Preparation of Cross-Linked Enzyme Aggregates (CLEAs) of an Inulosucrase Mutant for the Enzymatic Synthesis of Inulin-Type Fructooligosaccharides

Catalysts

Rath Pichyangkura¹  ManchumasHengsakul Prousoontorn¹  Thanapon Charoenwongpaiboon¹  Robert A. Field² 

[1] Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Bangkok 10330, Thailand;Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK;

关键词: inulosucrase;    cross-linked enzyme aggregates;    fructooilgosaccharide;   

DOI  :  10.3390/catal9080641

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown