Enzymatic synthesis of dimaltosyl-{beta}-cyclodextrin via a transglycosylation reaction using TreX, a Sulfolobus solfataricus P2 debranching enzyme
- Center for Agricultural Biomaterials, Department of Food Science and Biotechnology, College of Agriculture and Life Sciences, Seoul National University, San 56-1, Shillim-dong, Kwanak-gu, Seoul 151-921 (Korea, Republic of)
- Department of Food and Biotechnology, Korea University, Jochiwon, Chungnam 339-700 (Korea, Republic of)
- Department of Food Science and Technology, Chungang University, Anseong 456-756 (Korea, Republic of)
- School of Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya 663-8179 (Japan)
- Department of Biology, University of Incheon, Incheon 402-749 (Korea, Republic of)
- Department of Biological Science, Pusan National University, Busan 609-735 (Korea, Republic of)
- Department of Food and Nutrition, Seowon University, Cheongju 361-742 (Korea, Republic of)
Di-O-{alpha}-maltosyl-{beta}-cyclodextrin ((G2){sub 2}-{beta}-CD) was synthesized from 6-O-{alpha}-maltosyl-{beta}-cyclodextrin (G2-{beta}-CD) via a transglycosylation reaction catalyzed by TreX, a debranching enzyme from Sulfolobus solfataricus P2. TreX showed no activity toward glucosyl-{beta}-CD, but a transfer product (1) was detected when the enzyme was incubated with maltosyl-{beta}-CD, indicating specificity for a branched glucosyl chain bigger than DP2. Analysis of the structure of the transfer product (1) using MALDI-TOF/MS and isoamylase or glucoamylase treatment revealed it to be dimaltosyl-{beta}-CD, suggesting that TreX transferred the maltosyl residue of a G2-{beta}-CD to another molecule of G2-{beta}-CD by forming an {alpha}-1,6-glucosidic linkage. When [{sup 14}C]-maltose and maltosyl-{beta}-CD were reacted with the enzyme, the radiogram showed no labeled dimaltosyl-{beta}-CD; no condensation product between the two substrates was detected, indicating that the synthesis of dimaltosyl-{beta}-CD occurred exclusively via transglycosylation of an {alpha}-1,6-glucosidic linkage. Based on the HPLC elution profile, the transfer product (1) was identified to be isomers of 6{sup 1},6{sup 3}- and 6{sup 1},6{sup 4}-dimaltosyl-{beta}-CD. Inhibition studies with {beta}-CD on the transglycosylation activity revealed that {beta}-CD was a mixed-type inhibitor, with a K{sub i} value of 55.6 {mu}mol/mL. Thus, dimaltosyl-{beta}-CD can be more efficiently synthesized by a transglycosylation reaction with TreX in the absence of {beta}-CD. Our findings suggest that the high yield of (G2){sub 2}-{beta}-CD from G2-{beta}-CD was based on both the transglycosylation action mode and elimination of the inhibitory effect of {beta}-CD.
- OSTI ID:
- 21043599
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 366, Issue 1; Other Information: DOI: 10.1016/j.bbrc.2007.11.115; PII: S0006-291X(07)02507-7; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
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