Abstract
Fructooligosaccharides are well-known carbohydrate molecules that exhibit good probiotic activity and are widely used as sweeteners. Inulin-type fructooligosaccharides (IFOs) can be synthesized from sucrose using inulosucrase. In this study, cross-linked enzyme aggregates (CLEAs) of Lactobacillus reuteri 121 inulosucrase (R483A-LrInu) were prepared and used as a biocatalyst for IFOs production. Under optimum conditions, R483A-LrInu CLEAs retained 42% of original inulosucrase activity. Biochemical characterization demonstrated that the optimum pH of inulosucrase changed from 5 to 4 after immobilization, while the optimum temperature was unchanged. Furthermore, the pH stability and thermostability of the R483A-LrInu CLEAs was significantly improved. IFOs product characterization indicated that the product specificity of the enzyme was impacted by CLEA generation, producing a narrower range of IFOs than the soluble enzyme. In addition, the R483A-LrInu CLEAs showed operational stability in the batch synthesis of IFOs.
Highlights
Inulin-type fructooligosaccharides (IFOs) are well known as excellent prebiotics since they promote the growth of beneficial bacteria in the colon [1]
IFOs have been synthesized by hydrolysis of inulin using endo-inulinase [9,10]
IFOs synthesis using an immobilized our work has reported on IFOs synthesis using an immobilized inulosucrase [25] and no reports have inulosucrase reports have made the preparation
Summary
Inulin-type fructooligosaccharides (IFOs) are well known as excellent prebiotics since they promote the growth of beneficial bacteria in the colon [1]. IFOs have been used as a low-calorie sweetener [2]. They can be synthesized enzymatically from sucrose using a β-fructofuranosidase (E.C. 3.2.1.26) derived from fungal species such as Aspergillus niger [3,4], Aspergillus japonicas [5,6], Aspergillus kawachii [7], and Aspergillus oryzae [8]. IFOs have been synthesized by hydrolysis of inulin using endo-inulinase [9,10]. In comparison with fungal β-fructofuranosidase, inulosucrase has a higher transglycosylation activity [16]; it synthesizes both inulin and IFOs from sucrose, while β-fructofuranosidase mainly produces hydrolysis products and short chain oligosaccharides
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