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Updated: Apr 4, 2026
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
Published on: October 4, 2017
Development and characterization of cyclodextrin glucanotransferase as a maltoheptaose-producing enzyme using
Ye-Seul Koo1, Hye-Won Lee1, Hye-Yeon Jeon1
1Department of Food Science and Nutrition, Hallym University, Chuncheon, Republic of Korea.
Abstract:
Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) mainly produces cyclodextrins (CDs) using linear maltooligosaccharides. We performed site-directed saturation mutagenesis on the +1 substrate-binding residue, H233, of CGTase from alkalophilic Bacillus sp. I-5 to prepare specific-length oligosaccharides. The obtained mutant CGTase, H233Y, primarily produced maltoheptaose (G7) using β-CD via a hydrolysis reaction. A kinetic study of H233Y showed that the kcat/Km value of β-CD was 7-fold greater than that of G7, which accounts for the accumulation of G7 during the H233Y enzyme reaction. A structure comparison of CGTases with H233Y modeling suggests that the substitution of H233Y may alter the position of the +1 subsite and slow the further hydrolysis of G7 after the ring-opening reaction.
