title: Chemical modification of lysine side chains of cyclodextrin glycosyltransferase from Thermoanaerobacter causes a shift from cyclodextrin glycosyltransferase to alpha-amylase specificity.
abstract: Cyclodextrin glycosyltransferases and alpha-amylases are two groups of enzymes with related secondary structures. However, cyclodextrin glycosyltransferases display transferase activities not present in alpha-amylases, probably derived from the existence of two more domains and different amino acid sequences. The hydrolytic activity of cyclodextrin glycosyltransferases is generally quite low, except for two cyclodextrin glycosyltransferases from termophiles. In this work, we have carried out the chemical modification (with acetic anhydride) of the amino groups of cyclodextrin glycosyltransferase from Thermoanaerobacter to assess their contributions to protein function. The acetylated cyclodextrin glycosyltransferase showed a significant reduction of its cyclization, coupling and disproportionation activities. Surprisingly, the hydrolytic (saccharifying) activity was slightly enhanced. These results suggest the participation of one or more lysine side chains in the interactions contributing to the transferase activity, either in any of the S11 subsites or in the acceptor binding site.
authors: Departamento de Biocatálisis, Instituto de Catálisis, CSIC, Madrid, Spain. malcalde@icp.csic.es
authors: M
authors: M
authors: Alcalde
authors: F J
authors: FJ
authors: Plou
authors: C
authors: C
authors: Andersen
authors: M T
authors: MT
authors: Martín
authors: S
authors: S
authors: Pedersen
authors: A
authors: A
authors: Ballesteros
mesh_terms: D000089:Acetic Anhydrides; D000107:Acetylation; D001421:Bacteria, Anaerobic / Q000201:enzymology*; D005964:Glucosyltransferases / Q000737:chemistry* / Q000378:metabolism*; D016988:Gram-Positive Asporogenous Rods, Irregular / Q000201:enzymology*; D008239:Lysine / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000737:chemistry*
chemical_list: D000089:Acetic Anhydrides; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase; D000516:alpha-Amylases; D008239:Lysine