Addition Of Detergent Research Articles

Heterologous expression and characterization of functional mushroom tyrosinase (AbPPO4)

Tyrosinases are an ubiquitous group of copper containing metalloenzymes that hydroxylate and oxidize phenolic molecules. In an application context the term ‘tyrosinase’ usually refers to ‘mushroom tyrosinase’ consisting of a mixture of isoenzymes and containing a number of enzymatic side-activities. We describe a protocol for the efficient heterologous production of tyrosinase 4 from Agaricus bisporus in Escherichia coli. Applying this procedure a pure preparation of a single isoform of latent tyrosinase can be achieved at a yield of 140 mg per liter of autoinducing culture medium. This recombinant protein possesses the same fold as the enzyme purified from the natural source as evidenced by single crystal X-ray diffraction. The latent enzyme can be activated by limited proteolysis with proteinase K which cleaves the polypeptide chain after K382, only one The latent enzyme can amino acid before the main in-vivo activation site. Latent tyrosinase can be used as obtained and enzymatic activity may be induced in the reaction mixture by the addition of an ionic detergent (e.g. 2 mM SDS). The proteolytically activated mushroom tyrosinase shows >50% of its maximal activity in the range of pH 5 to 10 and accepts a wide range of substrates including mono- and diphenols, flavonols and chalcones.

Lactobacillus brevis Lipase: Purification, Immobilization onto Magnetic Florosil NPs, Characterization and Application as a Detergent Additive

Abstract In this study, a thermo-tolerant and alkaline lipase enzyme was purified from Lactobacillus brevis and immobilized onto modified γ-Fe3O4 florisil nanoparticles (γ-Fe3O4 MF NFs) and the usability of free lipase (FL) and immobilized lipases (IML) as detergent additives was investigated. Lipase enzyme was purified by fractional precipitation using 20% ammonium sulfate, DEAE-Sephadex ion-exchange chromatographic column, and Sephacryl S200 gel filtration chromatographic techniques. Then, the enzyme was purified, which resulted in 135.2-fold purification. Its molecular mass was determined to be 57 kDa by SDS-PAGE. The covalent immobilization of purified lipase was done using γ-Fe3O4 MF NPs. γ-Fe3O4 MF NPs and IML were characterized by using SEM, TEM, FT-IR, and XRD. IML showed a good thermo-stability and its activities were calculated as 80% at 60°C. The free and IML enzymes were most stable at alkaline pHs in the range of 7.0–10.0. Also, IML is more stable towards metal ions compared to free lipase enzyme. Washing performances of some detergent formulations were investigated in the presence and absence of Lipase. Olive oil was removed by the detergent alone and by the detergent and IML at ratios of 45% and 72%, respectively. The study on removal of oil stain from cotton cloths indicated that the removal of oil was superior in the presence of IML and IML with detergent, when compared to the detergent alone.

Optimization of the Helmintex method for schistosomiasis diagnosis

A diagnostic test that is reliable, sensitive, and applicable in the field is extremely important in epidemiological surveys, during medical treatment for schistosomiasis, and for the control and elimination of schistosomiasis. The Helmintex (HTX) method is based on the use of magnetic beads to trap eggs in a magnetic field. This technique is highly sensitive, but the screening of fecal samples consumes lots of time, thus delaying the results, especially in field studies. The objective of this work was to determine the effects of incorporation of the detergent Tween-20 into the method in an attempt to decrease the final pellet volume produced by the HTX method as well as the use of ninhydrin to stain the Schistosoma mansoni eggs. We showed that these modifications reduced the final volume of the fecal sediment produced in the last step of the HTX method by up to 69% and decreased the screening time to an average of 10.1 min per sample. The use of Tween 20 and ninhydrin led to a high percentage of egg recovery (27.2%). The data obtained herein demonstrate that the addition of detergent and the use of ninhydrin to the HTX process can optimize the screening step and also improve egg recovery, thus justifying the insertion of these steps into the HTX method.

Chemically active oil filter to develop detergent free bio-based lubrication for diesel engine

In diesel engine lubricants, over-based detergent additives are typically used to neutralize acids. These additives are required for long service life, but their use results in hazardous emissions, which mainly include sulfur and ash content. To address this problem, this study investigates the detergent-free bio-based lubricant conditioning approach by using a chemically active oil filter. A standard filter element impregnated with sodium oxide particles was used to improve lubricant quality. In this research, palm trimethylolpropane ester was synthesized for use as the base oil for formulation of a detergent-free bio-based lubricant. A condition-based approach, in which two engine endurance tests were carried out, was adopted. Lubricant samples from new and engine-aged conditions were tested for viscosity, total acid number, total base number, and corrosion. Piston ring-cylinder sliding tests were conducted to determine the wear protection and friction behavior of lubricant samples. A comparative analysis showed that lubricant conditioning by a modified filter improved the alkaline reserve and controlled the viscosity increase, thus extending the bio-based lubricant service life from 80 h to 200 h. The bio-based lubricant conditioned by a chemically active filter exhibited reduced cylinder liner wear and friction coefficient by 9.2% and 12.9%, respectively.

Bioconversion of 16-dehydropregnenolone Acetate to Exclusively 4-androstene-3,17-dione by Delftia acidovorans MTCC 3363.

Delftia acidovorans MTCC 3363 was found to convert 16-dehydropregnenolone acetate (16-DPA) exclusively to 4-androstene-3, 17-dione (AD). Addition of 9α-hydroxylase inhibitors was not required for preventing the accumulation of byproducts. The effect of pH, temperature, substrate concentration, surfactants and carrier solvents on this bioconversion has been studied. 16-DPA was maximally converted in buffered medium at pH 7.0, at temperature 30°C and 0.5 mg ml-1 substrate concentration. Detergent addition and temperature above 35°C had deleterious effect on bioconversion. Dioxan was found to be the best carrier solvent for biotransformation of 16-DPA to AD.

Changing Petroleum Based Fuels for Fuel Cell Vehicles: Composition-Execution Relationships

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Alkaline serine proteases from Helicoverpa armigera: potential candidates for industrial applications.

We characterized trypsin- and chymotrypsin-like serine alkaline proteases from cotton bollworm, Helicoverpa armigera, for their probable potential application as additives in various bio-formulations. Purification was achieved by using hydroxylapatite, DEAE sephadex and CM sephadex columns, which resulted in increased enzyme activity by 13.76- and 14.05-fold for trypsin and chymotrypsin, respectively. Michaelis-Menten constants (Km ) for substrates of trypsin and chymotrypsin, BApNA and SAAPFpNA, were found to be 1.25 and 0.085 mM, correspondingly. Fluorescent zymogram analysis indicated the presence of five trypsin bands with molecular masses of ∼21, 25, 38, 40, and 66 kDa and two chymotrypsin bands with molecular masses of ∼29 and 34 kDa in SDS-PAGE. The optimum pH was 10.0 and optimum temperature was 50°C for proteolytic activity for the purified proteases. The proteases were inhibited by synthetic inhibitors such as PMSF, aprotonin, leupeptin, pefabloc, and antipain. TLCK and TPCK inhibited about 94 and 90% of trypsin and chymotrypsin activity, respectively, while EDTA, EGTA, E64, pepstatin, idoacetamide, and bestatin did not affect the enzymes. The purified enzymes exhibited high stability and compatibility with metal ions; oxidizing, reducing, and bleaching agents; organic solvents; and commercial detergents. Short life cycles, voracious feeding behavior, and production of multiple forms of proteases in the midgut with rapid catalytic activity and chemostability can serve H. armigera as an excellent alternative source of industrially important proteases for use as additives in stain removers, detergents, and other bio-formulations. Identification of enzymes with essential industrial properties from insect species could be a bioresource.

An Original Halo-Alkaline Protease from Bacillus halodurans Strain US193: Biochemical Characterization and Potential Use as Bio-Additive in Detergents

Over a hundred of halophilic/halotolerant microorganisms were screened for alkaline protease production. The bacterium showing the highest enzyme production was characterized and identified as Bacillus halodurans US193 on the basis of 16S rRNA gene analysis. It was alkalophilic, thermophilic and halotolerant since it grew optimally at pH 9.7 and 50 °C with tolerance of up to 125 g NaCl l−1. The alkaline protease was purified 4.9 times with about 40186.1 U/mg as specific activity. It exhibited optimal activity at pH 10, 70 °C and 0.25 M NaCl with perfect stability at wide ranges of pH (6–12), temperatures (30–60 °C) and NaCl concentrations (0–2 M). The serine alkaline protease maintained high stability in the presence of Cu2+, Mg2+, Ba2+ and Ca2+ ions, various organic solvents [50% (v/v)] and ionic and non ionic detergent additives. In addition, it was more compatible with various commercialized detergents than other reported detergent proteases, and was very efficient in blood stain removal. These findings let B. halodurans US193 alkaline protease be an ideal candidate for many industrial processes at harsh conditions, especially as a bio-additive in detergent industry.

Detection and quantification of anionic detergent (lissapol) in milk using attenuated total reflectance-Fourier Transform Infrared spectroscopy.

Adulteration of milk to gain economic benefit is rampant. Addition of detergent in milk can cause food poisoning and other complications. Fourier Transform Infrared spectroscopy was evaluated as rapid method for detection and quantification of anionic detergent (lissapol) in milk. Spectra of pure and artificially adulterated milk (0.2-2.0% detergent) samples revealed clear differences in wavenumber range of 4000-500cm-1. The apparent variations observed in region of 1600-995 and 3040-2851cm-1 corresponds to absorption frequencies of common constituents of detergent (linear alkyl benzene sulphonate). Principal component analysis showed discrete clustering of samples based on level of detergent (p⩽0.05) in milk. The classification efficiency for test samples were recorded to be >93% using Soft Independent Modelling of Class Analogy approach. Maximum coefficient of determination for prediction of detergent was 0.94 for calibration and 0.93 for validation, using partial least square regression in wavenumber combination of 1086-1056, 1343-1333, 1507-1456, 3040-2851cm-1.

Characterization of thermo-solvent stable protease from Halobacillus sp. CJ4 isolated from Chott Eldjerid hypersaline lake in Tunisia.

About 110 newly isolated halophilic and halotolerant bacteria were screened for protease production. A moderately halophilic strain (CJ4), isolated from Chott Eldjerid Hypersaline lake in Tunisia, showed the highest activity on agar plate and was then selected. The biochemical and physiological characterization of the isolate along with the 16S rRNA sequence analysis placed it in the genus Halobacillus. Protease production was maximal at 120 g/L NaCl (2 M) and it started from the post-exponential phase reaching a maximum level at the early decline phase of bacterial growth. Protease activity was optimal at 0.4 M NaCl, pH 9 and 45 °C. It showed an excellent stability over wide ranges of temperatures (30-60 °C), NaCl concentrations (0-5 M), and pH values (5-10), which make it a good candidate for industrial applications at harsh conditions. Crude protease was strongly inhibited by PMSF revealing the dominance of serine proteases. Protease activity exhibited high stability in the presence of several organic solvents and detergent additives. These findings make Halobacillus sp. CJ4 protease with a great interest for many biotechnological applications at high salt or low water content such as peptide synthesis and detergent formulation.

Interaction of nonionic detergents with the specific sites of lysozyme amyloidogenic region − inhibition of amyloid fibrillization

Two nonionic detergents, Triton X-100 (TX-100) and n-dodecyl-β-d-maltoside (DDM) were tested for their ability to affect lysozyme amyloid aggregation. We have demonstrated that fibrillization of lysozyme is completely inhibited by low sub-micellar concentrations of both of these detergents. The apparent IC50 values were calculated to be 22μM and 26μM for TX-100 and DDM, respectively. The detergent/protein ratio is not the only parameter controlling inhibition. The precise timing of the detergent addition was found to be also crucial. It appears that the primary inhibitory activity of detergents resulted from inhibition of nuclei formation, in addition to inhibition of fibril polymerization at the early stage of protofibrils growth. The docking study revealed that Asn-59, Trp-63 and Ala-107, all present within the lysozyme amyloidogenic region, were involved in the interaction with both detergents. In addition, TX-100 also interacted with Gln-57 and Asp-103 within lysozyme. Moreover, based on our computational results, TX-100 bridges the Gln-57 and Ala-107 amino acids of the amyloidogenic segment of lysozyme and therefore inhibits more effectively the amyloid fibril formation. Along these lines, the knowledge gained from our study indicates that the detergents or their derivatives may be applicable as a promising strategy for the modulation of lysozyme protein aggregation.

Isolation and mass spectrometry analysis of urinary extraexosomal proteins

The aim of the present study was to develop a LC-MS/MS-based proteomic analysis method of urinary exosomal proteins that has the potential to discover disease biomarkers. In short, urinary exosomes from healthy subjects were isolated by immunocapture on magnetic beads, detected by immunofluorescence and TEM, trypsin digested directly on the beads for an accelerated time with no addition of detergents before performing an LC-MS analysis of the trypsinate. To our knowledge, this is the first proteomic analysis of proteins displayed on the outer surface of exosomes. The outer exosome proteome may contain proteins that are of higher biomarker value compared to soluble cargo protein as the proteins projecting into the extracellular milieu might be more directly involved in physiological functions of exosomes. The proteomic analysis identified 49 proteins that were considered significant; the majority is involved in carbohydrate and lipid metabolism or in immune responses. Thirty of the proteins are linked to diseases. The developed proteomic method exploiting urinary exosomes might be of great value in search for diagnostic or prognostic biomarkers of especially metabolic and immune-related diseases.

Characterization of the depolymerizing activity of commercial lipases and detection of lipase-like activities in animal organ extracts using poly(3-hydroxybutyrate-co-4-hydroxybutyrate) thin film.

Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] is one of the polyhydroxyalkanoate (PHA) copolymers which can be degraded by lipases. In this study, the depolymerizing activity of different known commercial lipases was investigated via microassay using P(3HB-co-92 mol % 4HB) thin film as substrate. Non-enzymatic hydrolysis occurred under conditions in which buffers with pH 12 and 13 were added or temperature of 50 °C and above. Different concentrations of metal ions or detergents alone did not cause the film hydrolysis. The depolymerizing activity of lipases on P(3HB-co-4HB) was optimum in the pH range of 6–8 and at temperatures between 30 and 50 °C. Addition of metal ions and detergents in different concentrations was also shown to cause variable effects on the depolymerizing activity of commercial lipases. Pancreatic extracts from both mouse and chicken showed similar depolymerizing activity as the commercial lipases on the P(3HB-co-4HB) film. The presence of lipolytic enzymes in the organ extracts was confirmed with another lipase activity assay, p-nitrophenyl laurate assay. For the first time this has produced a direct evidence for the involvement of lipase-like enzymes from animal in the degradation of this PHA. Lipase is most likely the enzyme from pancreas that was involved in the degradation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0230-z) contains supplementary material, which is available to authorized users.

Nanoparticle and Non-legislated Gaseous Emissions from a Gasoline Direct-Injection Car with Ethanol Blend Fuels and Detergent Additives

Ethanol as an engine fuel emits less pollutants than gasoline, is a completely renewable product with good ecological implications, and has the potential to reduce greenhouse gas emissions. However, ethanol–gasoline blends present a multitude of technical challenges to engine operation, including the creation of adverse engine deposits, especially on inlet valves and injector tips. Special detergent–dispersant additives in ethanol–gasoline blends are the most effective way of bypassing these technical challenges. In the present work, two detergent–dispersant additives, developed by the Oil and Gas Institute, National Research Institute, were used and investigated on a flex-fuel direct-injection vehicle with ethanol–gasoline blend fuels containing 10 and 85% ethanol with special consideration of nanoparticle and non-legislated gaseous emissions. These researchers, among others, stated that, at cold start, particle count emissions are significantly reduced with ethanol blend fuels under all operating condit...

Absorbance and redox based approaches for measuring free heme and free hemoglobin in biological matrices

Cell-free heme (CFH) and hemoglobin (Hb) have emerged as distinct mediators of acute injury characterized by inflammation and microcirculatory dysfunction in hemolytic conditions and critical illness. Several reports have shown changes in Hb and CFH in specific pathophysiological settings. Using PBS, plasma from patients with sickle cell disease, acute respiratory distress syndrome (ARDS) patients and supernatants from red cells units, we found that commonly used assays and commercially available kits do not distinguish between CFH and Hb. Furthermore, they suffer from a variety of false-positive interferences and limitations (including from bilirubin) that lead to either over- or underestimation of CFH and/or Hb. Moreover, commonly used protocols to separate CFH and Hb based on molecular weight (MWt) are inefficient due to CFH hydrophobicity. In this study, we developed and validated a new approach based on absorbance spectrum deconvolution with least square fitting analyses that overcomes these limitations and simultaneously measures CFH and Hb in simple aqueous buffers, plasma or when associated with red cell derived microvesicles. We show how incorporating other plasma factors that absorb light over the visible wavelength range (specifically bilirubin), coupled with truncating the wavelength range analyzed, or addition of mild detergent significantly improves fits allowing measurement of oxyHb, CFH and metHb with >90% accuracy. When this approach was applied to samples from SCD patients, we observed that CFH levels are higher than previously reported and of similar magnitude to Hb.

Optimized Standard Operating Procedures for the Analysis of Cerebrospinal Fluid Aβ42 and the Ratios of Aβ Isoforms Using Low Protein Binding Tubes.

Background: Reduced cerebrospinal fluid (CSF) concentration of amyloid-β1-42 (Aβ1-42) reflects the presence of amyloidopathy in brains of subjects with Alzheimer’s disease (AD).Objective: To qualify the use of Aβ1-42/Aβ1-40 for improvement of standard operating procedures (SOP) for measurement of CSF Aβ with a focus on CSF collection, storage, and analysis.Methods: Euroimmun ELISAs for CSF Aβ isoforms were used to set up a SOP with respect to recipient properties (low binding, polypropylene), volume of tubes, freeze/thaw cycles, addition of detergents (Triton X-100, Tween-20) in collection or storage tubes or during CSF analysis. Data were analyzed with linear repeated measures and mixed effects models.Results: Optimization of CSF analysis included a pre-wash of recipients (e.g., tubes, 96-well plates) before sample analysis. Using the Aβ1-42/Aβ1-40 ratio, in contrast to Aβ1-42, eliminated effects of tube type, additional freeze/thaw cycles, or effect of CSF volumes for polypropylene storage tubes. ‘Low binding’ tubes reduced the loss of Aβ when aliquoting CSF or in function of additional freeze/thaw cycles. Addition of detergent in CSF collection tubes resulted in an almost complete absence of variation in function of collection procedures, but affected the concentration of Aβ isoforms in the immunoassay.Conclusion: The ratio of Aβ1-42/Aβ1-40 is a more robust biomarker than Aβ1-42 toward (pre-) analytical interfering factors. Further, ‘low binding’ recipients and addition of detergent in collection tubes are able to remove effects of SOP-related confounding factors. Integration of the Aβ1-42/Aβ1-40 ratio and ‘low-binding tubes’ into guidance criteria may speed up worldwide standardization of CSF biomarker analysis.

Investigation of the durability and stability of piezoelectric textile fibres

Polymers such as polyvinylidene difluoride, polypropylene and polyamide-11 show great promise for providing light-weight, flexible and fibrous piezoelectric materials that can be integrated into technical textile fabric structures for energy harvesting applications. Durability is an important parameter for the textiles and especially for functional and smart materials. This research work provides an insight on the piezoelectric behaviour of polypropylene, polyamide-11 and polyvinylidene difluoride in terms of peak-to-peak voltage generation capabilities after washing at 40°C with the addition of detergent as described in test method BS EN ISO 105-C06:2010. It was observed that the peak-to-peak voltage generated by polypropylene monofilaments retained similar values with only slight differences, while the monofilaments of polyvinylidene difluoride and polyamide-11 showed higher peak-to-peak voltage generation after washing. These changes have been explained using the changes in the crystallinity and phase, as determined by Fourier transform infrared spectroscopy analysis.

PAH Accessibility in Particulate Matter from Road-Impacted Environments.

Snowmelt, surface runoff, or stormwater releases in urban environments can result in significant discharges of particulate matter-bound polycyclic aromatic hydrocarbons (PAHs) into aquatic environments. Recently, more-specific activities such as road-tunnel washing have been identified as contributing to contaminant load to surface waters. However, knowledge of PAH accessibility in particulate matter (PM) of urban origin that may ultimately be released into urban surface waters is limited. In the present study, we evaluated the accessibility of PAHs associated with seven distinct (suspended) particulate matter samples collected from different urban sources. Laboratory-based infinite sink extractions with silicone rubber (SR) as the extractor phase demonstrated a similar pattern of PAH accessibility for most PM samples. Substantially higher accessible fractions were observed for the less-hydrophobic PAHs (between 40 and 80% of total concentrations) compared with those measured for the most-hydrophobic PAHs (<5% of total concentrations). When we focused on PAHs bound to PM from tunnel-wash waters, first-order desorption rates for PAHs with log Kow > 5.5 were found in line with those commonly found for slowly or very slowly desorbing sediment-associated contaminants. PAHs with log Kow < 5.5 were found at higher desorbing rates. The addition of detergents did not influence the extractability of lighter PAHs but increased desorption rates for the heavier PAHs, potentially contributing to increases in the toxicity of tunnel-wash waters when surfactants are used. The implications of total and accessible PAH concentrations measured in our urban PM samples are discussed in a context of management of PAH and PM emission to the surrounding aquatic environment. Although we only fully assessed PAHs in this work, further study should consider other contaminants such as OPAHs, which were also detected in all PM samples.

Laundry Performance: Effect of Detergent and Additives on Consumer Satisfaction

Abstract To identify key features in the perception of consumer satisfaction of washing performance, a survey among detergent users from five European countries was run. Respondents from each country were volunteers recruited through the Internet without previous selection criteria. The size of the sample (over 4,000 participants) and the wide geographical distribution of the respondents delivered a large set of data. According to the collected answers, respondents show a significant degree of satisfaction with the detergents they use, regardless of type and country. They were only dissatisfied by tough stains and exceptional problems with their wash loads. Detergent additives are widely used but the use and type of laundry additive showed significant differences from country to country. Respondents understood the usefulness of stain removers and showed a high degree of satisfaction using them. Damages to textiles, when it occurs, are not usually associated with the quality of the detergent used but with other factors in the washing process.

Expression of a GDSL esterase from Pseudomonas sp. S9 in Pichia pastoris

Cold active lipolytic enzymes are promising to replace the conventional enzymes processes of biotechnological industries. One of the most important feature of the cold-active lipases and esterases is that they offer economic benefits through energy saving. In general, they exhibit high activity at low temperatures and low thermostability at moderate temperatures. Lipolytic enzyme EstS9 was isolated from Pseudomonas sp. S9. A multiple sequence alignment identified EstS9 as belonging to clade II of the GDSL esterase family. The nucleotide sequence of the estS9 gene of Pseudomonas sp. S9 consists of 1,911 bp. The gene encodes for a protein of 636 amino acid residues with a molecular mass of ∼68.0 kDa. The enzyme is tolerant to alkaline pH and effective at medium to low temperatures (40–25 °C). In our previous study [1], the gene estS9 was expressed using pBAD system in E. coli TOP10 cells as His-tagged fusion protein. Like many other recombinant proteins, EstS9 was produced in E. coli in inclusion bodies. For these reason, there is interest in obtaining the extracellular active esterase which can be used for number of industrial application such as additives for laundry detergents. Therefore, during this study we constructed several recombinant P. pastoris designed for this purpose. Currently, these strains are tested for their ability to produce to the culture medium the active and soluble EstS9 esterase.