Mikrobiol. Z. 2017; 79(5):13-22. Ukrainian.
doi: https://doi.org/10.15407/microbiolj79.05.013

Influence of Divalent Cations on Synthesis of Nocardia vaccinii ІMV B-7405
Surfactants With High Antimicrobial and Anti-Adhesion Activity

Pirog T.P.1,2, Nikituk L.V.1, Shevchuk T.A.2

1National University of Food Technologies
68 Volodymyrska Str., Kyiv, 01601, Ukraine

2Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

Aim. To study the effect of divalent cations on NADP+-dependent glutamate dehydrogenase activity (key enzyme of biosynthesis of Nocardia vaccinii IMV B-7405 surface-active aminolipids) followed by appropriate modification of medium composition for synthesis of surfactants with high anti-adhesive and antimicrobial activity. Methods. The NADP+-dependent glutamate dehydrogenase activity (EC 1.4.1.4) in the cell-free extract was analyzed for the formation of glutamate in the oxidation of NADPH at 340 nm. Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2 : 1). Antimicrobial against bacteria and yeast properties of the surfactant was determined by index of the minimum inhibitory concentration (MIC). The number of attached cells was determined spectrophotometrically. Results. It was established that in the presence of 5 − 10 mM calcium cations NADP+-dependent glutamate dehydrogenase activity in the cell-free extract of N. vaccinii IMV B-7405 increased to 1.3 − 1.7 times in comparison with that without Ca2+. Increase from 0.1 to 0.4 g/l CaCl2 × 2H2O concentration in cultivation medium of IMV B-7405 strain was accompanied by rise of NADP+-dependent glutamate dehydrogenase activity in 1.5 times, as well as increasing antimicrobial and antiadhesive activity of synthesized surfactants. Minimum inhibitory concentration against some bacteria and yeasts surfactants synthesized in a medium with a heightened content of Ca2+ was in 1.8 − 13 times and adhesion test cultures on abiotic surfaces treated with such surfactants − an average of 7 − 39 % lower as compared to indicators for the surfactant produced in the base medium. Conclusions. The obtained results indicate the possibility of regulation of the antimicrobial and anti-adhesive activity of microbial surfactants with the change in cultivation medium of cations content – potential activators and/or inhibitors of key enzymes biosynthesis of surfactant components responsible for these biological properties.

Key words: Nocardia vaccinii ІMV B-7405, surfactants, antimicrobial and antiadhesive activity, activity of key enzymes biosynthesis.

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