Особенности взаимодействия лизоцима с блок-сополимерами PGLU–PEG

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Abstract

Исследовано взаимодействие лизоцима белка куриного яйца с блок-сополимерами поли(L-глутаминовой кислоты натриевой соли) и полиэтиленгликоля (PGLU10–PEG, PGLU100–PEG) в водной среде и на границе вода–воздух. При проведении исследований применяли широкий спектр физико-химических методов: турбидиметрия, тензиометрия, флуориметрия, КД-спектроскопия, электрофоретическое светорассеяние, просвечивающая электронная микроскопия. Обнаружено, что на границе вода–воздух возможно образование смешанных адсорбционных слоев при мольных соотношениях блок-сополимер: фермент, не превышающих 2 : 1. В водной среде возможно образование комплексов блок-сополимера PGLU10–PEG и лизоцима со структурой типа ядро–оболочка и комплексов блок-сополимера PGLU100–PEG: лизоцим состава 1 : 1 или 2 : 1 (по молям). Возможность регулировать свойства продуктов взаимодействия фермента и блок-сополимеров позволяет разрабатывать стратегии получения антибактериальных препаратов.

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About the authors

Л. Ю. Филатова

Московский государственный университет имени М. В. Ломоносова

Author for correspondence.
Email: luboff.filatova@gmail.com

Химический факультет

Russian Federation, Ленинские горы, д. 1, стр. 3, Москва, 119991

Н. Г. Балабушевич

Московский государственный университет имени М. В. Ломоносова

Email: luboff.filatova@gmail.com

Химический факультет

Russian Federation, Ленинские горы, д. 1, стр. 3, Москва, 119991

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Supplementary files

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2. Fig. 1. Isotherms of surface tension for solutions of block copolymers PGLU10–PEG (a), PGLU100–PEG (b) in a free state and mixed with lysozyme (7.0 · 10-6 M) in an aqueous medium at 25 ° C. Gray icons are lysozyme, black icons are block copolymers, white icons are mixtures of lysozyme and block copolymers.

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3. Fig. 2. Dependence of the pH of the medium of mixtures of lysozyme with block copolymers on the concentration of the block copolymer. The black columns are lysozyme with PGLU10–PEG, the white columns are lysozyme with PGLU100–PEG.

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4. Fig. 4. TEM images and particle size distributions for PGLU100–PEG block copolymer (a) and lysozyme with PGLU100- PEG block copolymer, Z 18:1 (b).

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5. Fig. 5. TEM images and particle size distributions for PGLU10-PEG block copolymer (a) and lysozyme with PGLU10-PEG block copolymer, Z 18:1 (b).

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6. Fig. 6. The effect of block copolymers on the position of the maximum intensity of the fluorescence of lysozyme (a), vli‑ the effect of block copolymers on the intensity of the maximum fluorescence of lysozyme (b). Black icons are lysozyme with PGLU10–PEG, white icons are lysozyme with PGLU100–PEG. λmax is the wavelength of the maximum fluorescence intensity; RFU0 is the number of units of lysozyme fluorescence at a wavelength of 339 nm (at λmax); RFU is the number of units of intensity of fluorescence of lysozyme with a block copolymer at a wavelength of 339 nm. RFU0/RFU (λmax) is the ratio of the intensity of the maximum fluorescence of free lysozyme to the intensity of fluorescence of lysozyme with a block copolymer at a wavelength of 339 nm.

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7. Fig. 7. CD spectra of free lysozyme and enzyme in the composition of associates with block copolymers PGLU10–PEG, PGLU100–PEG. The black line is a free lysozyme, the black dotted line is a lysozyme with PGLU100–PEG (Z 18 : 1), the gray dotted line is a lysozyme with PGLU10–PEG (Z 18 : 1).

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8. Figure 8. The effect of block copolymers on lysozyme activity. The black columns are lysozyme with PGLU10– PEG, the white columns are lysozyme with PGLU100–PEG.

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