Tris-HCl Buffer Research Articles

Distinct functional properties of secretory l-asparaginase Rv1538c involved in phagosomal survival of Mycobacterium tuberculosis

The emergence of drug-resistant Mycobacterium tuberculosis (Mtb) stains has escalated the need for developing more efficient drugs and therapeutic strategies against tuberculosis. Here we functionally annotate a secretory mycobacterial asparaginase Rv1538c (MtA) and describe its biochemical properties. MtA primarily existed as dimer along with a minor population of multimers. Circular dichroism and fluorescence spectroscopy demonstrated a compact structure in Tris HCl buffer at pH 8.0. Under these conditions it also displayed optimum activity. It retained ∼40% activity at pH 5.5, supporting its physiological relevance in acidic phagosomal environment. MtA contravened classical Michaelis-Menten kinetics and exhibited product inhibition profile, yielding a Kcat of 869.4 s−1 and an apparent Km of 8.36 mM. We report the presence of several antigenic epitopes and a C-terminal YXXXD/E motif in MtA, hinting towards its potential to interact or influence host immune system. This was supported by our observation of morphological changes in MtA-treated human B lymphoblasts. We propose that MtA is a dual purpose enzyme used by Mtb to survive inside its host by; 1) ammonia-mediated neutralization of the phagosomal acidic pH and 2) inducing stress to primary immune cells and compromising the host immune response. Overall, this study contributes to our understanding of the biological role of mycobacterial asparaginase opening avenues for developing effective TB therapeutics.

Geomorphologic study of human tooth root surfaces following simulated avulsion by scanning electron microscopy.

Tooth avulsion is the most severe traumatic tooth injury. Immediate replantation after avulsion provides the highest success rate, whereas delayed replantation with dehydration for >1h always results in ankylosis, replacement resorption, and eventual tooth loss. The aim of this study was to investigate the geomorphologic change of ultrastructure on tooth root surfaces due to dehydration. Twenty-four sound human premolars and fully developed third molars without periodontitis were selected for the experiment. Roots were separated into 6-7 pieces with an intact root surface area (3×3mm2 ) and then divided into the following groups: fresh group, prolonged dehydration groups (air drying for 1, 2, 4, 12, or 24h), and dehydrated teeth (air drying for 2h) treated with acid (Tris-HCl buffer or citric acid buffer). More than six pieces of root from each group were subjected to scanning electron microscopy (SEM) observation. Captured images were exported to ImageJ software to quantitatively analyze the areas covered with fibers. Statistical significance was determined by comparing the means of the different groups using t-testing or one-way analysis of variance followed by post hoc testing. Fibrous "vegetation" covering the cementum was observed on the fresh root surface by SEM. This was destroyed by dehydration (>1h), resulting in a root surface resembling the "Gobi Desert." The difference was statistically significant (p<.001). Root surface deteriorated by dehydration could be recovered by the re-use of the fibers embedded in the cementum as acid demineralization of the outer layer of cementum exposed the embedded fibers to simulate the geomorphology of fresh root surface. Dehydrated teeth had deteriorated geomorphology of the root surface, which could be reversed by the re-use of the fibers embedded in the cementum using citric acid. Direct evidence from SEM gives new insights into the replantation of dehydrated avulsed teeth.

Fluorescent Visualization of Nucleolar G-Quadruplex RNA and Dynamics of Cytoplasm and Intranuclear Viscosity

The nucleolus, the locus of ribosome biogenesis, was found to be the predominant intracellular target of a new fluorescent probe, V-P1. In solution, the probe demonstrated both a selectivity to RNA...

OPTIMISATION OF AN ELECTROCHEMICAL SENSOR BASED ON BARE GOLD ELECTRODE FOR DETECTION OF ALUMINIUM ION

In this work, an electrochemical method for detection of trace amount of aluminium (Al3+), a heavy metal ion, based on a bare gold electrode (AuE) was developed. Current responses of the AuE under various type of electrolytes, redox indicators, pH, scan rate and accumulation time were investigated using cyclic voltammetry (CV) method to obtain the optimum conditions for Al3+ detection. The sensing properties of the AuE towards the target ion with different concentrations were investigated using differential pulse voltammetry (DPV) method. From the CV results, the optimalconditions for the detection of Al3+ were Tris-HCl buffer (0.1 M, pH 2) supported by 5 mM Prussian blue with scan rate and accumulation time respectively of 100 mVs−1 and 15 s. Under the optimum conditions, the DPV method was detected with different concentrations of aluminium ion ranging from 0.2 to 1.0 ppm resulted in a good linear regression r² = 0.9806. This result suggests that the optimisation of the basic parameters in electrochemical detection using AuE is crucial before further modification of the Au-electrode to improve the sensitivity and selectivity especially for the low concentration of ion detection. The developed method has a great potential for rapid detection of heavy metal ion (Al3+) in drinking water samples.

Pressurized water extraction – the fast and efficient method for isolation of bioactive proteins from Viscum album leaves

New strategies for the fast, efficient, and environmentally friendly extraction of proteins are required to isolate desired bioactive compounds from a technological point of view. In this study, utilization of the pressurized water extraction (PWE) at low temperature (40 °C) for isolation of mistletoe proteins was investigated. PWE effectiveness, based on protein fingerprints, were compared with those obtained by conventional extractions using 10 mmol L–1 Tris-HCl buffer pH 8.3, 50 mmol L–1 phosphate buffer pH 7, or deionized water. The extracts were precipitated using acetone, trichloroacetic acid (TCA), and 20% (w/v) TCA/acetone and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PWE was more or equally efficient for isolation of mistletoe proteins than evaluated conventional extraction methods. The proteomic analysis combining mass spectrometry and database searching confirmed the presence of 35 proteins in PWE extracts precipitated by acetone, which was the most compounds identified from all studied extracts. The PWE high extraction power was revealed for multiple viscotoxin isoforms and specific enzymes indispensable for the synthesis of terpenes.

A novel phenanthroline-based colorimetric turn-off optical sensor for Zn2+

In this study, a new Zn2+-selective sensor (PCA) was developed by linking 4-(dimethylamino) cinnamaldehyde with 2,9-dimethyl-1,10-phenanthroline in a one-step reaction. The colorimetric and optical sensing properties of PCA were investigated by utilizing UV–Vis, fluorescence spectroscopy and the naked eye. The sensor PCA displayed high selectivity and sensitivity to Zn2+ over a wide range of the other metal ions in acetonitrile (ACN)/Tris-HCl buffer (10 mM, pH 7.39, v/v 2:1) solution. The sensor showed a clear visual change from yellow to orangish-red with Zn2+. In addition, PCA could detect Zn2+ with fluorescence quenching. The complex (PCA/Zn2+) stoichiometry was found to be 1:1 and the binding constant of the complex was calculated as 1.30 × 106 M−1. PCA displayed a low detection limit (0.835 µM) which is much lower than the value recommended by WHO. PCA was also used to detect Zn2+ in tap water samples using filter paper strips and UV–Vis spectroscopy.

An in-gel digestion method of chymotrypsin to improve sequence coverage of membrane protein by mass spectrometry

In recent years, mass spectrometry has been widely used to study membrane protein structure and function. However, the application of mass spectrometry to study integral membrane protein is limited because there are many hydrophobic amino acids in the trans-membrane domain of integral membrane protein to cause low sequence coverage detected by LC-MS/MS. Therefore, we used vitamin K epoxide reductase (VKORC1), a human integral membrane protein, as a model to optimize the digestion conditions of chymotrypsin, and developed an in-gel digestion method of chymotrypsin to improve sequence coverage of membrane protein by mass spectrometry. By exploring the effects of calcium concentration, pH value and buffer system on the percentage of sequence coverage, number of total detected and types of unique peptide, and the size of unique peptide, sequence coverage and peptide diversity could be considered under condition of Tris-HCl buffer with 5-10 mmol/L calcium ion concentration and pH value 8.0-8.5. This method could make the sequence coverage of membrane protein to reach more than 80%. It could be widely used in the study of membrane protein structure and function, identification of interaction site between membrane proteins, and identification of binding site between membrane protein and small molecular drug.

A cost effective SiO2-CaO-Na2O bio-glass derived from bio-waste resources for biomedical applications.

The present paper describes the in vitro bioactivity, cytocompatibility and degradation performance of SiO2-CaO-Na2O bio-glass synthesized using bio-waste. Egg shells and rice husk ash (RHA) bio-wastes were used as sources of calcium oxide (CaO) and silica (SiO2), respectively. Glass samples were obtained by melt-quenching technique. Bioactivity was studied using in vitro experiments in simulated body fluid (SBF), degradation behaviour was evaluated in Tris-HCl buffer solutions recommended by ISO 10993-14 standards and cytocompatibility was estimated using MTT assay. The formation of hydroxyapatite was characterized by XRD, FTIR and SEM-EDS after soaking the glass samples in SBF solution. XRD confirmed the phase of hydroxyapatite with its standard JCPDS data. FTIR analyses revealed the occurrence of distinctive functional groups related to hydroxyapatite. Surface micrographs showed the agglomerated globular shape morphology of hydroxyapatite, while EDS analysis confirmed the existence of biological elements of apatite such as Ca, P and O. Degradation study results showed that the glass thus prepared has considerable controlled degradation rate. MTT assay revealed the cytocompatibility nature for different dosages (1000-50μg/mL) of the prepared glass with MG-63 cells. These results perfectly established that egg shells and RHA are potentially beneficial resources for the production of bio-glasses.

Optimization and Analytical Behavior of Electrochemical Sensors Based on the Modification of Indium Tin Oxide (ITO) Using PANI/MWCNTs/AuNPs for Mercury Detection.

In the present study, indium tin oxide (ITO) was used as a transparent working electrode for the development of an electrochemical sensor for the detection of mercury (II) ions (Hg2+). The electrode was modified by direct electrodeposition of polyaniline (PANI), multiwalled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) followed by optimization of the analyte and operating conditions, aiming to improve the selectivity, sensitivity and reliability of the electrode for mercury detection. Successful immobilization of the PANI and nanomaterials (MWCNTs and AuNPs) on the ITO electrode was confirmed by Scanning Electron Microscope (SEM), Energy Dispersive X-ray (EDX) and Fourier Transform Infrared Spectroscopy (FTIR) analyses. The optimum conditions for mercury detection using the modified ITO electrode were pH 7.0 of Tris-HCl buffer (50 mM) in the presence of 1 mM methylene blue (MB) as a redox indicator, a scan rate of 0.10 V·s−1 and a 70 s interaction time. The electrochemical behavior of the modified electrode under the optimized conditions indicated a high reproducibility and high sensitivity of mercury detection. It is therefore suggested that the PANI/MWCNT/AuNP-modified ITO electrode could be a promising material for the development of on-site mercury detection tools for applications in fields such as diagnostics, the environment, safety and security controls or other industries.

Behavior of Araujiain, a new cysteine phytoprotease, in organic media with low water content

In this paper we studied the effect of different organic solvents (1-octanol, trichloroethylene, ethanol, ethyl acetate, tetrahydrofuran, cyclohexane, propanone, acetonitrile, dichloromethane, chlorobenzene, N,N-dimethylformamide, acetophenone, diethyl ether, methanol, ethylene glycol and toluene) with low and constant water content on substrate preferences, thermostability and stability (caseinolytic activity retention after 4 h) of proteases of Araujia hortorum Fourn. ( Asclepiadaceae ). The stability of araujiain was high in N,N-dimethylformamide and ethanol at 40oC, but decreased at higher temperature. Araujiain substrates preferences in buffer Tris-HCl (pH 8), ethylene glycol and N,N-dimethylformamide exhibited different patterns, but the enzyme showed a high preference by glutamine derivative in all cases. According to FTIR spectroscopy studies, araujiain changed its secondary structure and as a consequence, it also changed its substrate preferences. This enzyme showed lower α-helical character and greater β-sheet folding in buffer than in organic media. A larger amount of antiparallel β-sheet residues indicates the formation of tighter intermolecular hydrogen bonds and enzymatic aggregates. These facts could explain the higher esterolytic activities, the greater stability and good hydrolytic potential of araujiain in some organic media such as N,N-dimethylformamide.

Biophysical characterization of the structural stability of Helix lucorum hemocyanin

The structural stability of the hеmocyanin purified from the hеmolymph of gаrden snаils Helix lucorum (HlH) was investigated by means of far-UV circular dichroism (CD), differential scanning calorimetry (DSC) and transmission electron microscopy (TEM). For the first time, TEM analyses showed the presence of tubular polymers in hemocyanins after three-day dialysis against a stabilizing buffer containing high concentrations of Ca2+ and Mg2+ ions (100 mmol L −1). The conformational stability study of native HlH by means of CD in a wide pH range (2.5-11.5) defined the pH stability region of HlH at pH 6.5 − 8.0. DSC analyses demonstrated the thermal stability of this hemocyanin. One transition, with an apparent transition tеmperature (T m) at 82.3 °C, was detected in the heаt capacity curve of HlH in 50 mmol L −1 Tris-HCl buffer, pH 7.2, at a heating rate of 1.0 °C min−1. The calorimetrically observed thermal transition correlates well with the unfolding transition monitored by CD measurements. The two-state kinetic model was used to analyse the process of irreversible thermal denaturation of HlH; Е a of 451 ± 4 kJ mol−1 was calculated. The obtained results on the conformational stability of HlH will facilitate the further investigation of the properties and potential biomedical applications of this respiratory protein.

Enhancing immobilization of Aspergillus oryzae ST11 lipase on polyacrylonitrile nanofibrous membrane by bovine serum albumin and its application for biodiesel production

The partially purified lipase from Aspergillus oryzae ST11 was immobilized on the surface-modified electrospun polyacrylonitrile (PAN) nanofibrous membrane. The effects of time and concentrations of glutaraldehyde and bovine serum albumin (BSA) were studied. The immobilized lipase reached the maximum activity at 180 min with the recovered activity of 44.2% (0.22 U/mg-support) in the absence of BSA. After adding 6 mg/mL BSA and 40 mM glutaraldehyde gave the recovered activity at 86.9% (0.43 U/mg-support). It showed the highest stability at pH 7.5 but the soluble lipase was at pH 7.0. In addition, the immobilized lipase was more stable at pH 4.0 and 9.0. The immobilized lipase preserved 56% activity at 70 °C but the soluble lipase had 34% activity. It conserved 86% activity while the soluble lipase had 18% activity after 13 days of storage at 4 °C. The operating parameters such as biocatalyst concentration, water content, and the molar ratio of methanol and palm oil were optimized for biodiesel production. The highest biodiesel conversion (95%) was obtained with the immobilized lipase 15% (w/w), 40% (w/w) 50 mM Tris-HCl buffer pH 7.0 and methanol/palm oil (3.5:1.0). This immobilized lipase could be reused with 82.9% conversion after 10 cycles of batch production.

Activity profile of innate immune-related enzymes and bactericidal of freshwater fish epidermal mucus extract at different pH

The epidermal mucus of fish performs diverse functions from prevention of mechanical abrasion to limit pathogen invasions. The current experiment was designed to extract skin mucus proteins of three freshwater fish, i.e. common carp (Cyprinus carpio), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita) with organic solvent (methanol) and dissolve in different pH of Tris-HCl buffers to examine the significance of pH in the solubilisation of skin mucus proteins. The protein profiles of different pH solubilised methanol fish skin mucus extracts were determined by SDS-PAGE. The non-specific immune enzymes, alkaline phosphatase, lysozyme and protease of fish skin mucus were compared and this present study demonstrated that these enzymes differed in their activity depending on pH buffers. The higher lysozyme and protease activity were observed at the pH of 8.0 and higher alkaline phosphatase activity in the pH 9.0 of C. mrigala fish skin mucus methanol extract. In addition, the bactericidal activity was evaluated against the pathogens Proteus vulgaris and Pseudomonas aeruginosa. The pH 8.0 of C. mrigala skin mucus extract revealed better bactericidal activity than other fish species mucus pH buffers against both P. vulgaris and P. aeruginosa. In the case of protein profile from SDS-PAGE, based on pH buffers and the solubilisation of proteins, differences in the resolution of bands were observed. The higher alkaline pH of 9.0 showed smeared gel bands in all the three fish skin mucus methanol extract. The present study suggests that methanol extracted C. mrigala fish skin mucus at pH 8.0 showed better innate immune enzymes and bactericidal activity. The additional examinations of C. mrigala skin mucus methanol extract in this pH aids in identifying novel bioactive molecules. This is the study of proteome of three fish species skin mucus in the effect of pH. Further analyses are required to evaluate proteins present in fish skin mucus extracted with methanol and the influence of pH on protein solubility. These findings could be helpful in exploring natural alternatives to antibiotics in aquaculture industry against infectious pathogens.

Effect of silicon dioxide nanoparticles on the assessment of quercetin flavonoid using Rhodamine B Isothiocyanate dye

Quercetin is a plant flavonol from the flavonoid group of polyphenols; it has medical importance in treating many diseases. Quercetin flavonoid was assessed using Rhodamine B Isothiocyanate (RBITC) dye in absence and presence of SiO2 nanoparticles at pH = 7 using Tris-HCl buffer solution. This method considered a simple, sensitive, and selective method where the dye has fluorescence emission band at 583 nm after excitation at 552 nm which strongly quenched by quercetin. The results showed that presence of SiO2 nanoparticles which were characterized using different techniques such as XRD, SEM, EDX, IR, TGA and TEM analysis has a significant effect on lowering the detection limit for quercetin using RBITC dye from 19.99 to 9.86 nmol.L−1, increasing the binding constant from 7.17 × 103 to 25.92 × 103 mol−1.L in absence and presence of SiO2 nanoparticles respectively. The interaction between quercetin and RBITC dye was of hydrophobic type and the fluorescence quenching mechanism was static in absence and presence of SiO2 nanoparticles respectively. The method was applied for the determination of quercetin in spiked human plasma where recovery measurements proved the accuracy of this method.

Large-scale direct pyrolysis synthesis of excitation-independent carbon dots and analysis of ferric (III) ion sensing mechanism

A one-step direct pyrolysis method without any post-treatment steps and a mass yield of up to 69% was realized for preparing N-doped carbon dots (NCDs). The NCDs exhibit excellent excitation-independent blue fluorescence with quantum yield (QY) of 65.5%, and their luminescence mechanism was explored based on optical properties and structural composition, which mainly originates from surface state photoluminescence (PL). The NCDs display highly selective and sensitive detection of Fe3+ in tris-HCl buffer system, a good linear relationship between PL intensity ratios and Fe3+concentrations was obtained in concentration range of 0 to 400 μM with a low LOD of 0.703 μM. The formation of ferric hydroxide colloid is the main reason for the specific recognition of Fe3+ by NCDs, and buffer solution plays a key role in the formation of ferric hydroxide colloid and selective detection of Fe3+. The sensing mechanism is a dynamic quenching electron transfer process in the action of hydrogen bonding and colloidal adsorption. Three real water samples were applied to evaluate actual application of NCDs probe, satisfactory recoveries (93.5%–105.6%) indicate that NCDs has great practical potential for detection of metal ions in environmental water assays.

Fabrication of fluorescent hybrid nanomaterials based on carbon dots and its applications for improving the selective detection of Fe (III) in different matrices and cellular imaging.

Considering that detection on cations or ions still meets some challenges in achieving the effectivity and selectivity just by employing one platform, the ingenious fabrication of nanomaterials exhibits an increasing research interests for the preponderance in improving or integrating the performance of single platform. Herein, a fluorescent hybrid nanomaterials based on an organic dye 4-methylumbelliferone (4-MU) as modifier and D-arginine as carbon cores has been developed via a facile one-step hydrothermal synthesis, forming carbon dots (CDs)/4-MU hybrid nanomaterials (CDs-4-MU). This kind of nanomaterials can improve the sensitive and selective detection of single CDs towards Fe3+ ions in different matrices. The detection mechanism of CDs-4-MU towards Fe3+ can be attributed to an electron transfer process between CDs-4-MU and Fe3+, leading to the fluorescence quenching. The limit of detection (LOD) and corresponding linear range in tris-HCl buffer solution are 0.68μM and 2.29-200μM, respectively. Furthermore, this nanomaterial can also achieve a detection of Fe3+ ions in real samples such as tap water, culture medium and fetal bovine serum. In particular, CDs-4-MU exhibits a good biocompatibility and can be uptaken by MC3T3 cells, thus can be applied for Fe3+ ions detection in cellular level and cellular imaging. Therefore, this work provides a versatile strategy for the synthesis of CDs-based hybrid nanomaterials and opens a new pathway for improving the ion detection in real samples, which is of significance in practical applications.

Extraction and Purification of Lipases Enzyme from Germinating Seeds of Four Crops

Lipase enzyme has attracted a lot of attention in recent years because of its diverse biotechnological applications. The present study was conducted to screen germinated seeds of four crops, namely sunflower (Helianthus annuus), flaxor linseed (Linum usitatissimum ), peanut (Arachis hypogaea ) and castor bean (Ricinus communis), for the activity of their lipases. to the study also included the extraction and purification of lipase from the seeds of the most promising crop using different solvents.&#x0D; The results indicated that the maximum enzymatic activity (0.669 U/ml) was obtained when 0.1 M Tris-HCl buffer extract was used after 3 days of seed germination of all the tested species, as compared to the other test solvents (acetone and water). Sunflower germinated seeds showed the highest lipase activity, which was higher by 159.67, 185.32, and 285.90 % over the activities of castor bean, flax, and peanut seeds, respectively. Among the used ranges of saturation of ammonium sulfate, the ratio of 70% was the best in precipitating the crude enzyme, showing a highest specific activity of 2.576 U/ mg protein.&#x0D; The first stage of gel filtration chromatography column by Sephadex G-200 indicated the presence of two non-identical peaks, one for protein and another for lipase activity, between the fractions of 18 to 23. The active fractions were pooled and loaded again in the Sephadex G-200 column and the eluted fractions showed two identical peaks, one for protein and another for lipase activity, between the fractions of 19 to 23. The final purification step by gel filtration showed a specific activity of 6.482 U/mg proteins with a yield of 38.75 % and 11.33 folds of time of purification. The study revealed that sunflower seeds are a better source of lipase as compared to the other used plant seeds, which can be used in different industries.

Characterization of a novel hyper-thermostable and chlorpyrifos-hydrolyzing carboxylesterase EstC: A representative of the new esterase family XIX

Carboxylesterases have widely been used in a series of industrial applications, especially, the detoxification of pesticide residues. In the present study, EstC, a novel carboxylesterase from Streptomyces lividans TK24, was successfully heterogeneously expressed, purified and characterized. Phylogenetic analysis showed that EstC can be assigned as the first member of a novel family XIX. Multiple sequence alignment indicated that EstC has highly conserved structural features, including a catalytic triad formed by Ser155, Asp248 and His278, as well as a canonical Gly-His-Ser-Ala-Gly pentapeptide. Biochemical characterization indicated that EstC exhibited maximal activity at pH 9.0 (Tris-HCl buffer) and 55 °C. It also showed higher activity towards short-chain substrates, with the highest activity for p-nitrophenyl acetate (pNPA2) (Km = 0.31 ± 0.02 mM, kcat/Km = 1923.35 ± 9.62 s−1 mM−1) compared to other pNP esters used in this experiment. Notably, EstC showed hyper-thermostability and good alkali stability. The activity of EstC had no significant changes when it was incubated under 55 °C for 100 h and reached half-life after incubation at 100 °C for 8 h. Beyond that, EstC also showed stability at pH ranging from 6.0 to 11.0 and about 90% residual activity still reserved after treatment at pH 8.0 or 9.0 for 26 h, especially. Furthermore, EstC had outstanding potential for bioremediation of chlorpyrifos-contaminated environment. The recombinant enzyme (0.5 U mL−1) could hydrolyze 79.89% chlorpyrifos (5 mg L−1) at 37 °C within 80 min. These properties will make EstC have a potential application value in various industrial productions and detoxification of chlorpyrifos residues.

Adaptation of two-dimensional electrophoresis for muscle tissue analysis

It is important to understand the molecular mechanisms that take place in muscle tissues and to predict meat quality characteristics. One of the most popular methods is two-dimensional electrophoresis, which allows us to visualize, share and identify different molecules, including meat proteins. However, the standard conditions of this method are not universal for all types of raw material, so the authors suggest a new variation of two-dimensional electrophoresis for muscle tissue analysis. Samples were tested by the classical version of isoelectric focusing (cathode buffer in the top and anode buffer in the bottom chamber of the electrophoresis cell) and its variation (anode buffer in the top and cathode buffer in the bottom chamber of the electrophoresis cell). Next, extruded gels were incubated in two different buffer systems: the first was equilibration buffer I (6 M urea, 20% w/v glycerol, 2% w/v SDS and 1% w/v Ditiothreitol in 375 mM Tris-HCl buffer, pH 8.8) followed by equilibration buffer II (6 M urea, 20% w/v glycerol, 2% w/v SDS and 4% w/v iodoacetamide in 375 mM Tris-HCl buffer pH 8.8 and the second, buffer А, consisting of 5 M urea, 2% w/v SDS, 5% v/v mercaptoethanol, 62.5 mM Tris-HCl buffer, pH 6.8 and 0.01% w/v bromophenol blue. Electrophoretic studies of muscle tissue revealed the best protein separation after changing the direction of the current (authors' variation), while no differences were detected after changing incubation buffers.

Environment-friendly treatment to reduce photoyellowing and improve UV-blocking of wool

Wool products are widely used in people's daily life due to their various excellent properties. However, long-term exposure to solar radiation containing ultraviolet light (UV) can cause severe photoyellowing issues. Also, when used to fabricate apparel, fabrics are expected to own an outstanding UV-blocking feature to protect the skin underneath. In this study, we have developed an eco-friendly treatment to reduce the photoyellowing as well as improve UV-blocking capacity of wool fabrics by introducing para-aminobenzoic acid (PABA) through a microbial transglutaminase (mTGase) mediated acyl transfer reaction in a tris(hydroxymethyl)aminomethane-hydrochloric acid (Tris-HCl) buffer at mild condition. Fluorescent and UV–Vis reflectance spectra proved the successful attachment of PABA on wool fabrics. In addition, relative properties measured after laundering demonstrated the existence of strong covalent bonds, which ensure the stable performance of corresponding fabrics. Moreover, we found that wool fabrics processed by an appropriate concentration of PABA in the enzyme-tris reaction system could effectively resist photoyellowing and exhibit dramatic improvement of UV protection factor (UPF) values. Tensile test results indicated some improvement in the mechanical performance of the treated fabrics after long-term accelerated irradiation. The feasible and effective method developed in current research has great potential to improve the durability and performance of wool products.