Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate

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Miona G. Miljković
Slađana Z. Davidović
Slavko Kralj
Slavica S. Šiler-Marinković
Mirjana D. Rajilić-Stojanović
Suzana I. Dimitrijević-Branković

Abstract

The production of dextransucrase (DS) by Leuconostoc mesenteroides T3, novel isolate from water kefir grain, was studied and optimized. Bacterial supernatant reached activity of 3.1 U/ml when the culture was grown at 23 °C and under static culture condition using classical Tsuchiya medium for DS production. The increase of sucrose concentration to 7% led to an increase of DS activity by 52% compared to the control. Medium with 2% beef extract and 1% yeast extract resulted in 4.52 U/ml, which was 47% higher than in the control (with 2% yeast extract). Finally, the increase of K2HPO4 concentration from 2 to 3% resulted in the increased enzyme activity by 28%. Enzyme purified by polyethylene glycol 400 fractionation displayed maximum activity at 30 °C and pH 5.4. Zymogram analysis confirmed the presence of DS of approximately 180 kDa. The addition of divalent cations Ca2+, Mg2+, Fe2+ and Co2+ led to a minor increase of DS activity, while the addition of Mn2+ was the most prominent with 73% increase. These findings classify dextransucrase from Leuconostoc mesenteroides T3 as promising candidate for production of dextran, which has numerous applications in various industries.

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Miljković, M. G., Davidović, S. Z., Kralj, S., Šiler-Marinković, S. S., Rajilić-Stojanović, M. D., & Dimitrijević-Branković, S. I. (2017). Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate. HEMIJSKA INDUSTRIJA (Chemical Industry), 71(4), 351–360. https://doi.org/10.2298/HEMIND160421046M
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