Recombinant Cyclodextrinase from Thermococcus kodakarensis KOD1: Expression, Purification, and Enzymatic Characterization

Table 1

Comparison of the biochemical properties of CDase-Tk and those of other CDases from archaea.

	CDase-Tk	CDase-Tc	CDase-Pf					CDase-Tb	CDase-Tp
Homology	100%	59%	56%					53%	52%
aa residues	656	644	645					660	644
Optimal temperature	65	85	90					95	95
Optimal pH	7.5	5.0	4.5					5.5	5.5
Optimal substrate	β-CD	α-CD	α-CD					β-CD	γ-CD
Substrate preference	CD ≫ PL > SS	CD ≫ MD > SS > PL	CD ≫ MD > SS					CD ≫ MD > SS	CD ≫ MD > PL = SS
Final hydrolysis product	G1	G1, G2	G3, G4					G1, G2	G1, G2
(mg mL^−1)	3.1	N.D.	-CD	-CD	-CD	MD	SS	N.D.	N.D.
			2.6	2.2	5.1	62.9	0.5
(s^−1)	34.6	N.D.	-CD	-CD	-CD	MD	SS	N.D.	N.D.
			241	196	173	268	67
/	11.1	N.D.	-CD	-CD	-CD	MD	SS	N.D.	N.D.
			92.3	90.7	33.8	4.3	128.8
References	This study	[10]	[12]					[9]	[11]

MD, maltodextrin; CA, cycloamylose; CD, cyclodextrin; PL, pullulan; SS, soluble starch.
The cyclodextrinases are from T. kodakarensis KOD1 (CDase-Tk), Thermococcus sp. CL1 (CDase-Tc), P. furiosus (CDase-Pf), Thermococcus sp. B1001 (CDase-Tb), and Thermofilum pendens Hrk 5 (CDase-Tp). N.D.:not determined.