vestibИзвестия Национальной академии наук Беларуси. Серия биологических наукProceedings of the National Academy of Sciences of Belarus, Biological Series1029-89402524-230XThe Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1029-8940-2018-63-3-307-315vestib-379Research ArticleСтатьиВЛИЯНИЕ УСЛОВИЙ КУЛЬТИВИРОВАНИЯ НА АНТИМИКРОБНУЮ И АНТИАДГЕЗИВНУЮ АКТИВНОСТЬ ПОВЕРХНОСТНО-АКТИВНЫХ ВЕЩЕСТВ БАКТЕРИЙ РОДОВ ACINETOBACTER, RHODOCOCCUS И NOCARDIAINFLUENCE OF CULTIVATION CONDITIONS ON ANTIMICROBIAL AND ANTI-ADHESIVE ACTIVITY OF SURFACTANTS OF BACTERIA OF ACINETOBACTER, RHODOCOCCUS AND NOCARDIA GENERAПирогТ. П.PirogT. P.

д-р биол. наук, вед. науч. сотрудник, профессор, заведующий кафедрой

D. Sc. (Biol.), Leading researcher, Professor, Head of the Department

tapirog@nuft.edu.uaШевчукТ. А.ShevchukT. A.

вед. инженер

Leading Engineer

НикитюкЛ. В.NikitukL. V.

аспирант

Postgraduate student

liya.nikityuk@ukr.netЛуцайД. А.LutsaiD. A.

студент

Student

lutayda0@ukr.netПалийчукО. И.PaliichukO. I.

аспирант

Postgraduate student

olesiapaliichuk@gmail.comНациональный университет пищевых технологий, КиевNational University of Food Technologies, KyivИнститут микробиологии и вирусологии им. Д. К. Заболотного НАН Украины, КиевZabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Kyiv201825082018633307315Copyright © Пирог Т.П., Шевчук Т.А., Никитюк Л.В., Луцай Д.А., Палийчук О.И., 20182018Пирог Т.П., Шевчук Т.А., Никитюк Л.В., Луцай Д.А., Палийчук О.И.Pirog T.P., Shevchuk T.A., Nikituk L.V., Lutsai D.A., Paliichuk O.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestibio.belnauka.by/jour/article/view/379

Микробные поверхностно-активные вещества (ПАВ) являются продуктами мультифункционального назначения, поскольку они способны не только снижать поверхностное натяжение на границе раздела фаз и эмульгировать различные субстраты, но и проявлять антимикробную и антиадгезивную активность (в том числе и способность к разрушению биопленок). Однако в различных условиях культивирования продуцентов состав ПАВ и их свойства могут изменяться. Одним из подходов к повышению антимикробной и антиадгезивной активности ПАВ может быть увеличение в среде культивирования продуцентов содержания активаторов ключевых ферментов биосинтеза аминолипидов – наиболее эффективных антимикробных агентов. Активаторами НАДФ+-зависимой глутаматдегидрогеназы у Acinetobacter calcoaceticus IМВ В-7241 являются катионы кальция, магния и цинка, у Rhodococcus erythropolis ІМВ Ас-5017 и Nocardia vaccinii IМВ В-7405 − катионы кальция. Цель работы – исследовать антимикробную и антиадгезивную активность ПАВ, синтезированных A. calcoaceticus IМВ В-7241, R. erythropolis IМВ Ас-5017 и N. vaccinii ІМВ В-7405 в среде с повышенным содержанием активаторов НАДФ+-зависимой глутаматдегидрогеназы. ПАВ экстрагировали из супернатанта культуральной жидкости смесью хлороформа и метанола (2:1). Антимикробную активность ПАВ определяли по показателю минимальной ингибирующей концентрации (МИК), антиадгезивную – спектрофотометрическим методом. О степени разрушения биопленки судили по разнице между количеством адгезированных клеток в необработанных и обработанных ПАВ лунках полистиролового планшета с предварительно сформированной биопленкой и выражали в процентах. Установлено, что дополнительное внесение СaCl2 (0,1 г/л) в среду культивирования R. erythropolis IМВ Ac-5017, повышение концентрации этой соли до 0,4 г/л в среде для выращивания N. vaccinii ІМВ В-7405, а также добавление СaCl2 (0,1 г/л), увеличение содержания MgSO4 ·7H2O до 0,2 г/л или внесение Zn2+ (38 мкМ) в среду культивирования A. calcoaceticus IМВ В-7241 сопровождалось синтезом ПАВ, МИК которых по отношению к тест-культурам были в 1,2−13 раз ниже, их адгезия на абиотических поверхностях, обработанных такими ПАВ, – в среднем на 10–40 % ниже, а степень разрушения биопленок – на 7–20 % выше по сравнению с показателями, установленными для ПАВ, полученных на базовой среде. Приведенные данные свидетельствуют о возможности регуляции антимикробной и антиадгезивной активности микробных ПАВ в процессе культивирования продуцента.

Microbial surfactants are multifunctional products because they cannot only reduce the surface tension at the interface and emulsify various substrates, but also display antimicrobial and anti-adhesion activity (including the ability to destroy biofilms). However, under various conditions of producer’s cultivation the surfactant composition and their properties can vary. One of the approaches to increasing antimicrobial and anti-adhesion activity of the surfactant can be an increase in medium of producer cultivation content of activators of key enzymes biosynthesis of aminolipids − the most effective antimicrobial agents. Activators of NADP+-dependent glutamate dehydrogenase in Acinetobacter calcoaceticus IMV B-7241 are cations of calcium, magnesium and zinc, Rhodococcus erythropolis IMV Ac-5017 and Nocardia vaccinii IMV B-7405 – calcium.

Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2:1). Antimicrobial activity of surfactants was determined by index of minimum inhibitory concentration (MIC), antiadhesive − by spectrophotometry. The degree of biofilm destruction was determined as difference between the number of adhered cells in untreated and treated with surfactant holes of polystyrene immunological plate containing pre-formed biofilm of test cultures and was expressed as a percentage. It was found that addition of CaCl2 (0.1 g/l) into medium cultivation of R. erythropolis IMV Ac-5017, increasing concentration of this salt to 0.4 g/l in medium for N. vaccinii IMV B-7405 growth, introduction of CaCl2 (0.1 g l) and increasing MgSO4 ·7H2O content to 0.2 g/l, or adding Zn2+ (38 μM) into medium cultivation of A. calcoaceticus IMV B-7241 was accompanied by synthesis of surfactants MICs of which against test cultures were 1.2–13 times lower, their adhesion on abiotic surfaces treated with such surfactants was on average 10−40 % lower, and the degree of biofilms destruction was 7−20 % higher than indicators established for surfactants obtained on the base medium. The obtained data indicate the possibility of regulating antimicrobial and anti-adhesion activity of microbial surfactants under producer cultivation.

 

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The authors declare that there are no conflicts of interest present.