Production of cyclodextrin by poly-lysine fused Bacillus macerans cyclodextrin glycosyltransferase immobilized on cation exchanger

Bacillus macerans cyclodextrin glycosyltransferase (CGTase) fused with 10 lysine residues at its C-terminus (CGTK10ase) was immobilized onto a cation exchanger by ionic interaction and used to produce α-cyclodextrin (CD) from soluble starch. Poly-lysine fused immobilization increased the Vm of the immobilized CGTase by 40% without a change in Km. The activation energies of thermal deactivation (Ea) were 41.4, 28.1, and 25.9 kcal mol−1, respectively, for soluble wild-type (WT) CGTase, soluble CGTK10ase, and immobilized CGTK10ase, suggesting destabilization of CGTase by poly-lysine fusion and immobilization onto a cation exchanger. Maximum α-CD productivity of 539.4 g l−1 h−1 was obtained with 2% soluble starch solution which was constantly fed at a flow rate of 4.0 ml min−1 (D = 240 h−1) in a continuous operation mode of a packed-bed reactor. The operational half-life of the packed-bed enzyme reactor was estimated 12 days at 25 °C and pH 6.0.