| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The gene of inulosucrase from Lactobacillus reuteri 121 (inu; GenBank accession number AF459437) was synthesized by Genscript. The synthetic gene was subcloned into pET21-b via NdeI and XhoI sites. The recombinant vector (pETIns) was transformed into Escherichia coli BL21 (DE3). Plasmid-carrying E. coli strains were grown at 30 °C in the Luria-Bertani medium, supplemented with 100 μg mL−1 ampicillin, 10 mM CaCl2, and 0.1 mM IPTG for enzyme induction. After 18 h, the cells were harvested using centrifugation (5000 × g) at 4 °C for 10 min. The cell pellet was resuspended in 50 mM sodium citrate buffer pH 5.0 and then sonicated. Cell debris were removed by centrifugation (10 000 × g, 4 °C, 15 min) to obtain the crude extract of the enzyme. | Get A Quote |
With the aim to overcome the limitations of hydrogel chitosan beads (HGBs), various types of chitosan, core–shell chitosan beads (CSBs), and dried chitosan beads (DBs) were synthesized. Physical and chemical properties were compared with those of HGBs. CSBs were proved to be an effective support because they displayed higher stability and capacity over the HGBs, and thus, were selected for enzyme immobilization. Recombinant inulosucrase (INU) from Lactobacillus reuteri 121 was immobilized on CSBs using glutaraldehyde as a cross-linker. Immobilized biocatalysts (INU-CSBs) were then used for the synthesis of inulin-type fructooligosaccharide (IFOS). Biochemical characterization revealed that the optimum pH of... More
