Purification and Characterization of Dextransucrase from Leuconostoc Mesenteroides Nrrl B-1149
نویسندگان
چکیده
Leuconostoc mesenteroides NRRL B-1149 produces extracellular dextransucrase which in this study was purified using different concentrations of polyethylene glycol (PEG). The dextran produced by this enzyme is unique in that it contains α(16) and α-(13) linkages which have clinical applications. The cell free supernatant with 0.9 U/mg enzyme specific activity was subjected to fractionation by PEG-400 and PEG-1500. The 33% PEG-400 gave dextransucrase with specific activity of 9.2 U/mg and 10 fold purification and the 15% PEG-1500 gave dextransucrase with maximum specific activity of 15 U/mg and 17 fold purification in a single step. The purified enzyme showed multiple molecular forms on denaturing SDSPAGE with three prominent bands. The purified dextransucrase confirmed the presence of glucan, after in-situ activity detection by Periodic acid Schiff’s staining after running under denaturing SDS-PAGE. The three bands that appeared on denaturing SDS-PAGE stained with silver nitrate solution, corresponded to the three activity bands.
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