Abstract
To construct a high-performance engineered endo-inulinase for fructo-oligosaccharides (FOS) production from inulin, an inulin binding module (IBM) was fused into either N- or C-terminal of an endo-inulinase. After heterologous expression, purification and characterization, the C-terminal fusion one (Eninu-IBM) with better activity, thermostability and inulin binding ability was employed for high-temperature in situ inulin hydrolysis in a 10-L fermentor. During this process, Eninu-IBM was first efficiently produced by the yeast cells at 28 °C for 96 h, and subsequently 1600 g unsterilized inulin per liter fermentation liquor was directly supplemented into the bioreactor for FOS production at 60 °C for 2 h. Finally, high purity of FOS (91.4%) were obtained with FOS titer, yield and productivity of 717.3 g/L, 0.912 gFOS/gInulin and 358.6 g/L/h, respectively. The in vitro prebiotic assay indicated that the final FOS products with main polymerization degrees of 3–5 were preferably fermented by beneficial bifidobacteria and lactobacilli.
Published Version
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