Microwave-assisted and methanol/acetic acid-free method for rapid staining of proteins in SDS-PAGE gels.

We describe a microwave-assisted, methanol and acetic acid-free, inexpensive method for rapid staining of SDS-PAGE proteins. Only citric acid, benzoic acid, and Coomassie brilliant blue G-250 (CBG) were used. Microwave irradiation reduced the detection duration, and proteins in a clear background were visualized within 30 min of destaining, after 2 min of fixing and 12 min of staining. By using this protocol, comparable band intensities were obtained to the conventional methanol/acetic acid method.

Smartphone-based dual inverse signal MOFs fluorescence sensing for intelligent on-site visual detection of malachite green.

The development of intelligent, sensitive, and visual methods for the rapid detection of veterinary drug residues is essential to ensure food quality and safety. Here, a smartphone-based dual inverse signal MOFs fluorescence sensing system was proposed for intelligent in-site visual detection of malachite green (MG). A UiO-66-NH2@RhB-dual-emission fluorescent probe was successfully synthesized in one step using a simple one-pot method. The inner filter effect (IFE) quenches the red fluorescence, while hydrogen bonding interaction enhances the blue fluorescence, enabling highly sensitive, accurate, and visual detection of MG dual inverse signals through fluorescence analysis. The probe showed great linearity over a wide range of 0.1-100 µmol/L, with a limit of detection (LOD) of 20 nmol/L. By integrating smartphone photography and RGB (red, green, and blue) analysis, accurate quantitative analysis of MG in water and actual fish samples can be achieved within 5 min. This developed platform holds great promise for the on-site detection of MG in practical applications, with the advantages of simplicity, cost-effectiveness, and rapidity. Consequently, it may open up a new pathway for on-site evaluation of food safety and environmental health.

Microalgae derived honeycomb structured mesoporous diatom biosilica for adsorption of malachite green: Process optimization and modeling.

The present study investigated the removal of malachite green dye from aquifers by means of microalgae-derived mesoporous diatom biosilica. The various process variables (dye concentration, pH, and adsorbent dose) influencing the removal of the dye were optimized and their interactive effects on the removal efficiency were explored by response surface methodology. The pH of the solution (pH = 5.26) was found to be the most dominating among other tested variables. The Langmuir isotherm (R2 = 0.995) best fitted the equilibrium adsorption data with an adsorption capacity of 40.7 mg/g at 323 K and pseudo-second-order model (R2 = 0.983) best elucidated the rate of dye removal (10.6 mg/g). The underlying mechanism of adsorption was investigated by Weber-Morris and Boyd models and results revealed that the film diffusion governed the overall adsorption process. The theoretical investigations on the dye structure using DFT-based chemical reactivity descriptors indicated that malachite green cations are electrophilic, reactive and possess the ability to accept electrons, and are strongly adsorbed on the surface of diatom biosilica. Also, the Fukui function analysis proposed the favorable adsorption sites available on the adsorbent surface.

Tecoma stans floral extract-mediated synthesis of MgFe2O4/ZnO nanoparticles for adsorption and photocatalytic degradation of coomassie brilliant blue dye.

Toxic organic dyes-containing wastewater treatment by adsorption and photocatalytic techniques is widely applied, but adsorbents and photocatalysts are often synthesized through chemical methods, leading to secondary pollution by released chemicals. Here, we report a benign method using Tecoma stans floral extract to produce MgFe2O4/ZnO (MGFOZ) nanoparticles for adsorption and photocatalytic degradation of coomassie brilliant blue (CBB) dye. Green MGFOZ owned a surface area of 9.65 m2/g and an average grain size of 54 nm. This bio-based nanomaterial showed higher removal percentage and better recyclability (up to five cycles) than green MgFe2O4 and ZnO nanoparticles. CBB adsorption by MGFOZ was examined by kinetic and isotherm models with better fittings of Bangham and Langmuir or Temkin. RSM-based optimization was conducted to reach an actual adsorption capacity of 147.68 mg/g. Moreover, MGFOZ/visible light system showed a degradation efficiency of 89% CBB dye after 120 min. CBB adsorption can be controlled by both physisorption and chemisorption while •O2- and •OH radicals are responsible for photo-degradation of CBB dye. This study suggested that MGFOZ can be a promising adsorbent and catalyst for removal of organic dyes in water.

Symmetry-breaking malachite green as a near-infrared light-activated fluorogenic photosensitizer for RNA proximity labeling.

Cellular RNA is asymmetrically distributed in cells and the regulation of RNA localization is crucial for proper cellular functions. However, limited chemical tools are available to capture dynamic RNA localization in complex biological systems with high spatiotemporal resolution. Here, we developed a new method for RNA proximity labeling activated by near-infrared (NIR) light, which holds the potential for deep penetration. Our method, termed FAP-seq, utilizes a genetically encoded fluorogen activating protein (FAP) that selectively binds to a set of substrates known as malachite green (MG). FAP binding restricts the rotation of MG and rapidly activates its fluorescence in a wash-free manner. By introducing a monoiodo modification to MG, we created a photosensitizer (MG-HI) with the highest singlet oxygen generation ability among various MG derivatives, enabling both protein and RNA proximity labeling in live cells. New insights are provided in the transcriptome analysis with FAP-seq, while a deeper understanding of the symmetry-breaking structural arrangement of FAP-MG-HI was obtained through molecular dynamics simulations. Overall, our wash-free and NIR light-inducible RNA proximity labeling method (FAP-seq) offers a powerful and versatile approach for investigating complex mechanisms underlying RNA-related biological processes.

Removal of malachite green from water: Comparison of adsorption in a residue-derived AC versus photocatalytic oxidation with TiO2 and study of the adsorption-photocatalysis synergy.

The literature rarely compiles studies devoted to the removal of pollutants in aqueous media comparing adsorption and photocatalytic degradation, and does not pay enough attention to the analysis of combined adsorption-photocatalytic oxidation processes. In the present manuscript, the removal of malachite green (MG) from aqueous solutions has been investigated in three different sustainable scenarios: i) adsorption on activated carbon (AC) derived from a residue, luffa cylindrica, ii) photocatalytic oxidation under simulated solar light using titanium dioxide (TP) and iii) combined adsorption-photocatalytic oxidation using TP-AC (70/30 wt./wt.) under simulated solar light. The study has revealed that in the three scenarios and studied conditions, the total removal of this endocrine-disrupting dye from the solution takes place in the assayed time, 2 h, in some cases just in a few minutes. MG adsorption in the AC is a very fast and efficient removal method. MG photocatalytic oxidation with TP also occurs efficiently, although the oxidized MG is not totally mineralized. MG removal using the TP-AC composite under simulated solar light occurs only slightly faster to the MG adsorption in the AC, being adsorption the dominating MG removal mechanism for TP-AC. Thus, more than 90% of the removed MG with TP-AC under simulated solar light is adsorbed in this carbon-containing composite. The obtained results highlight the interest in adsorption, being the selection of the most suitable removal method dependent on several factors (i.e., the cost of the AC regeneration, for adsorption, or the toxicity of the intermediate oxidation species, for photooxidation). Paying attention to MG photooxidation with TiO2, comparison of two working photodegradation schemes shows that the direct photodegradation of MG from solution, avoiding any initial dark equilibrium period, is more efficient from a time perspective. The use of scavengers has proved that MG photodegradation occurs via an oxidation mechanism dominated by superoxide anion radicals.

Co-immobilizing laccase-mediator system by in-situ synthesis of MOF in PVA hydrogels for enhanced laccase stability and dye decolorization efficiency.

The laccase mediator system (LMS) with a broad substrate range has attracted much attention as an efficient approach for water remediation. However, the practical application of LMS is limited due to their high solubility, poor stability and low reusability. Herein, the bimetallic Cu/ZIFs encapsulated laccase was in-situ grown in poly(vinyl alcohol) (PVA) polymer matrix. The PVA-Lac@Cu/ZIFs hydrogel was formed via one freeze-thawing cycle, and its catalytic stability was significantly improved. The mediator was further co-immobilized on the hydrogel, and this hierarchically co-immobilized ABTS/PVA-Lac@Cu/ZIFs hydrogel could avoid the continuous oxidation reaction between laccase and redox mediators. The co-immobilized LMS biocatalyst was used to degrade malachite green (MG), and the degradation rate was up to 100 % within 4 h. More importantly, the LMS could be recycled synchronously from the dye solutions and reused to degrade MG multiple times. The degradation rate remained above 69.4 % after five cycles. Furthermore, the intermediate products were detected via liquid chromatography-mass spectrometry, and the potential degradation pathways were proposed. This study demonstrated the significant potential of utilizing the MOF nanocrystals and hydrogel as a carrier for co-immobilized LMS, and the effective reuse of both laccase and mediator was promising for laccase application in wastewater treatment.

Effective removal of malachite green oxalate from aqueous solution using Newbouldia laevis husk/MWCNTs nanocomposite: equilibrium, kinetics, and thermodynamics.

The discharge of colored effluent into water bodies is a big concern; hence, the current work was designed to fabricate a superior nanocomposite (NBM) using the Newbouldia laevis husk (NB) and functionalized multiwalled carbon nanotubes (f-MWCNTs) for the adsorption of malachite green oxalate (MGO). Brunauer-Emmett-Teller (BET) surface analysis was used to assess the specific surface area of NB (0.7699 m2 g-1) and NBM (94.006 m2 g-1). Fourier transform infrared spectroscopy (FTIR) was employed to determine the chemical moieties on the surface of the adsorbent. Field emission scanning electron microscopy (FESEM) and thermogravimetric analysis (TGA) were used to analyze the surface morphology and the thermal behavior of the adsorbents. Essential factors of the adsorption process were investigated, and it was revealed that pH 6.0, adsorbent dose of 0.05 g, contact time 80 min, concentration of 100 mg dm-3 and maximum adsorption capacity of 35.78 mg g-1 (NB) and 69.97 mg g-1 (NBM) were the optimal parameters. The NB and NBM adsorption processes followed a pseudo-first-order kinetic model. The exothermic and endothermic adsorptive processes were noticed to be the best descriptions of MGO elimination by NB and NBM, respectively. The uptake of MGO by NB and NBM was best described by models of Freundlich and Langmuir isotherms. Besides, NBM demonstrated uptake efficiency that is >80% after the fourth adsorption/desorption cycle. As a result, NBM has a wide range of possible uses in environmental remediation.

The husk of Newbouldia laevis is a frequent waste that must be managed properly. This paper describes the application of Newbouldia laevis husk as a value-added material for the design of a water treatment agent. The use of carbon nanotube in the modification of Newbouldia laevis husk would have a synergistic effect on the overall property of the nanocomposite. Nanocomposite synthesized from multiwalled carbon nanotubes (MWCNTs) and Newbouldia laevis husk were characterized and used for the sequestration of malachite green oxalate from contaminated water. Our primary goal is to optimize the nanocomposite by varying factors of adsorption such as solution pH, equilibrium, kinetic, thermodynamic, and regeneration studies. We believe that this study will contribute to the existing knowledge of Newbouldia laevis husk. Owing to the exceptional potential of the nanocomposite, this adsorbent can be extended to possible field applications.

Efficient and Selective Adsorption of Cationic Dye Malachite Green by Kiwi-Peel-Based Biosorbents.

In this study, pristine kiwi peel (KP) and nitric acid modified kiwi peel (NA-KP) based adsorbents were prepared and evaluated for selective removal of cationic dye. The morphology and chemical structure of KP and NA-KP were fully characterized and compared, and results showed nitric acid modification introduced more functional groups. Moreover, the adsorption kinetics and isotherms of malachite green (MG) by KP and NA-KP were investigated and discussed. The results showed that the adsorption process of MG onto KP followed a pseudo-second-order kinetic model and the Langmuir isotherm model, while the adsorption process of MG onto NA-KP followed a pseudo-first-order kinetic model and the Freundlich isotherm model. Notably, the Langmuir maximum adsorption capacity of NA-KP was 580.61 mg g-1, which was superior to that of KP (297.15 mg g-1). Furthermore, thermodynamic studies demonstrated the feasible, spontaneous, and endothermic nature of the adsorption process of MG by NA-KP. Importantly, NA-KP showed superior selectivity to KP towards cationic dye MG against anionic dye methyl orange (MO). When the molar ratio of MG/MO was 1:1, the separation factor (αMG/MO) of NA-KP was 698.10, which was 5.93 times of KP. In addition, hydrogen bonding, π-π interactions, and electrostatic interaction played important roles during the MG adsorption process by NA-KP. This work provided a low-cost, eco-friendly, and efficient option for the selective removal of cationic dye from dyeing wastewater.

Removal of the Basic and Diazo Dyes from Aqueous Solution by the Frustules of Halamphora cf. salinicola (Bacillariophyta).

Industrial wastes with hazardous dyes serve as a major source of water pollution, which is considered to have an enormous impact on public health. In this study, an eco-friendly adsorbent, the porous siliceous frustules extracted from the diatom species Halamphora cf. salinicola, grown under laboratory conditions, has been identified. The porous architecture and negative surface charge under a pH of 7, provided by the various functional groups via Si-O, N-H, and O-H on these surfaces, revealed by SEM, the N2 adsorption/desorption isotherm, Zeta-potential measurement, and ATR-FTIR, respectively, made the frustules an efficient mean of removal of the diazo and basic dyes from the aqueous solutions, 74.9%, 94.02%, and 99.81% against Congo Red (CR), Crystal Violet (CV), and Malachite Green (MG), respectively. The maximum adsorption capacities were calculated from isotherms, as follows: 13.04 mg g-1, 41.97 mg g-1, and 33.19 mg g-1 against CR, CV, and MG, respectively. Kinetic and isotherm models showed a higher correlation to Pore diffusion and Sips models for CR, and Pseudo-Second Order and Freundlich models for CV and MG. Therefore, the cleaned frustules of the thermal spring-originated diatom strain Halamphora cf. salinicola could be used as a novel adsorbent of a biological origin against anionic and basic dyes.

Dye degradation by early colonizing marine bacteria from the Arabian Sea, India.

Malachite green dye belongs to the triphenylmethane group and is a common environmental pollutant that threatens non-target organisms. We report the potential of the early colonizing marine bacterium Pseudomonas sp. ESPS40 isolated from the Arabian Sea, India, to decolorize malachite green (MG). The bacterium ESPS40 exhibited a higher ability for MG degradation (86-88%) at varying NaCl concentrations (1-3%). The highest MG degradation (~ 88%) was observed at 1% NaCl. The bacterial strain ESPS40 showed degradation up to 800 mg L-1 MG. Further, enzyme activities such as tyrosinase (63.48-526.52 U L-1) and laccase (3.62-28.20 U L-1) were also analyzed with varying concentrations (100 mg L-1-1000 mg L-1) of MG during the degradation process. The dye degradation was confirmed by Fourier transform infrared spectroscopy (FTIR) and high-performance liquid chromatography (HPLC). The outcome of the present study demonstrated Pseudomonas sp. ESPS40 as a potential strain for the efficient degradation of MG at higher concentrations. Thus, Pseudomonas sp. ESPS40 can be utilized as a potential candidate for the biodegradation of MG in wastewater treatment.

Malachite green and leucomalachite green in fish: a global systematic review and meta-analysis.

Malachite green (MG), an antibiotic with antifungal activity, is illegally used in aquaculture. Given that this chemical is teratogenic and mutagenic, abstinence from intake seems to be a need for public safety. The goal of this systematic review and meta-analysis was to determine the global contamination of fishes by MG and its reduced metabolite, leucomalachite green (LMG), in a number of marine and farmed fish species. For literature published prior to January 2022, several databases (Web of Science, PubMed, and Scopus) were investigated. In total, 20 publications (10 countries, 724 samples) achieved the criteria for inclusion. The overall average MG and LMG concentrations were 0.48 (95% CI: 0.47, 0.49 µg kg-1) and 0.59 (95% CI: 0.39, 0.79 µg/kg-1), respectively. Eel (M. albus) 15.50 (95% CI: (14.39, 45.39 µg kg-1) and eel (A. anguilla) 4.46 (95% CI: 1.23, 7.69 µg kg-1) had the greatest contamination of MG and LMG, according to the effect size, respectively. Warm-water fish had a concentration of 2.591 (95% CI: 2.25, 2.93 µg kg-1) while cold-water fish had a concentration of 1.55 (95% CI: 0.25, 2.84 µg kg-1). Fish containing medium-fat level of 1.86 (95% CI: 1.27, 2.44 µg kg-1) and high-fat content of 1.10 (95% CI: 0.93, 1.26 µg kg-1) had accumulate MG and LMG in their tissues, respectively. As a result, total MG observed in countries including China, Iran, and the Netherlands was higher than authorized (2 µg kg-1). The toxicity of MG and LMG demands more monitoring, especially in countries where these chemicals' residues are significant.

Removal of dyes (BG, MG, and SA) from aqueous solution using a novel adsorbent macrocyclic compound.

The use of macrocyclic compounds to remove organic dyes is fascinating because they have a wide surface area range and can be used for different things. new (14E, 34E)-7-Hydroxy-7, 8, 22, 23, 24, 25, 26, 27-Octahydro-6H, 16H, 33H Tetrabenzo[f,k,u,z][1,5,13,20]Tetraoxacycloheptacosine-16,33-Dione (HOTTD) was obtained by a simple high-dilution method, and characterized by FTIR, 1H-NMR, FESEM, EDX, and XRD. It worked well in removing organic dyes from aqueous solutions. Contact time, pH, dosage, initial concentration and temperature were studied. The optimum conditions were achieved by using 20 mg/L dye concentration, 50 mg dose of adsorbent and pH 9.0 at room temperature. The adsorption process was remarkably fast and reached equilibrium within 10 min for both Brilliant Green and Malachite Green while 70 min for Safranin. The batch adsorption experiments followed a pseudo 2nd order and Langmuir model with maximum adsorption capacity 19.26 mg/g, 18.28 mg/g, and 14.35 mg/g for Brilliant Green, Malachite green and Safranin respectively. The process was endothermic and spontaneous in nature. Adsorbent regeneration test provides an excellent value 5 times.

Branched poly(ethylenimine) carbon dots-MnO2 nanosheets based fluorescent sensory system for sensing of malachite green in fish samples.

Malachite green (MG) is an organic dye compound that is frequently used as a fungicide and antiseptic in aquaculture. However, human or animal exposure to MG causes carcinogenic, teratogenic and mutagenic effects. Herein, a novel fluorescent assay was designed for the detection of MG using manganese dioxide nanosheets (MnO2 NS) as an energy acceptor to quench the fluorescence of branched poly(ethylenimine) carbon dots (BPEI-CDs) via Förster resonance energy transfer. When butyrylcholinesterase is introduced to form thiocholine in the presence of S-butyrylthiocholine iodide, MnO2 NS can be recovered by thiocholine to Mn2+, resulting in restoration of the fluorescence of BPEI-CDs. Exploiting these changes in fluorescence intensity in the above system, a fluorescence probe was successfully developed for the quantitative detection of MG. Besides, this assay was applied to fish samples, verifying the high potential for practical application of the proposed sensor for the monitoring of MG in aquatic products.

Adsorption of Malachite Green (MG) as a Cationic Dye on Amberlyst 15, an Ion-Exchange Resin.

Crystals of malachite green (MG), being water-soluble, are effective agents applied to combat fungal and parasitic infections in fish. This study was conducted to evaluate the adsorption of MG as a cationic dye by polymeric resin Amberlyst 15. Changes in several parameters were observed, including the concentration of MG, pH, the adsorption rate and extent, and the temperature that might all affect the efficiency of adsorption. The adsorption method was described well by both the Redlich-Peterson and Langmuir isotherms with R 2 of 0.9933 and 0.9880, respectively. The kinetic information is consistent with the Freundlich isotherm model and pseudo-2nd-order kinetics model. Analysis of malachite green was executed by HPLC containing a Eurospher 100-5 C18 (25 cm × 4.5 mm, size of particle 5 µm) column, UV detector was set at 618 nm, and 125 mM ammonium acetate was adapted to pH 4.5 with formic acid-acetonitrile (45 : 55, v/v) as the moving phase. The limit of the discovery factor was 0.02 µgL. The negative value of ΔG° reveals the spontaneity of the absorption method. The positive value of ΔS° (333.1253 J/K mol) gives back randomness at the solid-liquid interface of sorption. The required adsorbent concentration was calculated for removing MG up to an extraction efficiency of 98.27% after 240 minutes.

Adsorption of dyes methyl violet and malachite green from aqueous solution on multi-step modified rice husk powder in single and binary systems: Characterization, adsorption behavior and physical interpretations.

A novel modified rice husk (MRH) has been prepared for removing cationic dyes in both single system and binary system. SEM-EDS, FT-IR, XRD and XPS were used to characterize the physical and chemical properties of MRH. It showed that the maximum adsorption capacity of MRH for methyl violet (MV) and malachite green (MG) in single system was 154.49 and 996.97 mg g-1, while in binary system was 530.94 and 408.58 mg g-1, respectively. The experimental results showed that the pseudo-second-order kinetic model was better to describe the kinetic behavior of MV and MG adsorption. By using double layer adsorption model, we found that the nD for MV adsorption were 2.52, 2.65 and 3.34 at 298, 308 and 318 K, respectively, and the nD for MG adsorption were 4.59, 4.85 and 4.30, respectively. These results illustrated that multiple dye molecules were adsorbed on one adsorption site in non-parallel direction, indicating that the adsorption of dyes is multi-molecular mechanism. Furthermore, synergistic and antagonistic adsorption might be existed simultaneously in binary system. In summary, MRH has been shown well adsorption properties and reusability and our finding might provide a new idea for developing low-cost, efficient and reusable adsorbent to remove dyes from wastewater.

Malachite Green, the hazardous materials that can bind to Apo-transferrin and change the iron transfer.

Different groups of synthetic dyes might lead to environmental pollution. The binding affinity among hazardous materials with biomolecules necessitates a detailed understanding of their binding properties. Malachite Green might induce a change in the iron transfer by Apo-transferrin. Spectroscopic studies showed malachite green oxalate (MGO) could form the apo-transferrin-MGO complex and change the Accessible Surface Area (ASA) of the key amino acids for iron transfer. According to the ASA results the accessible surface area of Tyrosine, Aspartate, and Histidine of apo-transferrin significantly were changed, which can be considered as a convincing reason for changing the iron transfer. Moreover, based on the fluorescence data MGO could quench the fluorescence intensity of apo-transferrin in a static quenching mechanism. The experimental and Molecular Dynamic simulation results represented that the binding process led to micro environmental changes, around tryptophan residues and altered the tertiary structure of apo-transferrin. The Circular Dichroism (CD) spectra result represented a decrease in the amount of the α-Helix, as well as, increase in the ß-sheet volumes of the apo-transferrin structure. Moreover, FTIR spectroscopy results showed a hypochromic shift in the peaks of amide I and II. Molecular docking and MD simulation confirmed all the computational findings.

Dummy Template-Based Molecularly Imprinted Membrane Coating for Rapid Analysis of Malachite Green and Its Metabolic Intermediates in Shrimp and Fish.

A novel malachite green molecularly imprinted membrane (MG-MIM) with specific selectivity for malachite green (MG) and leucomalachite green (LMG) was prepared using a hydrophobic glass fiber membrane as the polymer substrate, methyl violet as a template analog, 4-vinyl benzoic acid as the functional monomer, and ethyleneglycol dimethacrylate as the crosslinking agent. MG-MIM and non-imprinted membrane (NIM) were structurally characterized using scanning electron microscopy, surface area analyzer, Fourier-transform infrared spectrometer and synchronous thermal analyzer. The results showed that MG-MIM possessed a fluffier surface, porous and looser structure, and had good thermal stability. Adsorption properties of MG-MIM were investigated under optimal conditions, and adsorption equilibrium was reached in 20 min. The saturated adsorption capacities for MG and LMG were 24.25 ng·cm-2 and 13.40 ng·cm-2, and the maximum imprinting factors were 2.41 and 3.20, respectively. Issues such as "template leakage" and "embedding" were resolved. The specific recognition ability for the targets was good and the adsorption capacity was stable even after five cycles. The proposed method was successfully applied for the detection of MG and LMG in real samples, and it showed good linear correlation in the range of 0 to 10.0 µg·L-1 (R2 = 0.9991 and 0.9982), and high detection sensitivity (detection limits of MG and LMG of 0.005 µg/kg and 0.02 µg·kg-1 in shrimp, and 0.005 µg/kg and 0.02 µg/kg in fish sample). The recoveries and relative standard deviations were in the range of 76.31-93.26% and 0.73-3.72%, respectively. The proposed method provides a simple, efficient and promising alternative for monitoring MG and LMG in aquatic products.

Aptamer-modified magnetic SERS substrate for label-based determination of cardiac troponin I.

A sensitive label-based SERS strategy composed of magnetic bimetallic nanoparticles Fe3O4@Ag@Au, specific aptamer, and Bradford method was developed for the quantitative determination of cardiac troponin I (cTnI) in human serum. The prepared substrate with high magnetic character, signal enhancement, and uniformity exhibited significant Raman response. After the substrate was bound to the aptamer, the target protein cTnI was specifically captured, and it showed the Raman signal when the signal reporter Coomassie Brilliant Blue G-250 (CBBG) was supplied. The Raman signal intensity at 1621 cm-1 showed a wide linear relationship with the log value of the cTnI concentration in the range 0.01 to 100 ng·mL-1, and the estimated limit of detection (LOD) was 5.50 pg·mL-1. The recovery and relative standard deviation (RSD) of the spike experiment in human serum samples were 92-115% and 7.4-12.7%, respectively.

An Engineered Thermostable Laccase with Great Ability to Decolorize and Detoxify Malachite Green.

Laccases can catalyze the remediation of hazardous synthetic dyes in an eco-friendly manner, and thermostable laccases are advantageous to treat high-temperature dyeing wastewater. A novel laccase from Geothermobacter hydrogeniphilus (Ghlac) was cloned and expressed in Escherichia coli. Ghlac containing 263 residues was characterized as a functional laccase of the DUF152 family. By structural and biochemical analyses, the conserved residues H78, C119, and H136 were identified to bind with one copper atom to fulfill the laccase activity. In order to make it more suitable for industrial use, Ghlac variant Mut2 with enhanced thermostability was designed. The half-lives of Mut2 at 50 °C and 60 °C were 80.6 h and 9.8 h, respectively. Mut2 was stable at pH values ranging from 4.0 to 8.0 and showed a high tolerance for organic solvents such as ethanol, acetone, and dimethyl sulfoxide. In addition, Mut2 decolorized approximately 100% of 100 mg/L of malachite green dye in 3 h at 70 °C. Furthermore, Mut2 eliminated the toxicity of malachite green to bacteria and Zea mays. In summary, the thermostable laccase Ghlac Mut2 could effectively decolorize and detoxify malachite green at high temperatures, showing great potential to remediate the dyeing wastewater.