor exhibits an ample range of sensing from 10 µg/g to 200 µg/g, which is able to compliance with tobacco industry testing standards of actual samples. Over 60 sampling points from different origins in China or other countries were performed with direct analysis using this method and satisfactory results have been obtained. The proposed approach was demonstrated to be a very promising platform for significantly improving analytical efficiency in laboratories as well as for monitoring the source reduction control of Nic in the environment.

Novel sensing platform based on gold nanoparticle-aptamer and Fe-metal-organic framework for multiple antibiotic detection and signal amplification.

The development of a feasible antibiotic detection method is important in water quality analysis. In this study, we developed a metal-organic framework (MOF)-aptamer-3,3',5,5'-tetramethylbenzidine (TMB)-H2O2-based sensing platform composed of the reaction variable of TMB catalytic oxidation as the label (from colorless to blue) and aptamer as the target recognition element for antibiotic detection. The platform works by calculating the relation between the antibiotic concentration and the resultant decrease in MOF's catalytic activity. Basing from the comparison of typical iron-based MOF materials (Fe-MIL-53, Fe-MIL-88A, and Fe-MIL-100), we selected Fe-MIL-53 to obtain an improved signal amplification effect. The outstanding performance of the Fe-MIL-53-based sensing platform can be attributed to its topological flexibility and small electron transfer impedance. In addition, a signal increment of up to 86% was obtained with an intensified gold nanoparticle (AuNP)-supported aptamer. The inhibitory catalytic activity stemmed from the coating of antibiotic-(AuNP-aptamer) conjugates onto the outer surface of the MOF material, which increased the impedance and decreased the electron transfer efficiency. Validation results indicated that the platform showed high selectivity and sensitivity (i.e., wide linearity range of 50-200 nM, detection limit up to 8.1 ng/mL, and recovery rate of 106%-110%) for chloramphenicol detection and universal applicability for other antibiotics, including ampicillin, tetracycline, and oxytetracycline. In general, the detection reliability and easy operation of this platform render it a promising candidate for antibiotic detection in future water quality monitoring practices.

Graphene oxide wrapped melamine sponge as an efficient and recoverable adsorbent for Pb(II) removal from fly ash leachate.

Lead is one of the most toxic elements, which has been well recognized for its negative effect on the environment and human beings. But, preliminary methods such as chemical precipitation, membrane separation etc. and commonly used adsorbents based on adsorption technology were found to be expensive and inefficient. In this study, we modify the surface of melamine sponge (MS) with polydopamine (PDA) and then coat with glutathione/graphene oxide (GG) as the adsorbent (MS@GG) to removal Pb(II) from aqueous solutions and fly ash leachate. The maximum adsorption capacity of MS@GG was calculated to be 349.7 mg Pb/g GG, and the reaction reached equilibrium in 30 min which were both higher than raw GG material and most previously reported adsorbents due to active sites on the surface of GG, as well as the unique macroporous and hydrophilic structure of MS. Meanwhile, based on its easy separation, by using HCl as the regeneration agent, the materials revealed good reproducibility. In addition, when MS@GG was applied for the removal of Pb(II) in fly ash leachate, the removal efficiency reached up to 99.24%, indicating that the novel MS@GG was the promising candidate adsorbent material for Pb(II) removal.

N-Carbamoylmaleimide-treated carbon dots: stabilizing the electrochemical intermediate and extending it for the ultrasensitive detection of organophosphate pesticides.

To date, numerous methods have been reported for the detection of organophosphorus pesticides (OP) due to their severe potential hazard to the environment, public health and national security. However, very few works have ever found that the signal loss of thiocholine (TCh) during electrochemical processing is a key factor leading to the low sensitivity of acetylcholinesterase (AChE)-based OP electrochemical sensing platforms. Herein, we propose an ultrasensitive detection method for multiple OPs including parathion-methyl, paraoxon, dimethoate and O,O-dimethyl-O-2,2-dichlorovinyl-phosphate using N-carbamoylmaleimide-functionalized carbon dots (N-MAL-CDs) as a nano-stabilizer. For the first time, Michael addition is introduced into an AChE-based OP electrochemical sensing platform to enrich the electrochemical intermediate TCh. The Michael addition between TCh and N-MAL-CDs is demonstrated via XRD, FTIR, SEM and EDS elemental mapping experiments. Due to the stabilization and enhancement of TCh with N-MAL-CDs, the as prepared OP sensing platform achieves ultrahigh sensitivity by detecting the initial electrochemical signals of TCh without signal loss, showing a wide linear range of 3.8 × 10-15-3.8 × 10-10 M for parathion-methyl and 1.8 × 10-14-3.6 × 10-10 M for paraoxon, with a limit of detection of 1.4 × 10-15 M for parathion-methyl and 4.8 × 10-15 M for paraoxon.