Characterization of cyclodextrin glucanotransferase produced by Bacillus megaterium.
Cyclodextrin glucanotransferase, produced by Bacillus megaterium, was characterized, and the biochemical properties of the purified enzyme were determined. The substrate specificity of the enzyme was tested with different alpha-1,4-glucans. Cyclodextrin glucanotransferase displayed maximum activity in the case of soluble starch, with a Km value of 3.4 g/L. The optimal pH and temperature values for the cyclization reaction were 7.2 and 60 degrees C, respectively. The enzyme was stable at pH 6.0-10.5 and 30 degrees C. The enzyme activity was activated by Sr2+, Mg2+, Co2+, Mn2+, and Cu2+, and it was inhibited by Zn2+ and Ag+. The molecular mass of cyclodextrin glucanotransferase was established to be 73,400 Da by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 68,200 Da by gel chromatography, and 75,000 Da by mass spectrometry. The monomer form of the enzyme was confirmed by the analysis of the N-terminal amino acid sequence. Cyclodextrin glucanotransferase formed all three types of cyclodextrins, but the predominant product was beta-cyclodextrin.
Department of Biochemistry and Molecular Biology, University of Food Technologies, 26 Maritza Blvd., 4000 Plovdiv, Bulgaria.
Ivan
I
Pishtiyski
Viara
V
Popova
Boriana
B
Zhekova
D000595:Amino Acid Sequence; D001410:Bacillus megaterium / Q000201:enzymology*; D003500:Cyclization; D003505:Cyclodextrins / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D008670:Metals / Q000378:metabolism / Q000494:pharmacology; D008970:Molecular Weight; D013379:Substrate Specificity; D013696:Temperature
D003505:Cyclodextrins; D007202:Indicators and Reagents; D008670:Metals; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase