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Production of 2-O-α-glucopyranosyl l-ascorbic acid from ascorbic acid and β-cyclodextrin using immobilized cyclodextrin glycosyltransferase

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Abstract

Cyclodextrin glycosyltransferase (CGTase) isolated and purified from Paenibacillus sp. A11 was immobilized on various carriers by covalent linkage using bifunctional agent glutaraldehyde. Among tested carriers, alumina proved to be the best carrier for immobilization. The effects of several parameters on the activation of the support and on the immobilization of enzyme were optimized. The best preparation of immobilized CGTase retained 31.2% of its original activity. After immobilization, the enzymatic properties were investigated and compared with those of the free enzyme. The optimum pH of the immobilized CGTase was shifted from 6.0 to 7.0 whereas optimum temperature remained unaltered (60°C). Free and immobilized CGTase showed similar pH stability profile but the thermal stability of the immobilized CGTase was 20% higher. Kinetic data (K M and V max) for the free and immobilized enzymes were determined from the rate of β-CD formation and it was found that the immobilized form had higher K M and lower V max. The immobilized CGTase also exhibited higher stability when stored at both 4°C and 25°C for 2 months. The enzyme immobilized on alumina was further used in a batch production of 2-O-α-glucopyranosyl-l-ascorbic acid (AA-2G) from ascorbic acid and β-cyclodextrin. The yield of AA-2G was 2.92% and the immobilized CGTase retained its activity up to 74.4% of the initial catalytic activity after being used for 3 cycles. The immobilized CGTase would have a promising application in the production of various transglycosylated compounds and in the production of cyclodextrin by the hydrolysis of starch.

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Abbreviations

AA:

Ascorbic acid

AA-2G:

2-O-α-glucopyranosyl-l-ascorbic acid

AANa:

l-Ascorbic acid sodium salt

APTS:

Aminopropyltriethoxysilane

CD:

Cyclodextrin

CGTase:

Cyclodextrin glycosyltransferase

GA:

Glutaraldehyde

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Acknowledgments

This work was supported by a grant from Starch and Cyclodextrin Unit Cell, Chulalongkorn University (Bangkok, Thailand). We are grateful to Dr. Rath Pichayankura for providing chitosan used in this study.

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  1. Department of Biochemistry, Faculty of Science, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand

    Manchumas Hengsakul Prousoontorn & Supranee Pantatan

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  1. Manchumas Hengsakul Prousoontorn
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  2. Supranee Pantatan
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Correspondence to Manchumas Hengsakul Prousoontorn.

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Prousoontorn, M.H., Pantatan, S. Production of 2-O-α-glucopyranosyl l-ascorbic acid from ascorbic acid and β-cyclodextrin using immobilized cyclodextrin glycosyltransferase. J Incl Phenom Macrocycl Chem 57, 39–46 (2007). https://doi.org/10.1007/s10847-006-9163-5

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  • DOI: https://doi.org/10.1007/s10847-006-9163-5

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