| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 70326 | Journal of Molecular Catalysis B: Enzymatic | 2011 | 7 Pages |
Bacillus macerans cyclodextrin glycosyltransferase (CGTase) (EC 2.4.1.19) was covalently immobilised on Eupergit C and used in a packed-bed reactor to investigate the continuous production of long-carbohydrate-chain alkyl glycosides from α-cyclodextrin (α-CD) and n-dodecyl-(1,4)-β-maltopyranoside (C12G2β). The effects of buffer ion strength and pH, and enzyme loading on the immobilisation yield and the enzyme activity were evaluated. Approximately 98% of the protein and 33% of the total activity were immobilised. At pH 5.15, the enzymatic half-life was 132 min at 60 °C and 18 min at 70 °C. The immobilised enzyme maintained 60% of its initial activity after 28 days storage at 4 °C. The degree of conversion was controlled by simple regulation of the flow rate through the reactor, making it possible to optimise the product distribution. It was possible to achieve a yield of the primary coupling product n-dodecyl-(1,4)-β-maltooctaoside (C12G8β) of about 50%, with a ratio between the primary and the secondary coupling product of about 10. Thermoanaerobacter sp. CGTase (Toruzyme 3.0 L) immobilised on Eupergit C had good operational stability at 60 and 70 °C thus showing the advantages of using more thermostable enzymes in biocatalysis. However, this enzyme was unsuitable for the production of C12G8β due to extensive disproportionation reactions, giving a broad product range.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideResearch highlights► Immobilization of two cyclodextrin glycosyltransferases on Eupergit C. ► Conversion of dodecyl-β-maltoside and α-cyclodextrin into dodecyl-β-maltooctaoside. ► Side reactions were minimized by flow rate control of a packed bed reactor.
