Mesoporous carbon decorated with MIL-100(Fe) as an electrochemical platform for ultrasensitive determination of trace cadmium and lead ions in surface water.

In this work, MIL-100(Fe)-decorated mesoporous carbon powders (MC@MIL-100(Fe)) were prepared by in situ growth of MIL-100(Fe) on the surface of ZIF-8 framework-based mesoporous carbons (MC). The hybrid material was characterized using SEM equipped with EDS mapping for morphology investigation, X-ray photoelectron spectroscopy for chemical valence analysis, and X-ray diffraction for crystal structure determination. The developed sensor separated from the traditional bismuth film decoration, and simultaneously, MC@MIL-100(Fe) was applied for the first time to electrochemically detect trace amounts of Pb(II) and Cd(II). The fabricated MC@MIL-100(Fe)-based electrochemical sensor showed excellent response to the target analytes at -0.55 and - 0.75 V for lead and cadmium ions, respectively. By adjusting some measurement parameters, that is, the loading concentration of MC@MIL-100(Fe), acidity of the HAc-NaAc buffer (ABS), deposition potential, and deposition time, the analytical performance of the proposed electrochemical sensor was examined by exploring the calibration curve, repeatability, reproducibility, stability, and anti-interference under optimized conditions. The response current of the proposed MC@MIL-100(Fe) electrochemical sensor showed a well-defined linear relationship in the concentration ranges of 2-250 and 2-270 µg·L-1 for Cd(II) and Pb(II), respectively. In addition, the detection limits of the sensor for Cd(II) and Pb(II) were 0.18 and 0.15 µg L-1, respectively, which are well below the World Health Organization (WHO) drinking water guideline value. The MC@MIL-100(Fe) can be potentially used as an electrochemical platform for monitoring heavy metals in surface water, with satisfactory results.

Two-dimensional BCN nanosheets self-assembled with hematite nanocrystals for sensitively detecting trace toxic Pb(II) ions in natural water.

In the present work, hematite-boron-carbonitride (Fe2O3-BCN) nanosheets were synthesized by a simple hydrothermal reaction and the following high temperature treatment. The morphology, structure and chemical composition of the as-prepared material were carefully characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The Fe2O3-BCN nanosheets were used to modified on the surface of the glassy carbon electrode to fabricate an electrochemical sensor for lead ions (Pb(II)) via differential pulse anodic stripping voltammetry (DPASV). At the same time, the influence of the modification concentration, solution acidity, deposition potential and deposition time on response peak current of Pb(II) at the Fe2O3-BCN-based electrochemical sensor was well investigated. Under the optimized conditions, the electrochemical signal and concentration of Pb(II) show two-stage linear relationship in the range of 0.5 - 40 µg/L and 40 -140 µg/L, with a limit of detection (LOD) of 0.129 µg/L. The Fe2O3-BCN-based electrochemical sensor shows excellent selectivity and anti-interference ability in the anti-interference experiments and actual sample analysis experiments, revealing its broad application in environmental monitoring of Pb(II).

Release of glucose repression on xylose utilization in Kluyveromyces marxianus to enhance glucose-xylose co-utilization and xylitol production from corncob hydrolysate.

BACKGROUND: Lignocellulosic biomass is one of the most abundant materials for biochemicals production. However, efficient co-utilization of glucose and xylose from the lignocellulosic biomass is a challenge due to the glucose repression in microorganisms. Kluyveromyces marxianus is a thermotolerant and efficient xylose-utilizing yeast. To realize the glucose-xylose co-utilization, analyzing the glucose repression of xylose utilization in K. marxianus is necessary. In addition, a glucose-xylose co-utilization platform strain will facilitate the construction of lignocellulosic biomass-utilizing strains. RESULTS: Through gene disruption, hexokinase 1 (KmHXK1) and sucrose non-fermenting 1 (KmSNF1) were proved to be involved in the glucose repression of xylose utilization while disruption of the downstream genes of cyclic AMP-protein kinase A (cAMP-PKA) signaling pathway or sucrose non-fermenting 3 (SNF3) glucose-sensing pathway did not alleviate the repression. Furthermore, disruption of the gene of multicopy inhibitor of GAL gene expression (KmMIG1) alleviated the glucose repression on some nonglucose sugars (galactose, sucrose, and raffinose) but still kept glucose repression of xylose utilization. Real-time PCR analysis of the xylose utilization related genes transcription confirmed these results, and besides, revealed that xylitol dehydrogenase gene (KmXYL2) was the critical gene for xylose utilization and stringently regulated by glucose repression. Many other genes of candidate targets interacting with SNF1 were also evaluated by disruption, but none proved to be the key regulator in the pathway of the glucose repression on xylose utilization. Therefore, there may exist other signaling pathway(s) for glucose repression on xylose consumption. Based on these results, a thermotolerant xylose-glucose co-consumption platform strain of K. marxianus was constructed. Then, exogenous xylose reductase and xylose-specific transporter genes were overexpressed in the platform strain to obtain YHY013. The YHY013 could efficiently co-utilized the glucose and xylose from corncob hydrolysate or xylose mother liquor for xylitol production (> 100 g/L) even with inexpensive organic nitrogen sources. CONCLUSIONS: The analysis of the glucose repression in K. marxianus laid the foundation for construction of the glucose-xylose co-utilizing platform strain. The efficient xylitol production strain further verified the potential of the platform strain in exploitation of lignocellulosic biomass.

Metabolic engineering of indole pyruvic acid biosynthesis in Escherichia coli with tdiD.

BACKGROUND: Indole pyruvic acid (IPA) is a versatile platform intermediate and building block for a number of high-value products in the pharmaceutical and food industries. It also has a wide range of applications, such as drugs for the nervous system, cosmetics, and luminophores. Chemical synthesis of IPA is a complicated and costly process. Moreover, through the biosynthesis route employing L-amino acid oxidase, the byproduct hydrogen peroxide leads the degradation of IPA. TdiD, identified as a specific tryptophan aminotransferase, could be an alternative solution for efficient IPA biosynthesis. RESULTS: Escherichia coli strain W3110, which demonstrates basic production when supplied with tryptophan, was engineered for IPA biosynthesis. Several strategies were implemented to improve IPA production. First, through incorporating the codon-optimized tdiD into W3110, IPA levels increased from 41.54 ± 1.26 to 52.54 ± 2.08 mg/L. Second, after verifying the benefit of an increased phenylpyruvate pool, a YL03 strain was constructed based on a previously reported mutant strain of W3110 with a plasmid carrying aroF fbr and pheA fbr to further improve IPA production. The recombinant YL03 strain accumulated IPA at 158.85 ± 5.36 mg/L, which was 3.82-fold higher than that of the wild-type W3110 strain. Third, optimization of tdiD co expression was carried out by replacing the Trc promoter with a series of constitutively active promoters along with increasing the plasmid copy numbers. The highest IPA production was observed in YL08, which achieved 236.42 ± 17.66 mg/L and represented a greater than 5-fold increase as compared to W3110. Finally, the effects of deletion and overexpression of tnaA on IPA biosynthesis were evaluated. The removal of tnaA led to slightly reduced IPA levels, whereas the overexpression of tnaA resulted in a considerable decline in production. CONCLUSIONS: This study illustrates the feasibility of IPA biosynthesis in E. coli through tdiD. An efficient IPA producing strain, YL08, was developed, which provides a new possibility for biosynthesis of IPA. Although the final production was limited, this study demonstrates a convenient method of IPA synthesis.

Discovery of Metal Ions Chelator Quercetin Derivatives with Potent Anti-HCV Activities.

Analogues or isosteres of α,γ-diketoacid (DKA) 1a show potent inhibition of hepatitis C virus (HCV) NS5B polymerase through chelation of the two magnesium ions at the active site. The anti-HCV activity of the flavonoid quercetin (2) could partly be attributed to it being a structural mimic of DKAs. In order to delineate the structural features required for the inhibitory effect and improve the anti-HCV potency, two novel types of quercetin analogues, 7-O-arylmethylquercetins and quercetin-3-O-benzoic acid esters, were designed, synthesized and evaluated for their anti-HCV properties in cell-based assays. Among the 38 newly synthesized compounds, 7-O-substituted derivative 3i and 3-O-substituted derivative 4f were found to be the most active in the corresponding series (EC50 = 3.8 µM and 9.0 µΜ, respectively). Docking studies suggested that the quercetin analogues are capable of establishing key coordination with the two magnesium ions as well as interactions with residues at the active site of HCV NS5B.

Measurement and spatial correlation analysis of high-quality development Level: A case study of the Yangtze River Delta urban agglomeration in China.

Against the backdrop of slowing economic growth and increasing environmental pressure, the Yangtze River Delta city cluster, as one of the largest city clusters in the world, has become more driven in its pursuit of high-quality development. We constructed a system of 24 evaluation indexes and used entropy-weighted TOPSIS to calculate and study the high-quality development index of urban agglomerations in the region. First, the level of high quality development (HQD) of the Yangtze River Delta city cluster generally improved from 2010 to 2021, with 2017 was the best year, while 2010 was the worst year. Second, in the multidimensional evaluation of HQD, Jiangsu excels in innovation and people's livelihood with 0.524 and 0.534, respectively; Shanghai (0.531) excels in coordinated development; Zhejiang excels in green and economic development with 0.557 and 0.484, respectively; and Anhui lags behind in all aspects. Third, the development process of HQD in the Yangtze River Delta region is uneven, and the level of HQD development varies greatly among the city clusters in the province. The measurement results show that Shanghai (0.511) has the highest score, followed by Zhejiang (0.484), Jiangsu (0.440) and Anhui (0.435). Fourth, spatial correlation analysis shows that Shanghai and Jiangsu are mainly distributed in the double-high region, Zhejiang is distributed in the high-low region, while Anhui is concentrated in the low-low region. The results of this study help us understand more deeply the characteristics and challenges of high-quality development in the Yangtze River Delta urban agglomerations and provide a scientific basis for more precise urban development policies.

Research on tourism ecological safety evaluation of Huizhou Cultural and ecological reserve based on entropy -TOPSIS.

Tourism ecological security is the basic guarantee for the sustainable development of tourist sites, Huizhou Cultural and Ecological Reserve is an important area for the im-plementation of ecological protection in China, and it is of great significance to carry out research on tourism ecological security. The study adopted the DPSIR model to construct a comprehensive evaluation index system for tourism ecological security and used entropy value-TOPSIS and ArcGIS software to analyze the inter-annual changes and spatial change characteristics of tourism ecological security in the study area. The results show that: firstly, the comprehensive index of tourism ecological safety in the study area from 2010 to 2021 shows a trend of "decreasing-increasing" and an overall increasing trend; secondly, all the sub-systems show an increasing or stabilizing state in recent years during 2010-2021; the state and response sub-systems show an increas-ing or stabilizing state in recent years; and the state and response sub-systems show an increasing trend in recent years. Secondly, all the subsystems showed an increase or stabilization in recent years between 2010 and 2021, and the state and response sub-systems are the main systems to improve the ecological safety of tourism in the study area; thirdly, the difference in the level of ecological safety of tourism in each county of the study area increased and then narrowed from 2010 to 2021, and the change of safety level usually shifted between neighboring levels, and the probability of transfer-ring across the levels was relatively small. , Shexian County, Yixian County, Qimen County, Tunxi District, and the tourism eco-safety level of Huangshan District, Hui-zhou District, Jixi County, and Xiuning County increased at a faster rate than other counties. The study further extends the scale to the district and county level, tries to explore the relevant factors affecting the ecological security of tourism, and proposes countermeasures for the sustainable development of the study area based on the re-sults, which will bring some reference value to the ecological governance and policy formulation of this kind of research.

Tetrazole and triazole as bioisosteres of carboxylic acid: discovery of diketo tetrazoles and diketo triazoles as anti-HCV agents.

A series of diketo tetrazoles and diketo triazoles were designed and synthesized as bioisosteres of α,γ-diketo acid, the active site inhibitor of HCV (Hepatitis C virus) polymerase NS5B. Among the synthesized compounds, 4-(4-fluorobenzyloxy)phenyl diketo triazole (30) exhibited anti-HCV activity with an EC50 value of 3.9 µM and an SI value more than 128. The reduction of viral protein and mRNA levels were also validated, supporting the anti-HCV activity of compound 30. These results provide convincing evidence that the diketo tetrazoles and diketo triazoles can be developed as bioisosteres of α,γ-diketo acid to exhibit potent inhibitory activity against HCV.

Strong water-resistant, UV-blocking cellulose/glucomannan/lignin composite films inspired by natural LCC bonds.

Lignocellulose multi-component films were fabricated using cellulose nanofiber (CNF), konjac glucomannan (KGM), and alkali lignin (AL). Covalent bonds among these three lignocellulose components were constructed with epichlorohydrin (EPI) to achieve chemical cross-linking, which inspired by lignin-carbohydrate complex (LCC) bonds in natural wood. Chemical cross-linking enhanced the mechanical properties of the composite films, the tensile stress reached to 110.47 MPa and the tensile strain reached to 8.27%. The presence of hemicellulose and lignin in composite films contributed to the functionalities of the films. The composite film with 20 wt% AL blocked over 99% of both UVA and UVB, the water contact angle reached 110.38°, and no obvious swelling behavior was observed after soaking in water for 180 days. Composite films with strong water-resistance and UV-blocking have great potential for the related application.

Effects of Carbonaceous Materials with Different Structures on Cadmium Fractions and Microecology in Cadmium-Contaminated Soils.

Carbonaceous materials have proved to be effective in cadmium remediation, but their influences on soil microecology have not been studied well. Taking the structural differences and the maintenance of soil health as the entry point, we chose graphene (G), multi-walled carbon nanotubes (MWCNTs), and wetland plant-based biochar (ZBC) as natural and engineered carbonaceous materials to explore their effects on Cd fractions, nutrients, enzyme activities, and microbial communities in soils. The results showed that ZBC had stronger electronegativity and more oxygen-containing functional groups, which were related to its better performance in reducing soil acid-extractable cadmium (EX-Cd) among the three materials, with a reduction rate of 2.83-9.44%. Additionally, ZBC had greater positive effects in terms of improving soil properties, nutrients, and enzyme activities. Redundancy analysis and correlation analysis showed that ZBC could increase the content of organic matter and available potassium, enhance the activity of urease and sucrase, and regulate individual bacterial abundance, thereby reducing soil EX-Cd. Three carbonaceous materials could maintain the diversity of soil microorganisms and the stability of the microbial community structures to a certain extent, except for the high-dose application of ZBC. In conclusion, ZBC could better immobilize Cd and maintain soil health in a short period of time.

Impacts of exogenous mineral silicon on cadmium migration and transformation in the soil-rice system and on soil health.

Exogenous silicon has been shown to enhance plant growth and alleviate heavy metals toxicity, but the regulation mechanism of silicon on cadmium migration and transformation in the soil-rice system is still unclear, which is worth further study. In this study, a pot experiment was carried out to explore the influence of different doses (0, 1 and 5%) of mineral silicon on soil properties, nutrient availability, rice growth, soil enzyme activities, Cd bioavailability, and uptake and accumulation of Cd in high-accumulating (H) and low-accumulating (L) rice cultivars grown in contaminated soils. Results showed that mineral-Si treatment could increase the total biomass and grain yield, with an increased rate of 17.7-27.3% and 14.7-19.1% for H; while 26.2-33.4% and 21.3-30.3% for L. Compared with non-mineral-Si treatment, the soil EX-Cd decreased by 3.9-13.3% (H) and 2.3-10.7% (L). Additionally, the Cd content in rice grain was significantly declined by 29.5-31.3% (H) and 34.9-35.2% (L). Mineral-Si enhanced urease, sucrase, and neutral phosphatase activities in both cultivars, but suppressed catalase activity in H. A selective change in bacterial community structure was observed under mineral-Si treatment, however, the bacterial community remained stable, suggesting that the mineral-Si had no adverse effect on the microbial community. The positive response of soil enzymes activities, rice growth and the overall stabilization of microbial environment for mineral-Si addition to the Cd contaminated soils indicated that mineral-Si could mitigate the risk of Cd and well maintain the soil health, proving it to be eco-friendly and low-cost amendment for soils remediation.

Efficient l-lactic acid production from corncob residue using metabolically engineered thermo-tolerant yeast.

Lactic acid is an important industrial product and the production from inexpensive and renewable lignocellulose can reduce the cost and environmental pollution. In this study, a Kluyveromyces marxianus strain which produced lactic acid efficiently from corncob was constructed. Firstly, two of six different lactate dehydrogenases, which from Plasmodium falciparum and Bacillus subtilis, respectively, were proved to be effective for l-lactic acid production. Then, five single genetic modifications were conducted. The overexpression of Saccharomyces cerevisiae proton-coupled monocarboxylate transporter, K. marxianus 6-phosphofructokinase, or disruption of K. marxianus putative d-lactate dehydrogenase enhanced the l-lactic acid accumulation. Finally, the strain YKX071, obtained via combination of above effective genetic engineering, produced 103.00 g/L l-lactic acid at 42 °C with optical purity of 99.5% from corncob residue via simultaneous saccharification and co-fermentation. This study first developed an effective platform for high optical purity l-lactic acid production from lignocellulose using yeast with inexpensive nitrogen sources.

Engineered Kluyveromyces marxianus for pyruvate production at elevated temperature with simultaneous consumption of xylose and glucose.

Xylose and glucose from lignocellulose are sustainable sources for production of pyruvate, which is the starting material for the synthesis of many drugs and agrochemicals. In this study, the pyruvate decarboxylase gene (KmPDC1) and glycerol-3-phosphate dehydrogenase gene (KmGPD1) of Kluyveromyces marxianus YZJ051 were disrupted to prevent ethanol and glycerol accumulation. The deficient growth of PDC disruption was rescued by overexpressing mutant KmMTH1-ΔT. Then pentose phosphate pathway and xylitol dehydrogenase SsXYL2-ARS genes were overexpressed to obtain strain YZB053 which produced pyruvate with xylose other than glucose. It produced 24.62g/L pyruvate from 80g/L xylose with productivity of 0.51g/L/h at 42°C. Then, xylose-specific transporter ScGAL2-N376F was overexpressed to obtain strain YZB058, which simultaneously consumed 40g/L glucose and 20g/L xylose and produced 29.21g/L pyruvate with productivity of 0.81g/L/h at 42°C. Therefore, a platform for pyruvate production from glucose and xylose at elevated temperature was developed.

Directed evolution to improve the catalytic efficiency of urate oxidase from Bacillus subtilis.

Urate oxidase is a key enzyme in purine metabolism and catalyzes the oxidation of uric acid to allantoin. It is used to treat hyperuricemia and gout, and also in a diagnostic kit. In this study, error-prone polymerase chain reaction and staggered extension process was used to generate a mutant urate oxidase with improved enzyme activity from Bacillus subtilis. After several rounds of mutagenesis and screening, two mutants 6E9 and 8E279 were obtained which exhibited 2.99 and 3.43 times higher catalytic efficiency, respectively. They also exhibited lower optimal reaction temperature and higher thermo-stability. D44V, Q268R and K285Q were identified as the three most beneficial amino acid substitutions introduced by site-directed mutagenesis. D44V/Q268R, which was obtained through random combination of the three mutants, displayed the highest catalytic activity. The Km, kcat/Km and enzyme activity of D44V/Q268R increased by 68%, 83% and 129% respectively, compared with that of wild-type urate oxidase. Structural modeling indicated that mutations far from the active site can have significant effects on activity. For many of them, the underlying mechanisms are still difficult to explain from the static structural model. We also compared the effects of the same set of single point mutations on the wild type and on the final mutant. The results indicate strong effects of epistasis, which may imply that the mutations affect catalysis through influences on protein dynamics besides equilibrium structures.