期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 91, 期 -, 页码 123-131出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ijbiomac.2016.05.019
关键词
L citreum SK24.002 glucansucrase; Characterization; De novo synthesis
资金
- National Natural Science Foundation of China [31000764, 31230057]
- International Cooperative Program of Jiangsu Province [BZ2012031]
- Science & Technology Pillar Program of Jiangsu Province [BE2012613, BE2013647, BE2014703]
- Fundamental Research Funds for the Central Universities [JUSRP51616B]
The cell-associated glucansucrase from Leuconostoc citreum SK24.002 was isolated, purified, characterized and used for de novo synthesis of alpha-glucan and acceptor-products. The final specific glucansucrase activity was 1.4 U/mg protein with 13.2-fold purification. The obtained glucansucrase had a molecular weight of 186 kDa, T-m of 61.7 degrees C and Delta H of 176.7 kJ/mol. The enzyme showed maximum activity at pH 5.0-6.0 and 45 degrees C. The enzyme activity was enhanced by Ca2+, Mn2+ or Co2+ ions, whereas the activity decreased as the methanol, ethanol, n-butanol, DMSO or isopropanol concentration increased. The chemical inhibitors including BD, DTNB, EDC or NBS also significantly inhibited enzyme activity. K-m, V-max and k(cal) of glucansucrase were 10.9 mM, 3.6 U/mg and 306.61/s, respectively. For de novo synthesis from sucrose, alpha-glucan polymer with molecular weight of 1.5 x 10(7) g/mol and maltose acceptor products (trisaccharide, tetrasaccharide and pentasaccharide) were obtained by glucansucrase via glucosyltransfer reactions, respectively. (C) 2016 Elsevier B.V. All rights reserved.
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