CC BY
26
UDC 541.49:546.791.6 +546.73
https://doi.org/10.15407/biotech13.04.060
BIODEGRADATION AND ANTIMICROBIAL ACTIVITY OF GUANIDINE-CONTAINING POLYETHYLENE
OXIDE HYDROGEL
G. O. Iutynska2 M. Ya. Vortman1 D. R. Abdulina2 Zh. P. Kopteva2 A. Ye. Kopteva2 A. V. Rudenko3 V. V. Tretyak3 V. N. Lemeshko1 V. V. Shevchenko1
institute of Macromolecular Chemistry of the National Academy of Sciences of Ukraine
2Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine
3Institute of Urology of the National Academy of Medical Sciences of Ukraine
E-mail: [email protected]
Received 11.05.2020 Revised 19.07.2020 Accepted 31.07.2020
The aim of the study was to determine the biodegradation of guanidine-containing polyethylene oxide hydrogel under the action of bacteria and to study its antimicrobial properties. The antimicrobial activity of newly synthesized polyethylene oxide hydrogel was studied by a disco-diffusion method. The enzymatic activities of bacteria were determined by spectrophotometry. To study the resistance of guanidine-containing polyethylene oxide hydrogel to microbial destruction, bacteria were grown in Tauson's liquid medium with the addition of meat peptone broth at a temperature of 28±2 °C. Changes in the chemical composition of the studied materials were analyzed by infrared Fourier spectroscopy and 1H NMR spectroscopy.
Guanidine-containing polyethylene oxide hydrogels were obtained by reacting oligooxyethylene glycol MW 6000 with toluene diisocyanate and guanidine-containing oligomer, which acted as a crosslinking, ion-containing and antibacterial-agent. The synthesized hydrogel showed antimicrobial activity against both gram-positive and gram-negative bacteria. The biodegradation of hydrogels under the action of various bacterial strains and enzymes that they synthesize was studied. The presence of the tested materials lead to a decrease in the enzymatic activity of bacteria in 2.5 times compared with the control.
Synthesized guanidine-containing polyethylene oxide hydrogel showed antimicrobial activity against the studied test cultures. The introduction of the studied materials into Tauson's environment as additional sources of carbon and energy helped to reduce the catalase and lipase activities of hydrocarbon-oxidizing bacteria. Under the influence of the studied bacteria, the hydrogels were destroyed up to 88.4% from the initial value.
Key words: Guanidine-containing polyethylene oxide hydrogel, antibacterial properties, gram-positive and gram-negative bacteria, catalase and lipase activities, biodegradation, IR spectra.
Biodegradation of various materials in the environment is important in terms of their protection because a lot of synthetic materials are stable for a long period. However, at the end of the life cycle, the materials must be involved in the natural cycle and decomposed. Expired materials and products are exposed to aggressive microorganisms. This confirms the
need to replace such materials on biodegradable ones [1-3]. Under natural conditions, the microbial attack on biodegradable material includes not one species, but the association of microorganisms both anaerobes and aerobes interact with each other and produce enzymes that are involved in the material destruction [4-6]. These microorganisms' activity is
influenced by environmental conditions, as well as the composition and chemical structure of the material. The bacteria of Cytophaga, Bacillus, Rhodococcus, Streptomyces, Mycobacterium, Pseudomonas genera and filamentous fungi of Aspergillus, Alternaria, Penicillium, Trichoderma genera, and others play the greatest role in the microbial destruction of materials [3].
In the current literature, there are a lack of data about enzymatic activity of polymer-destroying bacteria and mainly about on the activity of redox and hydrolytic enzymes in micromycetes that damage building materials, foams, rubbers, etc. [7, 8]. However, the lipolytic and catalase activities of bacteria, which result in microbial destruction of materials, reduce their strength and elasticity, remain poorly studied [7, 8].
In modern polymer chemistry, one of the promising areas is the obtaining and studying the properties of hydrogel systems, among which the polyethylene oxide hydrogels take a significant place. They are widely used in various fields of science and technology due to non-toxicity, high degree of swelling [911], as well as the ability to biodegradation [12-16]. One method of polyethylene oxide hydrogels producing is based on the reaction of urethane formation in the presence of a crosslinking agent. Particular importance is attached to the construction of pH-sensitive hydrogels, which are carried out by introducing a controlled number of ionic groups by copolymerization of acrylic derivatives of oligooxyethylene glycols with acrylic monomers containing such groups, or the introduction of ionic additives in the urethane formation reaction. The obtaining of hydrogels using derivatives of such a highly basic compound as guanidine, which has antimicrobial activity is of particular interest. Among guanidinium-containing polymers, polyhexamethylene guanidinium chloride attracts attention due to its biocide properties, which are important for practical use [17, 18]. It belongs to the fourth security class. The high reactivity of the guanidine fragment stimulated research in the direction of chemical modification of polyhexamethylene guanidine chloride in order to provide it with additional functionality. Its derivatives with methylene, acrylate groups are used for obtaining pH-sensitive hydrogels [19]. Guanidine-containing polyethylene oxide hydrogels have not yet been described in the literature. It can be assumed that such hydrogels should have
antibacterial and biodegradable properties as well as polyhexamethylene guanidine chloride will be low-toxic.
The aim of this work was to determine the antimicrobial properties of materials against bacteria isolated from pathogenic material of sick people and biodegradation of guanidine-containing polyethylene oxide hydrogel under the action of bacteria previously isolated and identified from damaged protective coatings of gas pipelines.
Materials and Methods
Materials. Diane epoxy oligomer DER-331 (DOW Chemical Company, Germany), mass fraction of epoxy groups 23.5% with a content of 0.6% hydroxyl groups was dehydrated by heating in vacuum for 2-6 hours at 80-90 °C and a final pressure of 266.6 pa. Guanidine hydrochloride (GD) (Sigma-Aldrich, USA, degree of purity 99.9%), medical ethanol rectification (96% ), oligooxyethylene glycol (OEG) MW 6000 (Sigma-Aldrich, USA, 99.9%) and toluene diisocyanate (TDI) — a mixture of isomers 2.4 and 2.6 (Sigma-Aldrich, USA, 99.9%) were used without further purification. Dimethylformamide (DMF) was purified by distillation.
Guanidine-containing oligomer (GO) was obtained at a temperature of 50 °C for two hours by reaction of 0.1 M of epoxy oligomer in 70% ethanol solution while stirring with an alcoholic solution of 0.2 M guanidine obtained immediately before the reaction. The yield of the final product was 95%.
Obtaining polyethylene oxide hydrogel was performed by melting oligooxyethylene glycol MW 6 000 and mixing it with guanidine-containing oligomer at 85 °C to homogenize the mixture at a molar ratio of the original components of OEG:GO-1.5:(1-1.5), then mixing with TDI at a molar ratio of the original components of 1.5:(1-1.5):2. The formation of the hydrogel was performed at 25 °C for one hour on a substrate, curing — at 80 °C for 4 hours. Two types of hydrogel are obtained, which differ in the molar content of guanidine-containing oligomer: hydrogel 1 — ratio of the initial components of OEG:GO:TDI (1.5:1:2) and hydrogel 2 — ratio of the initial components OEG:GO:TDI (1.5:1.5:2). The degree of crosslinking of the obtained hydrogel was determined by the content of the gel fraction in the Soxhlet apparatus in 93-95% acetone solution.
Disco-diffusion method was used to determine the bactericidal activity of the
original guanidine-containing oligomer and polyethylene oxide hydrogels against the bacteria Staphylococcus aureus 451, Escherichia coli 475, Pseudomonas mirabilis 494. These strains have been isolated from biomaterial of urogenital system of sick people and stored in the Laboratory of Bacteriological Research of the Institute of Urology of the National Academy of Medical Sciences of Ukraine. Also, hydrocarbon-oxidizing bacteria Pseudomonas pseudoalcaligenes 109, Rhodococcus erythropolis 102, Bacillus subtilis 138, which were previously isolated and identified by us from damaged protective coatings of gas pipelines [4, 6] and stored in the Collection of the Department of General and Soil Microbiology of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine were used. Guanidine-containing oligomer in the form of a 0.1 ml aqueous solution was used with standard paper disks with a diameter of 6 mm on the surface of meat-peptone agar inoculated with the appropriate bacterial test-culture. Polyethylene oxide hydrogels have been obtained based on guanidine-containing oligomer in the form of a film with a size of 10x10x0.5 mm. They were applied to the surface of meat-peptone agar inoculated with the appropriate test culture of bacteria. Incubation was performed for 18 hours at 28 ± 2 °C. Antimicrobial activity was calculated in diameter (mm) of the zones of growth inhibition of microorganisms. Antimicrobial activity was estimated by the following scale: zone 0-10 mm — non sensitive, 15-25 mm — sensitive, up to 25 mm — high sensitive.
Microbial degradation of guanidine-containing polyethylene oxidehydrogel was studied with the use of hydrocarbonoxidizing bacteria P. pseudoalcaligenes 109, R. erythro-polis 102 and B. subtilis 138 strains. Bacteria had grown in Tauson's liquid medium at 28 ± 2 °C [5] with the addition of 20% meat-peptone broth (MPB) as a source of Nitrogen and Carbon Test materials' samples with the sizes of 10x10x0.5 mm were weighed on electronic scales (ANG-200, AXIS) and sterilized with 72% solution of ethyl alcohol (30 min) and with UV rays with a wavelength of 256 nm (15 min) and were put in Tauson medium, inoculated with one of the above-mentioned bacterial strains. The amount of cells were 106 cells/ml calculated with serial dilution method. Variants with Tauson's medium. with materials without the addition of bacteria were used as control. The term of the experiment was 60 days. The destruction
of the samples was determined gravimetrically by loss of their mass. To perform this, after the exposure, the material samples were removed from the cultural liquid, air-dried, then weighed and the weight loss of control and test samples was determined. The amount of bacteria in the cultural liquid was determined with serial dilution method [20].
Enzymatic activity. To determine the effect of materials on the enzymatic activity of bacteria, cultural liquid was centrifuged for 20 min at 8 000 rp/min (2 000 g) with the use of Eppendorf 5810R centrifuge (Germany). Lipolytic activity was determined by reaction with p-nitrophenylpalmitate [21], catalase activity was determined using 0.03% hydrogen peroxide, which formed a stable colored complex with 4% molybdenum phosphate solution [22]. Enzymatic activity was determined in the supernatant spectrophotometrically on KFK-3 (Russian Federation). Total protein concentration in the cultural liquid was determined by the Lowry method. The specific activity of the studied enzymes was calculated according to the formulas specified in [18, 19] and was expressed in u-mg1 protein.
IR spectra of the investigated materials with Fourier transform were obtained on a spectrophotometer TENSOR 37 (BrukerOptik GmbH, Germany) in the spectral range of 6 000-400 cm1 in KBr tablets.
1H NMR spectra were recorded on a Varian VXR-400 MHz instrument in CDCI3 solution.
The degree of swelling of the obtained polyethylene oxide hydrogels in water for injections at pH 4.0, 7.0 and 9.8 at room temperature to constant values was measured by weighing on the torsion scales. The degree of swelling of the samples was calculated with the formula: a = (m - m0)/m0, where m0 is the mass of the sample before swelling, m is the mass of the swollen sample.
Statistical analysis of the obtained results was performed using the software package MS Excel 2010 and OriginPro 2016 (ver. B 9.3.226. www.originlab.com/). The obtained data were represented as M ± n (n = 3, at P < 0.05).
Results and Discussion
To obtain a polyethylene oxide hydrogel, a guanidine-containing oligomer of linear structure with terminal guanidine fragments has been synthesized by the reaction between a bifunctional aromatic oligoepoxide (MW 365) and guanidine at a molar ratio of 1:2 (Fig. 1).
OHl
Fig. 1. Scheme of synthesis of guanidine-containing oligomer
The structure of the synthesized guanidine-containing oligomer of linear structure was confirmed by IR and 1H NMR spectrometry by the following data:
IR (KBr) (vNH, OH), 3 156 cm-1 (8NH), 2 929 cm-1 (vCH), 2 963 cm-1 (vCH2), 2 869 cm-1(vCH3), 1 648 cm-1(vC=NandvNH), 1 4501 650 cm-1(vC=C C6H4), 1 100-1 300 cm-1(vC-O-C), the absorption bands of epoxy groups at 920 cm-1 were disappeared. In 1H NMR (CDCI3) spectra of guanidine-containing oligomer there are proton signals at 1.72 ppm (t, 3H, -CH3), 2.73 ppm -NH (NHCH2)), 2.58 ppm -CH2 (CH2CHOH), 3.58 ppm -OH (CH-OH)), 3.96 ppm -CH (CH-OH)), 6.8 ppm and 7.2 ppm -CH benzene ring, 8.4 ppm and 8.6 ppm -NH (-NH2 groups).
The synthesized guanidinium-containing oligoether with terminal guanidine moieties was used for the synthesis of polyethylene oxide hydrogels (1 and 2) as a crosslinking and antimicrobial agent. According to the results of microbiological studies, the obtained guanidine-containing oligoethers showed antimicrobial activity (Table 1; Fig. 2) against the studied bacterial test-cultures P. mirabilis 494, E. coli 475, P. pseudoalca-ligenes 109 and S. aureus 451.
The diameters of inhibition zone of bacterial growth were in the range of 13-20 mm. The largest zones of growth inhibition were observed for strains of R. erythropolis 102 and B. subtilis 138 (24-26 mm) under the influence of hydrogels 1 and 2. The obtained two variants of polyethylene oxide hydrogels showed antimicrobial activity against the studied test-cultures. The diameters of the zone growth inhibition for strains E. coli 475, P. mirabilis 494, S. aureus 451, P. pseudoalcaligenes 109 were in the range 1011 and 15-16 mm for hydrogel 1 and hydrogel 2, respectively. Hydrocarbon-oxidizing bacteria R. erythropolis 102 and B. subtilis138 were more sensitive to synthesized hydrogels because inhibition zones were 15-19 and 20-24 mm for hydrogel 1 and 2, respectively. Hydrogel 1 had low antibacterial activity
against the studied test cultures due to lower content of guanidine-containing oligoether in hydrogel 1, which acts as a crosslinking and antibacterial agent (Table 1). The scheme of synthesis of polyethylene oxide hydrogel is shown on Fig. 3.
The structure of the obtained hydrogels was confirmed by IR spectroscopy (Fig. 4).
IR spectra consists of (KBr) (vNH, OH) 3 156 cm-1, 2 949 (vCH), 2 896 cm-1 (vCH2), 2 868 cm-1 (vCH2), 1 648 cm-1 (SNH), (vC = N), (1 450-1 650) cm-1 (vC6H5), (1 1001 300)) cm-1 (vCOC).
The difference between the IR spectra of hydrogels 1 and 2 is the different ratio of the intensity of the absorption bands of the valence vibrations of the -CH bonds to the valence vibrations of the hydroxyl (-OH) and amino(-NH) groups. There was an increase in the absorption band of NH groups, because the molar content of GO in the hydrogel 2 was in 1.5 times higher than in hydrogel 1.
The degree of swelling of the polyethylene oxide hydrogels were obtained for various conditions, i.e. pH, temperature and swelling duration (Fig. 5).
The degree of swelling under different pH was in the range 8.05-11.5 (Fig. 5, a). The data show that both hydrogels are highly swellable and this parameter depends on the pH of the medium, the high degree of swelling 10.2-11.1 is observed under pH 4.0. The dependence of the degree of swelling of polyethylene oxide hydrogel 2 in water on temperature (Fig. 5, b) showed that high values 14 is observed at 56 °C.The study of the duration of swelling of hydrogel 2 (Fig. 5, c) showed that under pH 4.0 and pH 7.0 during 180-210 min there was complete swelling and values were 11.1 and 9.0, respectively.
The hydrocarbon-oxidizing bacteria P. pseudoalcaligenes 109, R. erythropolis 102 and B. subtilis 138 produced extracellular poly-and lipopolysaccharides, due to which these bacteria adhere to the surface of protective materials, ensuring their rapid reproduction and promoting survival of bacterial population
Table 1. Antimicrobial properties of the original oligomer and obtained polyethylene oxide hydrogels
Bacterial strains Growth inhibition zone diameter, mm
Guanidine-containing oligoether Guanidine-containing hydrogel 1 Guanidine-containin ghydrogel 2
Gram-negative bacteria
Escherichia coli 475 14 i 0.20 10 i 0.08 16 i 0.20
Pseudomonas mirabilis 494 13 i 0.15 11 i 0.13 16 i 0.15
P. pseudoalcaligenes 109 15 i 0.17 10 i 0.10 15 i 0.17
Gram-positive bacteria
Rhodococcus erythropolis 102 24 i 0.32 15 i 0.15 20 i 0.32
Bacillus subtilis 138 26 i 0.25 19 i 0.20 24 i 0.25
Staphylococcus aureus 451 20 i 0.08 11 i 0.17 15 i 0.08
Notes: 0-10 mm non sensitive microorganism; 15-25 mm — sensitive, up to 25 mm — high sensitive (n = 3)
Fig. 2. Growth inhibition zone diameter of the Rhodococcus erythropolis 102 (a) and Bacillus subtilis 138 (b)
under the influence of polyethylene oxide hydrogels
1 — control (distilled water); 2 — hydrogel 1; 3 — hydrogel 2; 4 — initial guanidine-containing oligomer
Fig 3. Scheme of polyethylene oxide hydrogel synthesis
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in extreme conditions. The identified exopolysaccharides differed in monosaccharide composition and fatty acid content. Analysis of the fatty acid composition of the exopolymer complex of bacteria revealed the presence of saturated, unsaturated fatty acids and hydroxyl- acids with a carbon chain length of 11 to 18 carbon atoms [7].
In Tauson's liquid medium (with the addition of MPB, inoculated with bacterial strains and without hydrogels (control), increasing the number of hydrocarbon-oxidizing bacteria to 109 cells/ml over 60 days of the experiment was observed. In the experimental variants in the presence of the samples of studied hydrogels it was observed
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n = 3, data marked with * are statistically significant (P < 0.05)
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the reduced number of bacteria in 2-3 times compared to the initial titer.
Thus, we studied the influence of obtained hydrogels on the enzymatic activity of bacteria. Bacterial cultures in the presence of the studied hydrogels had different enzymatic activity depending on the hydrogels (Fig. 6).
The specific catalase activity in the control (nutrient media with bacteria and without the hydrogels) was higher than in presence of bacteria and materials. The highest specific catalase activity was in B. subtilis 138 and R. erythropolis 102 (5.3 ± 0.5 and 5.2 ± 0.4 u-mg 1 protein, respectively), but in the presence of the studied hydrogels enzymatic activity was decreased in 1.1-1.9 times for different bacterial cultures. Catalase activity of P. pseudoalcaligenes 109 in the presence of hydrogels 1 and 2 was significantly lower from 2.4 to 5.0 times, respectively (Fig. 6, a). In Tauson's medium without the introduction of materials, higher lipase as well as catalase activities were found in B. subtilis 138 (19.25 ± 2.1 u-mg-1 protein) and R. erythropolis 102 (18.6 ± 1.9 u-mg"1 protein), compared with other bacterial cultures. In the presence of the studied hydrogels 1 and 2 lipase activities decreased in 1.4-1.8 times, respectively. The specific lipase activity of P. pseudoalcaligenes 109 with guanidine-containing hydrogels in cultural liquid was decreased in 1.3-1.9 times compared to control. In the presence of hydrogel 1, compared to hydrogel 2 the lipase activity of bacterial test-cultures was lower (7.8 ± 1.4-12.7 ± 2.3 u-mg-1 protein).
It is known from literature data that changes in catalase activity are one of the
indicators of processes intensity of organic matter oxidation [23, 24]. There is a close correlation between the reduction degree of catalase activity of destructive strains and the efficiency of oil consumption. Active destructive bacteria have characterized in increasing their amount and decreasing catalase activity in contact with the petroleum products [25]. According to several reports, hydrocarbon-oxidizing bacteria activate lipase activity of soils, while in parallel with the activation of lipolysis, there is an increase in the number of hydrocarbon-oxidizing bacteria and a decrease of petroleum products [25].
The utilization of different materials after their use is an important aspect of the problem of materials biodamages. Significant indicator of material degradation is loss of the sample mass due to bacterial action (Table 2).
After 60 days of the experiment, the largest weight loss 0.35 ± 0.1 g (85.4% from the initial mass) of hydrogel 1 samples was observed under the influence of B. subtilis 138. In the variant of the experiment with hydrogel 2, the weight loss of the samples under the influence of bacteria was from 0.12 ± 0.01 to 0.22 ± 0.05 g (59.5-88.4%). In the control, the weight loss of hydrogel 1 was 0.06 ± 0.01 g (17%).
Degradation of guanidine-containing hydrogel materials under the influence of heterotrophic bacteria was confirmed by IR spectroscopy. As in the control hydrogels, and in the IR spectra of the hydrogels under the influence of bacterial test-cultures there
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Fig. 6. Specific catalase (a) and lipase (b) activity of hydrocarbon-oxidizing bacteria in the presence
of guanidine-containing hydrogels
C — control (without the introduction of materials), 1 — hydrogel 1; 2 — hydrogel 2; 109 — P. pseudoalcaligenes 109; 102 — R. erythropolis 102; 138 — B. subtilis 138; n = 3; data marked with * are
statistically significant (P < 0.05).
b
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Table 2. Changes in the mass of samples of guanidine-containing polyethylene oxide hydrogels in the presence of hydrocarbon-oxidizing bacteria
Experiment variant, g Loss of sample mass
% from the initial mass
Hydrogel 1 P. pseudoalcaligenes 109 0.26 ± 0.09 75.0
R. erythropolis 102 0.19 ± 0.04 63.1
B. subtilis 138 0.35 ± 0.10 85.4
Control without bacteria and with material 0.06 ± 0.01 17.0
Hydrogel 2 P. pseudoalcaligenes 109 0.12 ± 0.01 59.5
R. erythropolis 102 0.22 ± 0.05 88.4
B. subtilis 138 0.24 ± 0.06 80.0
Control without bacteria and with material 0.03 ± 0.01 38.5
Notes: n = 3, P < 0.05.
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50010001 JOG 2000 2500 3000 35004000
Wavelength, cm1
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50010001500 2000 2500 3000 35004000
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Fig. 7. IR spectra of polyethylene oxide hydrogels samples under the influence of hydrocarbon-oxidizing bacteria: a — hydrogel 1; b — hydrogel 2; I — initial hydrogel 1; II — initial hydrogel 2; 1 — control; 2 — B. subtilis 138; 3 — R. erythropolis 102
а
appeared next absorption bands (vNH, OH), 3 156 cm-1 (SNH), 2 949 cm-1(vCH), 2 896 cm-1 (vCH2), 2 868 cm-1 (vCH2), 1 648 cm-1 (vC = N), (1 450-1 650) cm-1 (vC6H5), 1 100-1 300 cm-1 (vCOC) (Fig. 7).
The difference was that for hydrogels 1 and 2 under the influence of R. erythropolis 102 and B. subtilis 138 after 60 days, decreased the absorption band of 3500 cm-1 decreased, which was responsible for the valence fluctuations of hydroxyl (-OH) and amino (-NH) groups (Fig. 5). Because crosslinking of polyethylene oxide hydrogels occurs through the formation of amide bonds that are resistant to hydrolysis and biodegradation, it can be assumed that the presence of a significant number of secondary amino groups in the crosslinking agent contributes as a catalyst to biodegradation of hydrogels, especially under influence of the R. erythropolis 102.
The mechanism of biocidal action of po-lyguanidines is similar to quaternary ammonium compounds and is of membrane-toxic nature: guanidine polycations are adsorbed on the negatively charged surface of bacterial cells; diffuse through the cell wall; bind to acidic phospholipids, proteins of the cytoplasm membrane, which causes its breakage. As a result, the microbial cell dies. The increase in the biocidal activity of PHMGC as compared to the low-molecular biocides is due to the cooperative interaction of the adjacent guanidine polycation group links with the microbial cell. The increase in the activity of polyguanidines as compared to quaternary ammonium salts (QAS) is also due to the peculiarities of the guanidine group structure: unlike QAS cation, in which a large positive charge is localized on a single nitrogen atom, guanidine cation has a positive charge distributed between three nitrogen atoms, and additionally delocalized according
to the ст-links system. It is well known that guanidinium polymers are less toxic than guanidine and belong to the third class of hazard [17, 18].
The antimicrobial properties and biodegradation of newly synthesized guanidine-containing polyethylene oxide hydrogels, in which the linear guanidine-containing oligomer simultaneously acts as a crosslinking, ion-containing and antimicrobial agent, were studied. The synthesized hydrogel showed antimicrobial activity against the studied test cultures of bacteria. The largest zones of growth inhibition were observed for S. aureus 451, R. erythropolis 102, B. subtilis 138. The introduction of the studied materials into the Tauson's medium as additional sources of Carbon and energy, contributed to the reduction of catalase and lipase activities of hydrocarbon-oxidizing bacteria. Under the influence of the studied bacteria, destruction of hydrogels, which is appeared in a decreasing of the studied samples mass, was observed. Infrared spectroscopy showed that the degradation of guanidine-containing hydrogels is affected by the presence of a significant number of secondary amino groups in the crosslinking agent, which can serve as catalysts for the biodegradation of hydrogels.
The research was carried out within the framework of the projects of the National Academy of Sciences of Ukraine "Functional activity and prospects of use in biotechnology soil microorganisms from natural, man-caused and agro systems" (2016-2019, State registration number GDR 0116U006319) and "Development of methods of synthesis of monomers containing perfluorated and highly basic nitrogenic and other functional groups and fragments" (2018-2022, GDR0118U002055).
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Б1ОДЕГРАДАЦ1Я ТА АНТИМ1КРОБНА
АКТИВН1СТЬ ГУАН1ДИНВМ1СНОГО ПОЛ1ЕТИЛЕНОКСИДНОГО Г1ДРОГЕЛЮ
Г. 0.1утинська2, М. Я. Вортман1, Д. Р. АбдулЬна2, Ж. П. Коптева2, Г. G. Коптева2, А. В. Руденко3, В. В. Третяк3, В. М. Лемешко1, В. В. Шевченко1
Институт xiMii високомолекулярних сполук НАН Укра1ни 21нститут мшробмлогй i вiрусологil iM. Д.К. Заболотного НАН Укра1ни, Ки1в 31нститут урологи НАМН Укра1ни, Ки1в
E-mail: [email protected]
Метою роботи було визначити бшрозкла-дання гуашдинвм^ного полiетиленоксидного гiдрогелю пiд дieю бактерш та вивчити його антимiкробнi властивость
Антимiкробну активнiсть новосинтезова-них полiетиленоксидних гiдрогелiв вивчали диско-дифузiйним методом. Ензиматичну актившсть бактерiй визначали спектрофо-тометрично. Стшшсть гуанiдинвмiсного по-лiетиленоксидного идрогелю до м^робно! деструкцп перевiряли в рщкому середовишД Таусона з тест-культурами бактерш з дода-ванням м'ясо-пептонного бульйону за тем-ператури 28±2 °С. Змiни хiмiчного складу дослщжених матерiалiв вивчали методом шфрачервоно1 та 1Н ЯМР-спектроскопп.
Гуанiдинвмiсний полiетиленоксидний гидрогель було отримано шляхом взаемодп оль гооксiетиленглiколю ММ 6000 з толу!лендь зоцианатом i гуанiдинвмiсним ол^омером, який вiдiграe роль зшиваючого, ^нвм^ного та протимiкробного агента. Синтезований пдрогель виявляв антимiкробну активнiсть щодо грампозитивних та грамнегативних бактерiй.
Внесення в середовище Таусона досль джуваних матерiалiв як додаткових джерел вуглецю та енерги призводило до знижен-ня каталазно1 та лшазно1 активност вугле-водньоокиснювальних бактерiй. За впливу дослщжуваних бактерiй вiдбувалась деструк-щя гiдрогелiв до 88,4% вiд початкового зна-чення.
Ключовi слова: гуашдинвм^ний полiетиле-ноксидний гiдрогель, антибактерiальнi власти-воси, грампозитивнi та грамнегативнi бактери, каталазна та лiпазна активнiсть, бiодеградацiя, 1Ч-спектри.
БИОДЕГРАДАЦИЯ И АНТИМИКРОБНАЯ АКТИВНОСТЬ ГУАНИДИНСОДЕРЖАЩЕГО ПОЛИЭТИЛЕНОКСИДНОГО ГИДРОГЕЛЯ
Г. А. Иутинская2, М. Я. Вортман1, Д. Р. Абдулина2, Ж. П. Коптева2, Г. Е. Коптева2, А. В. Руденко3, В. В. Третяк3, В. Н. Лемешко1, В. В. Шевченко1
1Институт химии высокомолекулярных соединений НАН Украины, Киев, Украина 2Институт микробиологии и вирусологииим.
Д. К. Заболотного НАН Украины, Киев 3Институт урологии НАМН Украины, Киев
E-mail: [email protected]
Целью данной работы было определение биоразложения гуанидинсодержащего полиэтиле-ноксидного гидрогеля под действием бактерий и изучение его антимикробных свойств.
Антимикробную активность новосинтезиро-ванного полиэтиленоксидного гидрогеля изучали диско-диффузионным методом. Энзиматическую активность бактерий определяли спектрофотоме-трически. Устойчивость гуанидинсодержащего полиэтиленоксидного гидрогеля к микробной деструкции проверяли в жидкой среде Таусона с тест-культурами бактерий с добавлением мясо-пептонного бульона при температуре 28 ± 2 °С. Изменения химического состава исследованных материалов изучали методом инфракрасной Фурье- и 1Н ЯМР-спектроскопии.
Гуанидинсодержащий полиэтиленоксид-ный гидрогель был получен путем взаимодействия олигооксиэтиленгликоля ММ 6000 с толуилендиизоцианатом и гуанидинсодер-жащим олигомером, который выступает как сшивающий, ионсодержащий и противоми-кробный агент. Синтезированный гидрогель проявлял антимикробную активность в отношении грамположительных и грамотрица-тельных бактерий.
Внесение в среду Таусона исследуемых материалов как дополнительных источников углерода и энергии снижало каталазную и липазную активность углеводородоокисляющих бактерий. Под влиянием исследуемых бактерий происходила деструкция гидрогелей до 88,4% от первоначального значения.
Ключевые слова: гуанидинсодержащий поли-этиленоксидный гидрогель, антибактериальные свойства, грамположительные и грамо-трицательные бактерии, каталазная и ли паз ные активности, биодеградация, ИК-спектры.
