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Invention Publication
CN104073476A 一种麦芽糊精底物特异性提高的环糊精糖基转移酶及其制备方法 有权

一种麦芽糊精底物特异性提高的环糊精糖基转移酶及其制备方法
Abstract:
本发明公开了一种麦芽糊精底物特异性提高的环糊精糖基转移酶及其制备方法，属于遗传工程和酶工程领域。本发明通过对P.macerans strain JFB05-01(CCTCC NO:M208063)的CGTase的195位的酪氨酸(Tyr)替换为丝氨酸(Ser),260位的酪氨酸(Tyr)替换为精氨酸(Arg),265位的谷氨酰胺(Gln)替换为赖氨酸(Lys)，使AA-2G产量分别提高了23％，44％和40％。对以上突变株组合突变，获得双突变体Y195S/Y260R,Y195S/Q265K和Y260R/Q265K以及三点突变体Y195S/Y260R/Q265K。它们利用麦芽糊精为糖基供体生产AA-2G产量分别提高了57％，49％，55％和59％。这些突变株比野生型CGTase更利于利用麦芽糊精为糖基供体生产AA-2G。
Abstract(English):

The invention discloses a cyclodextrin glycosyltransferase with maltodextrin substrate specificity being improved and a preparation method thereof, belonging to the field of genetic engineering and enzyme engineering. By replacing 195-site Tyr of CGTase of a P.macerans strain JFB05-01 (CCTCC NO:M208063) with Ser, 260-site Tyr with Arg and 265-site Gln with Lys, the output of AA-2G can be respectively increased by 23%, 44% and 40%. Combination mutation is conducted to the mutant strains, double mutants Y195S/Y260R, Y195S/Q265K and Y260R/Q265K and three-point mutants Y195S/Y260R/Q265K can be obtained. The maltodextrin is utilized for producing AA-2G for a glycosyl donor, the output of the AA-2G is respectively improved 57%, 49%, 55% and 59%. The mutant strains are more favorable to produce AA-2G for the glycosyl donor by utilizing maltodextrin compared with the wild CGTase.

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