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

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.


Keywords


lactic acid bacteria; Leuconostoc mesenteroides; dextransucrase; dextran

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DOI: http://dx.doi.org/10.2298/HEMIND160421046M

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