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Pozyskiwanie bakterii wykazujących zdolność dekoloryzacji barwników azowych

Zabłocka-Godlewska Ewa, Przystaś Wioletta

Proceedings of ECOpole

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2020

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Vol. 14, No. 1

141--153

PL

Barwniki azowe to najliczniejsza i najczęściej stosowana grupa barwników syntetycznych. W większości są to substancje odporne na działanie czynników fizycznych, chemicznych oraz biologicznych, co wpływa na trudności w efektywnym ich usuwaniu ze ścieków i stwarza ryzyko ich przenikania do środowiska wodnego. Badania nad zwiększeniem sprawności konwencjonalnych metod oczyszczania ścieków barwnych obejmują m.in. pozyskiwanie mikroorganizmów o wysokim potencjale dekoloryzacyjnym, które można będzie wykorzystać np. do bioaugmentacji układów biologicznego oczyszczania. Celem badań był screening bakterii wykazujących zdolność dekoloryzacji dwóch barwników azowych (błękitu Evansa (BE), czerwieni Kongo (CK)). Mikroorganizmy izolowano ze ścieków komunalnych, kompostu oraz spróchniałego drewna bukowego. W pierwszym etapie izolacji wykorzystano podłoża stałe (mineralne i agar odżywczy) suplementowane barwnikami należącymi do dwóch grup chemicznych (disazowy błękit Evansa (BE), trifenylometanowa zieleń brylantowa (ZB)). Izolaty kilkukrotnie pasażowano w celu potwierdzenia ich czystości i potencjału dekoloryzacyjnego. W drugim etapie badań zastosowano hodowle płynne na podłożu bulion odżywczy zawierającym barwniki syntetyczne (BE lub CK) w stężeniu 0,1 g/dm3. Pomiary spektrofotometryczne prób po dekoloryzacji posłużyły do określenia zawartości barwników i obliczenia ich procentowego usunięcia. Wykazano, że wszystkie wykorzystane materiały mogą stanowić źródło pozyskiwania bakterii dekoloryzujących. Największą ich liczbę izolowano ze ścieków komunalnych, następnie kompostu i próchna. Bardziej skomplikowany strukturalnie BE był efektywniej usuwany przez bakterie niż CK. Większą efektywność dekoloryzacji wykazywały szczepy izolowane na podłożu mineralnym. Spośród nich 3 izolowane na podłożu MM charakteryzowały się szczególnymi właściwościami, usuwały zadane barwniki już po 48 h hodowli z efektywnością przekraczającą 90 %.

EN

Azo dyes are the most numerous and most commonly used group of synthetic dyes. Most of them are resistant to physical, chemical and biological factors. It causes difficulties in effective removal of dyes from wastewater and creates the risk of their penetration into the aquatic environment. Research on increasing the efficiency of conventional methods of treatment of the coloured wastewater includes, inter alia, obtaining microorganisms with high decolourization potential. Such microorganisms can be used e.g. for bioaugmentation of biological treatment systems. The aim of the study was screening of bacteria showing the ability to decolorize two azo dyes (Evans blue (EB), Congo red (CR). Microorganisms were isolated from municipal sewage, compost and rotten beech wood. At the first stage of screening were used two kinds of solid growth media (mineral and nutrient agar) supplemented with dyes belonging to two chemical groups (disazo Evans blue (EB); triphenylmethane brilliant green (BG)). Isolates were passaged several times to confirm their purity and decolourization potential. In the second stage of research, liquid cultures on nutrient broth containing synthetic dyes (EB or CR) at a concentration of 0.1 g /dm3 were used. The content of dyes in samples after decolourization were measure spectrophotometrically and their percentage removal was calculated. It has been shown that all used materials can be a source for obtaining decolorizing bacteria. The largest number of them was isolated from municipal wastewater, followed by compost and rotten wood. Structurally more complicated EB was more efficiently removed by bacteria than CR. Strains isolated on mineral media showed greater decolourization efficiency. Amongst the strains isolated on MM, 3 of them removed dyes already after 48 h of culture with an efficiency exceeding 90 %.

2

Radiation induced degradation of Congo red dye: a mechanistic study

Muner Majid, Saeed Muhammad, Bhatti Ijaz Ahmad, Haq Atta-ul, Khosa Muhammad Kaleem, Jamal Muhammad Asghar, Ali Saddaqat

Nukleonika

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2019

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Vol. 64, No. 2

49--53

EN

Synthetic dyes are persistent pollutants with poor biodegradability. The present study is about the degradation of direct Congo red dye in aqueous media using the Co-60 gamma radiation source. The experimental conditions such as gamma-ray absorbed doses, amount of oxidant (H2O2) and pH conditions were evaluated. The lambda max of dye solution was noted as 498 nm, and then, decrease in absorbance and reduction in chemical oxygen demand (COD) were examined. The complete colour removal of dye was observed at 5 kGy, while a signifi cant COD removal was observed at 15 kGy gamma-ray absorbed dose in conjunction with oxidant for 50 mg/L concentration. It was found that pH has no influence on degradation efficiency. A possible degradation pathway was proposed. The radiolytic end products were monitored by Fourier transform infrared (FTIR) and gas chromatography coupled with mass spectrometry (GC-MS) to explore the degradation mechanism. It was imperative to study the oxidative degradation pathway to provide directions for potential applicability of advanced oxidation process (AOP) in industrial wastewater treatment.

3

Adsorption of congo red from aqueous solutions by porous soybean curd xerogels

Zhang Z., Li Y., Du Q., Li Q.

Polish Journal of Chemical Technology

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2018

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Vol. 20, nr 3

95--102

EN

Soybean curd is a very popular food containing high-quality protein, polyunsaturated fats, vitamins, minerals and other nutrients. This study aims to prepare porous soybean curd xerogels via a vacuum freeze drying method and uses them as adsorbents to remove congo red from aqueous solutions. The morphology and functional groups of the soybean curd xerogels were characterized using scanning electron microscopy and Fourier transform infrared spectroscopy, respectively. The adsorption properties of congo red onto the soybean curd xerogels were carried out through investigating the influencing experimental parameters such as the drying method, solution pH, adsorbent dose, contact time and temperature. The results showed that the adsorption isotherm data were fitted well to the Freundlich isotherm. Adsorption kinetics of congo red onto the soybean curd followed the pseudo-second-order kinetic model. The thermodynamic parameters, such as ΔG°, ΔH° and ΔS°, were also determined.

4

Congo red fluorescence upon binding to macromolecules – a possible explanation for the enhanced intensity

Zemanek G., Jagusiak A., Chłopaś K., Piekarska B.

Bio-Algorithms and Med-Systems

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2017

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Vol. 13, no. 2

69--78

EN

The present study attempts to explain the reason for the selective generation of an increase in intensity of Congo red (CR) fluorescence as an effect of the dye interacting with proteins and polysaccharides. This supramolecular dye, which creates ribbon-shaped micelles in aqueous solutions when excited with blue light (470 nm), presents low fluorescence with a maximum within the orange-red light range (approximately 600 nm). In the same conditions, CR-stained preparations of heat-denatured proteins, some native proteins (e.g. cell surface receptors) and cellulose show intense orange-red fluorescence when observed using a fluorescence microscope. The fluormetric measurements showed that the factors that cause the dissociation of the ribbon-shaped CR micelle – ethanol, urea, dimethyl sulfoxide (DMSO) and cholate – all contributed to a significant increase in the fluorescence intensity of the CR solutions. The fluorescence measurements of CR bound to the immunoglobulin light lambda (L λ) chain and soluble carboxymethyl cellulose (CMC) showed a fluorescence intensity which was many times higher. In the case of the denatured (65°C) immunoglobulin L λ chain, the fluorescence intensity significantly exceeded the values observed for the factors which break down the CR micelles. The dissociation of the ribbon-shaped micelles and the complexation of the monomeric CR form with polymers are two of the factors explaining the intense fluorescence of protein and polysaccharide preparations stained with CR.

5

Shortening and dispersion of single-walled carbon nanotubes upon interaction with mixed supramolecular compounds

Jagusiak A., Piekarska B., Chłopaś K., Bielańska E., Pańczyk T.

Bio-Algorithms and Med-Systems

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2016

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Vol. 12, no. 3

123--132

EN

Congo red (CR) dye molecules self-associate in water solutions creating ribbon-like supramolecular structures that can bind various aromatic compounds by intercalation, forming mixed supramolecular systems. Mixed supramolecular systems, such as CR-doxorubicin and CREvans blue, interact with the surface of carbon nanotubes, leading to their stiffening and ultimately to their breaking and shortening. This work presents a simple method of obtaining short and straight carbon nanotubes with significantly better dispersion in aqueous solutions and consequently improved usability in biological systems.

6

The use of Titan yellow dye as a metal ion binding marker for studies on the formation of specific complexes by supramolecular Congo red

Chłopaś K., Jagusiak A., Konieczny L., Piekarska B., Roterman I., Rybarska J., Stopa B., Zemanek G., Bielańska E., Piwowar P., Sadlik K.

Bio-Algorithms and Med-Systems

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2015

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Vol. 11, no. 1

9--17

EN

Congo red (CR) and other self-assembling compounds creating supramolecular structures of rod- or ribbon-like architecture form specific complexes with cellulose and also with many proteins, including antibodies bound to the antigen and amyloids in particular. The mechanism of complexation and structure of these complexes are still poorly recognized despite the importance of the problem for medicine. This work proposes the progress in electron microscopy studies of amyloid-dye complexes by labeling supramolecular ligand CR with silver ions as a marker. Silver ions are introduced to CR carried by the strongly binding silver dye Titan yellow, which in addition form comicellar structures with CR. Silver carried by self-assembled dye molecules forms in the resulting metal nanoparticles, making the specific amyloid ligand CR perceptible in EM studies.

7

Attempt at a systemic outlook on aging and carcinogenesis

Piwowar M., Dygut J., Piwowar P., Konieczny L., Roterman I.

Bio-Algorithms and Med-Systems

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2014

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Vol. 10, no. 3

101--115

EN

Two of the key problems plaguing humanity – aging and carcinogenesis – are inexorably linked. While their nature seems different, their mechanisms have a lot in common. Evidence suggests that aging is the result of spontaneous synthesis and accumulation of improperly folded proteins in cells, leading to a variety of pathologies. As for carcinogenesis, it is tied to genetic mutations – permanent, covalent changes in the DNA. Both processes are random in character; however, unlike mutations, the accumulation of malformed proteins is not genetically determined. Instead, control over this process hinges upon regulating the protein exchange rate – a phenomenon that seems a likely candidate for the basic aging control mechanism. Although mutations themselves may be counteracted in a controlled manner, their effects typically cannot. The mechanisms of aging and carcinogenesis, while functionally different, remain correlated: an aging cell is rendered more susceptible to mutational changes. The rapidly growing body of information regarding aging and carcinogenesis enables a systemic approach to both these phenomena – an approach that is attempted in this review.

8

Application of numerical analysis of fluorescence spectrum to identify properties of substances associating with Congo red micelle

Zemanek G., Jagusiak A., Kusior D., Piekarska B., Spólnik P., Stopa B.

Bio-Algorithms and Med-Systems

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2011

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Vol. 7, no. 2

17--23

EN

The particles of Congo red (CR), a bis-azo dye, associate in aqueous solutions, forming ribbon-like micelles. Supramolecular dyes, as a result of their molecular size, have a limited capability of penetrating into the interior of the majority of native proteins and do not enter living cells surrounded by an intact phospholipid membrane. They may however, bind to proteins with structure destabilized by unfolding (during denaturation) or function-related changes (e.g. surface cell receptors). Congo red absorbs visible light with a maximum absorption at 500 nm, in aqueous solutions at neutral pH. After CR bind to the polymers (such as cellulose) or certain proteins, dye’s behaviour changes remarkably. These binding affects spectral properties of the dye: the absorption spectrum of the Congo red shifts towards longer wavelengths and receives a yellow-red fluorescence. A numerical analysis of the graphic data obtained from fluorescent microscope (division into channels corresponding to constituent colours and with the noise separated using a graphical program) allowed to understand better the causes of movement of the CR fluorescence spectrum and to identify the properties of substances associating with the CR micelle. By means of a statistical analysis of the data it was possible to observe that wavelength of emission and intensity of fluorescence is not only associated with the polarity of the environment but mainly with the strength of the association between the Congo red micelle and the matrix.

9

The effect of Congo red inhibitor on the corrosion of various steels in a 3.5% NaCl medium

Sahin M., Asan A., Celikkan H., Aksu M. L.

Materials Science Poland

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2010

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Vol. 28, No. 4

795--803

EN

This study is concerned with the use of Congo red as an inhibitor for low alloyed carbon steel, petroleum steel and boron steel at 60 °C in 3.5% NaCl aqueous solution. Analysis was performed using the Tafel polarization measurements and electrochemical impedance spectroscopy. Congo red was observed to cover the surface by adsorbing upon it, and its inhibition efficiency depended on the concentration. The efficiency was the highest one in low-alloyed carbons steels, followed by petroleum and boron steels. The type of adsorption occurring on the metal surface was also determined.

10

The Approach For Computer Analysis Of Results Obtained In Two-Dimensional Electrophoresis Of Serum Protein Complexes With Congo Red

Śmietański J.

Bio-Algorithms and Med-Systems

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2006

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Vol. 2, no. 3

25--31

EN

The research of blood serum by the method of double electrophoretical separation, where the second, perpendicular run is modified by adding Congo red, inspires a lot of hope. This paper describes the attempt to create a program which is indispensable to interpret the results. The received on input scan of the glass plate is brought under preliminary processing which enables to get us rid of noise and useless information. The segmentation which is done in order to separate the objects from the background, was implemented by the method of linear image search and merge. The points are added from the darkest to the brightest ones to the single objects. The recognition of previously separated objects proceeds stage by stage, thanks to that the areas which are difficult to identify can have more distinctive marks. The value of appurtenance function is calculated by the nearest neighbour method, and for the final classification the majorizating rule is used. The calculation of percentage of each fraction proceeds by summing the brightness of the points which make the object, after the value of the background was substracted. The obtained results of the automatic recognition are promising. However, a greater amount of experimental images is needed for the full evaluating of the usefulness of the program.

PL

Badanie składu surowicy krwi metodą podwójnego rozdziału elektroforetycznego, gdzie drugi, prostopadły rozdział modyfikowany jest dodaniem czerwieni Kongo, budzi spore nadzieje. W niniejszej pracy przedstawiam próbę stworzenia oprogramowania niezbędnego do interpretacji wyników. Otrzymany na wejściu skan szklanej płytki poddawany jest wstępnej obróbce, umożliwiającej pozbycie się szumu oraz zbędnych informacji. Segmentacja, mająca na celu wyodrębnienie poszczególnych obiektów od tła, zaimplementowana została metodą liniowego przeglądania obrazu i sklejeń. Punkty do poszczególnych obiektów dodawane są w kolejności od najciemniejszych do najjaśniejszych. Rozpoznanie uprzednio wyodrębnionych obiektów odbywa się etapowo, dzięki czemu trudniej identyfikowalne plamy mają do dyspozycji większą liczbę charakteryzujących je cech. Wartość funkcji przynależności obliczana jest metodą najbliższego sąsiada, a do ostatecznej klasyfikacji stosujemy regułę majoryzacyjną. Obliczenie procentowego udziału każdej frakcji następuje poprzez zsumowanie jasności punktów składających się na obiekt, po uprzednim odjęciu wartości tła. Uzyskane rezultaty automatycznego rozpoznania są obiecujące, jednak do pełnej oceny przydatności programu konieczna jest większa liczba obrazów eksperymentalnych.

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Tworzenie i rozdział kompleksów białek surowicy z barwnikami supramolekularnymi w elektroforezie dwukierunkowej - próba opracowania komputerowej techniki analizy obrazu rozdziałowego

Rybarska J., Spólnik P., Drozd A., Konieczny L., Piekarska B., Stopa B., Zemanek G., Król M., Roterman-Konieczna I.

Bio-Algorithms and Med-Systems

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2005

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Vol. 1, no. 1/2

133--138

PL

Praca przedstawia badania eksperymentalne mające na celu wykrycie zmian w składzie białkowym zmienionej chorobowo surowicy ludzkiej. Dla wykrycia zmian opracowano dwukierunkową technikę elektroforetyczną umożliwiającą tworzenie kompleksów przez barwniki o charakterze ciekłokrystalicznym z białkami. Taśmowe micele barwnika mogą przylegać do szkieletu łańcucha polipeptydowego w obszarach jego ekspozycji, tworząc kompleksy. Warunki elektroforetyczne pozwalają na usunięcie nadmiaru oraz puli słabo związanego z białkami barwnika. Tworzenie kompleksów w elektroforezie pozwala na ujawnienie kompleksów białek z barwnikami, które wędrują szybciej niż białka niezaangażowane w wiązanie barwnika. Nieprawidłowe oraz przejściowo zdestabilizowane natywne białka mogą być podatne na penetrację i wiązanie dużych supramolekularnych ligandów. Do grupy białek wiążących należą białka szpiczakowe oraz serpiny, haptoglobiny i immunoglobuliny, ulegające przegrupowaniu podczas tworzenia kompleksów ze swoimi fizjologicznymi ligandami. Elektroforeza surowicy prowadzona jest dwuetapowo w prostopadłych względem siebie kierunkach. Pierwszy rozdział jest standardowy. Drugi modyfikowany jest przez kontakt i ewentualną interakcję rozdzielonych uprzednio białek z dodanym barwnikiem. Szybciej wędrujący barwnik, którego punkt nałożenia znajduje się poniżej białek, napotyka w trakcie wędrówki i wyprzedza białko. Białka zdolne do tworzenia kompleksów zabarwiają się w takich warunkach oraz wędrują szybciej od swoich nieskompleksowanych z barwnikiem odpowiedników. Po redukcji barwników dwutioninem sodowym z następowym zabarwieniem błękitem bromofenolowym, można uzyskać kompletny proteinogram rozdzielanej surowicy. Próby opracowania pół-automatycznej i automatycznej analizy uzyskiwanego w elektroforezie rozdziału plam zostały ostatnio podjęte.

EN

Two-dimensional agarose electrophoresis was used to create suitable conditions for the formation of protein complexes with supramolecular dyes. Ribbon-shaped dye molecule ligands adhere during complexation to the polypeptide backbone of β-conformation within protein clefts. The electrophoresis helps to remove the dye excess and the dye weakly attached to protein molecules. It simultaneously allows for the exposure of protein-dye complexes migrating faster than proteins not engaged in complexation. Abnormal and transiently destabilized native proteins become susceptible to penetration and binding by large supramolecular dye ligands. This includes many myeloma proteins as well as serpins, haptoglobins and immunoglobulins engaged in their natural complexes. Electrophoresis of serum proteins is conducted in two steps: the first is a standard electrophoresis while the second (perpen-dicular run) is modified by the contact and interaction of separated serum proteins (during the first step) with the added dye. The fast migrating dye, for which the starting position is retreated versus to that of proteins, meets and overruns the proteins. The proteins which are susceptible to dye penetration and binding become stained and their migration is accelerated. The complete proteinogram including the binding and non-binding dye proteins may be revealed by standard staining with bromophenol blue after removal of the supramolecular dye. An attempt for semi-automatic or automatic analysis of spots distributed in electrophoresis has been undertaken.