固定化酶两步催化合成阿糖鸟苷-微生物学专业论文.docxVIP

ABSTRACT9-beta-D·arabinofuranosyl ABSTRACT 9-beta-D·arabinofuranosyl guanine(ara-G)，an active metabolite of nelarabine，is kind of drug which has anti—leukemia effects．Am—G Can be phosphorylated to ara-G triphosphate(ara·GTP)in the human body。 Am—GTP competes with deoxynucleoside triphosphate to insert into DNA．And then it to selectively suppress the synlhesis of DNA．The chemical synthesis of ara—G is extremely complex,bringing about different kinds of isomer and hard to separate from isomer．Besides if using free cells enzymes for biosynthesis，such one-off using will cost too much．In this paper,immobilized enzyme technology was used in the biosynthetic research of aiR-GA continuous catalytic reaction process will be established，which tremendously reduced the production cost．This research provide the reference for the industrialized production of ara—G In the preparation of immobilized enzyme，enzyme solution was obtained by using ultrasonic， ammonium sulfate fractionation．Immobilized enzyme which be useA for catalytic conversion reaction was obtained by fixing resin carrier with glutaraldehyde crosslinking method．The immobilized conditions of nucleoside phosphorylase were optimized following：the materials were set in 25。C，30 rpm and mixed with 0．2％glutaraldehyde for 4h，63．2 enzyme fixed lg resin carrier,and the enzyme linked with resin lasting for 1 2h．Then resin were washed by distilled water tO the neutral pH and preserved in the 25 mmol／L pH 7 PBS buffer．The immobilized conditions of adenosine deaminase were optimized following：the materials were set in 25。(2，30 rpm and mixed with 0．1％glutaraldehyde for 8h，5 1．4 enzyme fixed l g resin carrier,and the enzyme linked with resin lasting for 1 0h．Then resin were washed by distilled water to the neutral pH and preserved in the 25 mmoFL pH 7 PBS buffer． III 万方数据 At At first，biosynthesis system of ara-DA was built：the substrates ara-U and DAP Were dissolved in phosphate buffer,using immobilized purine nucleoside phosphor