The gene encoding an exo-inulinase was cloned into pET-28a(+) from an inulin-degrading Paenibacillus sp. Lfos16 and expressed in Escherichia coli BL21(DE3). The recombinant exo-inulinase was purified using nickel column affinity chromatography and verified by SDS-PAGE. The molecular weight of recombinant exo-inulinase was 87 kDa with the specific activity of 348.30 U/mg protein. Its activities on inulin and sucrose were 259.37 U/mL and 592.16 U/mL, respectively, with an I/S value of 0.438, and fructose was the main product from inulin. The recombinant exo-inulinase had the optimal reaction temperature and pH of 40 ℃ and 6.0, respectively. It was stable when the temperature was lower than 30 ℃. It was significantly inhibited by Ag+, Cu2+, Mn2+, Zn2+, Hg2+ and Fe3+. The Km and Vmax of recombinant exo-inulinase on inulin were 19.28 mg/mL and 0.18 mg/(min·mL), respectively.
MA Jun
,
AN Qikun
,
TANG Wenzhu
. Heterelogous expression, purification and characterization of an exo-inulinase from Paenibacillus sp. Lfos 16[J]. Food and Fermentation Industries, 2019
, 45(4)
: 25
-30
.
DOI: 10.13995/j.cnki.11-1802/ts.018537
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