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Development and Application of Cyclodextrin Hydrolyzing Mutant Enzyme Which Hydrolyzes β- and γ-CD Selectively
Ye-Seul Koo1, Dam-Seul Ko1, Da-Woon Jeong1
1Department of Food Science and Nutrition and Center for Aging and Health Care, Hallym University , Hallymdaehak-gil 1, Chuncheon, Gangwon-do 24252, Korea.
Journal of Agricultural and Food Chemistry
|March 3, 2017
Summary
A novel enzyme, cyclodextrin glycosyl transferase (CGTase)-alpha, selectively hydrolyzes beta- and gamma-cyclodextrins. This specificity enables the purification of alpha-cyclodextrin to 99% purity from mixtures.
Area of Science:
- Biochemistry
- Enzymology
- Carbohydrate Chemistry
Background:
- Cyclodextrins (CDs) are starch derivatives produced by cyclodextrin glucanotransferase (CGTase).
- Alpha-cyclodextrin (α-CD) is valuable in the food industry as a molecular carrier and soluble fiber.
- Specific amino acids (K232, H233) in CGTase are crucial for enzyme specificity.
Purpose of the Study:
- To develop a novel enzyme for selective cyclodextrin hydrolysis.
- To utilize this enzyme for the purification of α-CD.
Main Methods:
- Site-directed mutagenesis was used to create CGTase-alpha.
- Action pattern analysis assessed CGTase-alpha's substrate specificity.
- Purification of α-CD involved enzymatic treatment and hydrophobic interaction chromatography.
Main Results:
- CGTase-alpha selectively hydrolyzed β- and γ-CD, but not α-CD.
- A method was developed to purify α-CD from a mixture using CGTase-alpha and glucoamylase.
- High purity (99%) α-CD was obtained.
Conclusions:
- CGTase-alpha exhibits selective cyclodextrin hydrolysis, offering a new tool for α-CD purification.
- Enzymatic modification of CGTase can yield enzymes with tailored specificity.
- This approach provides an efficient method for obtaining high-purity α-CD.
