Surawut Sangmanee, Santhana Nakapong, Rath Pichyangkura, and Kamontip Kuttiyawong

DOI: 10.14456/sjst-psu.2016.40

pp. 295 - 303

Abstract

Bacillus licheniformis RN-01 levansucrase Y246S (LsRN-Y246S) was immobilized by covalently linking onto chitosan,
Sepabead EC-EP, and Sepabead EC-HFA, beads. The stability of immobilized LsRN-Y246S was found to be the highest with
chitosan beads, retaining more than 70% activity after 13 weeks storage at 4 oC, and 68% activity after 12 hours incubation at
40°C. LsRN-Y246S immobilized on chitosan beads withstands sucrose concentrations up to 70% (w/v), retaining over 85% of
its activity, significantly better than LsRN-Y246S immobilized on others supporting matrices. LsRN-Y246S immobilized on
chitosan showed a 2.4 fold increase in activity in the presence of Mn²⁺, and gave slight protection against deactivation by
of Cu²⁺, Zn²⁺, Fe³⁺, SDS and EDTA. A maximum of 8.36 g and an average of 7.35 g LFOS yield at least up to DP 11 can be
produced from 25 g of sucrose, during five production cycles. We have demonstrated that LFOS can be effectively produced
by chitosan immobilized LsRN-Y246S and purified.