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  • Title: Determination of glucansucrase encoding gene in Leuconostoc mesenteroides.
    Author: Du R, Qiao X, Wang Y, Zhao B, Han Y, Zhou Z.
    Journal: Int J Biol Macromol; 2019 Sep 15; 137():761-766. PubMed ID: 31279888.
    Abstract:
    A glucansucrase encoding gene was cloned into pET-28a(+) vector and expression in Escherichia coli BL21(DE3). An about 160 kDa recombinant glucansucrase was purified with a yield of 50.73% and a 4.02-fold increase in activity. The 1464 amino acid residue enzyme belongs to the GH70 subfamily and shares 90% similarity with Leuconostoc sp. glucansucrase. The optimal temperature and pH were 30 °C and pH 5.5, and 80% of activity was retained after incubation at 10-30 °C and pH 5-7. Enzyme activity was strongly activated by Ca2+ and Mn2+ and inhibited by various metal ions and chemical agents, and a high affinity for sucrose (Km = 11.6 mM, Vmax = 8.1 mmol/(mL·min)). Circular dichroism (CD) and Raman spectra collectively indicated a high proportion of random coil structure.
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