Gene Cloning and Sequence Determination of a Novel Cyclodextrin Glycosyltransferase with a High Conversion Rate of Gamma-cyclodextrin Produced by 2 Bacillus Brevis CD162

摘要

The present invention relates to gene cloning and gene sequencing of a novel cyclodextrin glucosyltransferase produced by Bacillus brevis CD162. Cyclodextrin glycosyltransferases are enzymes that make cyclodextrins from starch. Cyclodextrins are donut-shaped cyclic oligosaccharides, which are classified as α-cyclodextrin by 6 glucose, beta-cyclodextrin by 7 glucose, and gamma-cyclodextrin by 8 glucose. The unique structure of the annulus is that in the aqueous solution, the inner surface of the cavity in the cyclodextrin molecule is hydrophobic and the outer surface is hydrophilic so as to encapsulate various organic and inorganic chemicals in the cavity (Szejtli, J. Carbo hydrate Polymers. 12: 375-392.1990). Cyclodextrins, which have been noted as molecular capsules for improving various physical properties by using these clathrates, are characterized by odour masking, taste masking and removal of bitterness, stabilization of volatiles, sweetening and stabilizing low-calorie sweeteners, and photolysis It is widely used industrially because it has the effect of protecting acid and oxides, emulsifying, stabilizing unstable substances, solubilizing poorly soluble substances, and powdering deliquescent and adhesive substances (Parrish, MAcyclodextrin-a review. 42 1393. 1989). Gamma-cyclodextrin in cyclodextrin has a large solubility due to the large size of the cavity compared to alpha- and beta-cyclodextrin, so it is very advantageous for industrial use. However, since gamma-cyclodextrin is difficult to mass-produce and the unit cost is much higher than that of alpha- and beta-cyclodextrin, there are many limitations in practical use. Therefore, the present invention cloned the gene of the cyclodextrin glycosyltransferase and revealed the sequence of the gene, which is to build a mass expression system of genes capable of producing a large amount of cyclodextrin glycosyltransferase to inexpensively mass-produce enzymes. It can be used very usefully in industries such as food, medicine, cosmetics, resin products and pesticides.