Influence of laundry detergents on the abundance dynamics and physiological state of the benthic microalga <Emphasis Type="Italic">Attheya ussurensis</Emphasis> (Bacillariophyta) in laboratory culture

This study examines the influence of the detergents “Obychnyi poroshok” and “Ariel” (at 0.1, 1, and 10 mg/l) on the growth and physiological state of the benthic marine microalga Attheya ussurensis. Cell number, growth rate, and oxygen productivity turned out to be the most sensitive characteristics for evaluation of toxic effects, while the pH of the culture medium and the contents of chlorophyll a and carotenoids were more tolerant to detergent. Toxicants in concentrations of 0.1 and 1 mg/l had a weak inhibitory effect on the microalga A. ussurensis; at 10 mg/l, the detergent Ariel exerted a greater inhibitory effect on the microalga than Obychnyi poroshok did.     

Acceleration of phospholipid flip-flop in the erythrocyte membrane by detergents differing in polar head group and alkyl chain length

The detergents, alkyltrimethylammonium bromide, N-alkyl-N, N-dimethyl-3-ammonio-1-propanesulfonate (zwittergent), alkane sulfonate, alkylsulfate, alkyl-beta-D-glucopyranoside, alkyl-beta-D-maltoside, dodecanoyl-N-methylglucamide, polyethylene glycol monoalkyl ether and Triton X-100, all produce a concentration-dependent acceleration of the slow passive transbilayer movement of NBD-labeled phosphatidylcholine in the human erythrocyte membrane. Above a threshold concentration, which was well below the CMC and characteristic for each detergent, the flip rate increases exponentially upon an increase of the detergent concentration in the medium. The detergent-induced flip correlates with reported membrane-expanding effects of the detergents at antihemolytic concentrations. From the dependence of the detergent concentration required for a defined flip acceleration on the estimated membrane volume, membrane/water partition coefficients for the detergents could be determined and effective detergent concentrations in the membrane calculated. The effective membrane concentrations are similar for most types of detergents but are 10-fold lower for octaethylene glycol monoalkyl ether and Triton X-100. The effectiveness of a given type of detergent is rather independent of its alkyl chain length. Since detergents do not reduce the high temperature dependence of the flip process the detergent-induced flip is proposed to be due to an enhanced probability of formation of transient hydrophobic structural defects in the membrane barrier which may result from perturbation of the interfacial region of the bilayer by inserted detergent molecules.     

Effect of Hard Detergents on Algae in a High-Rate-Oxidation Pond

Regular concentrations of hard detergents in domestic wastewater do not affect algal growth in a high-rate-oxidation pond. The addition of nonionic hard detergent at concentrations above 60 mg/liter decreased the algal concentration in the batch culture, and complete lysis of algal cells was observed within a few days at a detergent concentration of 100 mg/liter.     

Toxic effects of detergents on the alga Plagioselmis prolonga (Cryptophyta)

The effect of sodium dodecyl sulfate (SDS) and the household synthetic detergents (HSDs) Kristall and Tix (0.1, 1, and 10 mg/l) on cell motility, cell number dynamics, and the growth rate of the alga Plagioselmis prolonga (Cryptophyta) is studied. Algal cell motility proved to be the most sensitive indicator of detergent toxicity. SDS was the least toxic: 1 mg SDS/l caused a short-term loss of motility in 10% of the algal cells. The HSD Tix was the most toxic: only 70% of the cells recovered motility after a 24 h exposure to 1 mg/l. The substances tested in a concentration of 10 mg/l caused mortality of the P. prolonga population. According to their toxic effect on P. prolonga, the investigated toxicants can be arranged as follows: SDS < Kristall < Tix.     

Influence of the Linear Alkylbenzene Sulfonate (LAS) on hematological and biochemical parameters of Nile Tilapia,Oreochromis niloticus

The acute toxicity of household detergent (Ariel) on blood parameters and histology of Oreochromis niloticus was investigated using static bioassay for 96 h. Linear alkylbenzene sulfonate (LAS) is an anionic surfactant widely used in detergents and cleaners, both in industrial and household applications. LAS contaminating aquatic ecosystems as a potential toxic pollutant, was investigated in the present study for acute toxicity. The fish samples were divided into six groups, including 20 fish in each group. Normal feed was given to control group without detergents treatment. Hematological parameters (RBC count, Hb, Ht and platelets) were significantly declined, while WBC count showed a highly significant increase. Compared with the control group, significant elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was recorded in fish treated with different concentrations of detergent. Catalase (CAT), Superoxide dismutase (SOD) activities and Reduced Glutathione (GSH) concentration showed a highly significant reduction. Total proteins showed significant decrease, while total lipids, cholesterol and triglycerides significantly increased. The mean lethal concentration (LC₅₀) for 96 h of Ariel was at concentration 10 mg/L. Relative percentage of detergent residues in fish muscles was increased with higher detergent concentrations. In conclusion, exposure to detergents resulted in great alterations in the histological structure of liver and gills.     

家用洗涤剂对红剑鱼、孔雀鱼和食蚊鱼的急性毒性实验观察

目的 研究4种不同产地的家用洗涤剂对红剑鱼、孔雀鱼和食蚊鱼的急性毒性影响。方法 用水毒理学方法测定它们对3种鱼类的24hLC50,并将洗涤剂溶液存放15d和30d后测定其对鱼类的急性毒性。结果 在远低于日常使用量的浓度下,4种家用洗涤剂均对红剑鱼、孔雀鱼和食蚊鱼有急性毒性,24h LC50值在20mg／L～55mg／L;家用洗涤剂溶液存放一段时间后对鱼类的急性毒性作用无明显降低。结论 含有大量家用洗涤剂的生活污水排放到自然水体后将对鱼类产生持续的有害影响。     

Effect of chemical water pollution on the physiological activity of microorganisms

Anionactive detergents occurring in water in concentrations up to 100 micrograms . ml-1 do not exert inhibitory effects on the present microflora, either in case of heterotrophic bacteria, indicators of fecal pollution, or pathogenic and facultative pathogenic bacteria, including viruses. On the contrary, the studied bacterial species are able to decompose surfactants and utilize them as carbon source. Though, the rate of degradation is different with respect to respective substances, as well as with regard to the species of organisms, and their physiological activity. Only at higher detergent concentrations inhibition of microbial cell growth may be observed.     

Detergent effects on enzyme activity and solubilization of lipid bilayer membranes

Over 50 detergents were tested to establish which would be most effective in releasing proteins from membrane-bounded compartments without denaturating them. Various concentrations of each detergent were tested for two activities: (1) solubilization of egg phospholipid liposomes as measured by reduction of turbidity and (2) effect of detergent concentration on the activities of soluble, hydrolytic enzymes. Those detergents must effective in solubilizing 0.2% lipid and least detrimental to enzymes were five pure, synthetic compounds recently introduced: CHAPS, CHAPSO, Zwittergents 310 and 312, and octylglucoside. Industrial detergents were generally much inferior, insofar as they solubilized membranes inefficiently and/or inactivated certain hydrolytic enzymes readily. The five detergents were characterized by (a) an unusually high critical micelle concentration and (b) a preference for forming mixed micelles with lipids instead of forming pure micelles, as indicated by an ability to solubilize lipid at concentrations of detergent significantly below the critical micelle concentration. This characteristic permits solubilization of high concentrations of membrane below the critical micelle concentration of the detergent so that protein denaturation is minimized. A generally applicable guideline that emerged from this study is that detergents should be used at approximately their critical micelle concentration which should not be exceeded by the concentration of membrane. Similar considerations should apply to the use of detergents in purifying and reconstituting intrinsic membrane proteins.     

Effect of detergents on the association of the glycophorin a transmembrane helix

We have examined the role of the environment on the interactions between transmembrane helices using, as a model system, the dimerization of the glycophorin A transmembrane helix. In this study we have focused on micellar environments and have examined a series of detergents that include a range of alkyl chain lengths, combined with ionic, zwitterionic, and nonionic headgroups. For each we have measured how the apparent equilibrium constant depends on the detergent concentration. In two detergents we also measured the thermal sensitivity of the equilibrium constant, from which we derive the van't Hoff enthalpy and entropy. We show that several simple models are inadequate for explaining our results; however, models that include the effect of detergent concentration on detergent binding are able to account for our measurements. Our analysis suggests that the effects of detergents on helix association are due to a pair of opposing effects: an enthalpic effect, which drives association as the detergent concentration is increased and which is sensitive to the chemical nature of the detergent headgroup, opposed by an entropic effect, which drives peptide dissociation as the detergent concentration is raised. Our results also indicate that the monomer-monomer interface is relatively hydrophilic and that association within detergent micelles is driven by the enthalpy change. The wide variations in glycophorin a dimmer, stability with the detergent used, together with the realization that this results from the balance between two opposing effects, suggests that detergents might be selected that drive association rather than dissociation of peptide dimers.     

Solubilization of UDP-glucose collagen glucosyltransferase activities from chick embryo liver: Golgi apparatus, smooth and rough endoplasmic reticulum.

1. The choice of a suitable detergent for solubilization of UDP-glucose collagen glucosyltransferase (GGT) activities from chick embryo liver has been investigated. Several detergents were used (zwitterionic detergent as Chaps, and non-ionic detergents as Triton X-100, Nonidet P 40, Brij 35). 2. All the detergents with GGT activities were tested in Golgi apparatus, smooth and rough endoplasmic reticulum (SER, RER). 3. 80-100% GGT Golgi apparatus activity was easily solubilized at low concentrations in surfactant (0.5 mg/ml). 25-78% of SER and RER GGT activities were extracted at this concentration. 4. A higher level of detergent (5 mg/ml) was necessary to release all GGT activities of SER and RER. Protein extraction was identical to GGT activities.     

Mortality patterns in trout (Salmo trutta and S. gairdneri) exposed to an anionic detergent in relation to concentration and mechanisms of toxic action

SUMMARY. Histological and electron microscopical study of the gills of fish poisoned by the anionic detergent sodium lauryl sulphate indicates that the nature of the toxic action changes at a concentration of about 120 mg I⁻¹. Toxicity tests were conducted to determine whether this change could be detected in the test data, using brown trout ( Salmo trutta ) and rainbow trout ( S. gairdneri ).
Toxicity curves showed no indication of the change in toxic action, but at concentrations of 120 and 130 mg I⁻¹ of sodium lauryl sulphate 'split' probit lines occurred, and the lines for 150 mg I⁻¹ consistently differed in slope from other lines.     

Toxic action of several lethal concentrations of an anionic detergent on the gills of the brown trout (Salmo trutta L.)

The pathological effects often lethal concentrations of the anionic detergent, sodium lauryl sulphate, on the gills of Salmo trutta L. have been studied by light and electron microscopy. At concentrations to 120 mg/1 (medium survival times >1 h), epithelial cell death is associated with lysosome formation. Acute inflammation of the gill tissue, extensive detachment of the epithelium and, except at the lowest concentrations, collapse of the pillar cell system occur. At concentrations above 120 mg/1 (medium survival times <1 h) very rapid lysis of cells results in the complete disruption of cellular and tissue structure. Changes in the gross structure of the gills are explainable in terms of the rate and nature of toxic action at the cellular level. Review of the biomedical literature suggests the observed effects of sodium lauryl sulphate on gill cells correspond to the two mechanisms by which detergents cause death in isolated cells. These are autolysis, i.e. lysis by the action of the cell's own enzymes, induced by an initial lesion in the cell membrane whose precise nature is not known; and rapid lysis by the direct action of the detergent on the cell constituents.     

Detergent-assisted refolding of guanidinium chloride-denatured rhodanese. The effect of lauryl maltoside

For the first time, the enzyme rhodanese (thiosulfate:cyanide sulfurtransferase; EC 2.8.1.1) has been renatured from 6 M guanidinium chloride (GdmCl) by direct dilution of the denaturant at relatively high protein concentrations. This has been made possible by using the nonionic detergent dodecyl-beta-D-maltoside (lauryl maltoside). Lauryl maltoside concentration dependence of the renaturation and reactivation time courses were studied using 50 micrograms/ml rhodanese. There was no renaturation at lauryl maltoside (less than 0.1 mg/ml), and the renaturability increased, apparently cooperatively, up to 5 mg/ml detergent. This may reflect weak binding of lauryl maltoside to intermediate rhodanese conformers. The renaturability began to decrease above 5 mg/ml lauryl maltoside and was significantly reduced at 20 mg/ml. Individual progress curves of product formation, for rhodanese diluted into lauryl maltoside 90 min before assay, showed induction phases as long as 7 min before an apparently linear steady state. The induction phase increased with lauryl maltoside concentration and could even be observed in native controls above 1 mg/ml detergent. These results are consistent with suggestions that refolding of GdmCl-denatured rhodanese involves an intermediate with exposed hydrophobic surfaces that can partition into active and inactive species. Further, lauryl maltoside can stabilize those surfaces and prevent aggregation and other hydrophobic interaction-dependent events that reduce the yield of active protein. The rhodanese-lauryl maltoside complex could also form with native enzyme, thus explaining the induction phase with this species. Finally, it is suggested that renaturation of many proteins might be assisted by lauryl maltoside or other "nondenaturing" detergents.     

A novel prokaryotic expression system for biosynthesis of recombinant human membrane-bound catechol-O-methyltransferase

Membrane proteins constitute 20-30% of all proteins encoded by the genome of various organisms. While large amounts of purified proteins are required for pharmaceutical and crystallization attempts, there is an unmet need for the development of novel heterologous membrane protein overexpression systems. Specifically, we tested the application of Brevibacillus choshinensis cells for the biosynthesis of human membrane bound catechol-O-methyltransferase (hMBCOMT). In terms of the upstream stage moderate to high expression was obtained for complex media formulation with a value near 45 nmol/h/mg for hMBCOMT specific activity achieved at 20 h culture with 37 °C and 250 rpm. Subsequently, the efficiency for reconstitution of hMBCOMT is markedly null in the presence of ionic detergents, such as sodium dodecyl sulphate (SDS). In general, for non-ionic and zwiterionic detergents, until a detergent critic micellar concentration (CMC) of 1.0 mM, hMBCOMT shows more biological activity at lower detergent concentrations while for detergent CMC higher than 1 mM, higher detergent concentrations seem to be ideal for hMBCOMT solubilization. Indeed, from the detergents tested, the non-ionic digitonin at 0.5% (w/v) appears to be the most suitable for hMBCOMT solubilization.     

Utilization of zeolite Y in the removal of anionic,cationic and nonionic detergents during purification of proteins

Summary Hydrophobie zeolite Y was used to adsorb detergents from protein solutions and within one minute the commonly used detergents sodium dodecyl sulfate, cetyl trimethyl ammonium bromide, and Triton X-100 at concentrations of 10 mg/ml were adsorbed to a level below their critical micelle concentrations. From the detergent depleted solutions 77 to 85 % of the proteins were recovered; the lower value was obtained with protein concentration below one mg/ml.     

Effects of chaotropic agents versus detergents on dihydroorotate dehydrogenase.

As chaotropic salts are generally believed to affect water structure in a manner which increases lipophilicity of water, they may seem to be capable of substituting for detergents in the solubilization of particulate enzyme. Although solubilization either by detergents or by chaotropic salts has been demonstrated with several membrane proteins, the effects these agents have on the properties and activity of an enzyme may be quite different. This is illustrated by the effects on mammalian mitochondrial dihydroorotate dehydrogenase. Stability of the solubilized enzymic activity is dependnet on the presence of a detergent and maximum enzymic activity is observed at the critical micelle concentration of the detergent. Addition of low concentrations of various anions of the chaotropic series further enhances activity while higher concentrations of these anions, although increasing solubility of the enzyme, irreversibly inhibit catalysis.     

Solubilization of Myelin Membranes by Detergents

The major proteins of myelin have classically been extracted in organic solvents. Here we investigated some of the characteristics of brain myelin solubilization in aqueous detergent solutions. At comparable molar concentrations, two nonionic detergents, i.e., octyl glucoside and Lubrol PX, proved relatively better myelin solubilizers than the detergents related to the bile salts, i.e., cholate and CHAPS. The two former detergents solubilized more protein than lipid and the two latter ones more lipid than protein from myelin membranes. All four detergents solubilized the phospholipid more efficiently than the cholesterol component of myelin. The detergent concentrations required for myelin solubilization were reduced substantially if the temperature and the salt concentration of the media were increased. As much as 3 mg of lyophilized myelin (about 1 mg of protein) were solubilized readily per milliliter of a solution containing 30 mM octyl glucoside and 0.1 M sodium sulfate in 0.1 M sodium phosphate buffer, pH 6.7. Each of the detergents studied, including the above four, sodium dodecyl sulfate (SDS). Triton X-100, and Zwittergent 3-14, had its own advantages and drawbacks as myelin protein extractors. The nonionic amphiphiles and CHAPS left a small residue mainly composed of proteins of the Wolfgram fraction, as revealed by SDS-polyacrylamide gel electrophoresis. Octyl glucoside was preferred, given its versatility as solubilizer, ultraviolet transparency, and high critical micellar concentration. Observations on possible difficulties that may be encountered are also included.     

Detergents as probes of reconstituted rat liver cytochrome P-450 function

A series of 16 ionic, zwitterionic, and nonionic detergents have been used to perturb the catalytic activities of major cytochrome P-450 (P-450) forms from untreated (UT-A), phenobarbital-treated (PB-B) and beta-naphthoflavone-treated (BNF-B) rats in reconstituted systems with NADPH--P-450 reductase Detergent effects on R warfarin hydroxylase activities were correlated with detergent effects on the quaternary structures of P-450 and reductase, and on their 1:1 complexes as determined by gel exclusion chromatography using sodium cholate as a prototype detergent. The detergent concentrations used did not in most cases affect rates of NADPH-dependent reduction of cytochrome c by the reductase. With P-450 BNF-B, ionic and zwitterionic detergents enhanced warfarin hydroxylase activities at low concentrations and produced marked inhibition at higher concentrations, while nonionic detergents only inhibited. With P-450 UT-A, some nonionic and zwitterionic detergents increased rates at low concentrations and inhibited at higher concentrations. P-450 PB-B was inhibited by detergents of all three classes at low and high concentrations. The concentrations of a detergent required to affect 50% inhibition differed for the three P-450s, suggesting, together with the differential susceptibilities to detergent-mediated rate enhancing effects, that the reductase interacts functionally differently with the three P-450s. Chromatographic studies demonstrated that concentrations of sodium cholate which optimally enhanced metabolic rates with P-450 BNF-B facilitated the uptake of the P-450 into the functional reductase/P-450 complex, and higher concentrations of cholate, which completely inhibited activity, produced profound disruptions of the complex. The data have provided insight into the functional interactions required for monooxygenase activity.     

Activation, inhibition, and destabilization of Thermomyces lanuginosus lipase by detergents

Lipases catalyze the hydrolysis of triglycerides and are activated at the water-lipid interface. Thus, their interaction with amphiphiles such as detergents is relevant for an understanding of their enzymatic mechanism. In this study, we have characterized the effect of nonionic, anionic, cationic, and zwitterionic detergents on the enzymatic activity and thermal stability of Thermomyces lanuginosus lipase (TlL). For all detergents, low concentrations enhance the activity of TlL toward p-nitrophenyl butyrate by more than an order of magnitude; at higher detergent concentrations, the activity declines, leveling off close to the value measured in the absence of detergent. Surprisingly, these phenomena mainly involve monomeric detergent, as activation and inhibition occur well below the cmc for the nonionic and zwitterionic detergents. For anionic and cationic detergents, activation straddles the monomer-micelle transition. The data can be fitted to a three state interaction model, comprising free TlL in the absence of detergent, an activated complex with TlL at low detergent concentrations, and an enzyme-inhibiting complex at higher concentrations. For detergents with the same headgroup, there is an excellent correspondence between carbon chain length and ability to activate and inhibit TlL. However, the headgroup and number of chains also modulate these effects, dividing the detergents overall into three broad groups with rising activation and inhibition ability, namely, anionic and cationic detergents, nonionic and single-chain zwitterionic detergents, and double-chain zwitterionic detergents. As expected, only anionic and cationic detergents lead to a significant decrease in lipase thermal stability. Since nonionic detergents activate TlL without destabilizing the protein, activation/inhibition and destabilization must be independent processes. We conclude that lipase-detergent interactions occur at many independent levels and are governed by a combination of general and structurally specific interactions. Furthermore, activation of TlL by detergents apparently does not involve the classical interfacial activation phenomenon as monomeric detergent molecules are in most cases responsible for the observed increase in activity.     

Detergent quantification in membrane protein samples and its application to crystallization experiments

The structural characterization of membrane proteins remains a challenging field, largely because the use of stabilizing detergents is required. Researchers must first select a suitable detergent for the solubility and stability of their protein during in vitro studies. In addition, an appropriate concentration of detergent in membrane protein samples can be essential for protein solubility, stability, and experimental success. For example, in membrane protein crystallography, detergent concentration in the crystallization drop can be a critical parameter influencing crystal growth. Over the past decade, multiple techniques have been developed for the measurement of detergent concentration using a wide variety of strategies. These methods include colorimetric reactions, which target specific detergent classes, and analytical techniques applicable to a wide variety of detergents. This review will summarize and discuss the available options. It will be a useful resource to those selecting a strategy that best fits their experimental requirements and available instruments.