谷氨酸棒杆菌ATCC14067海藻糖合酶基因的克隆及功能鉴定

发布时间：2018-06-17 08:55

  本文选题：海藻糖 + 酶法制备　； 参考：《食品科技》2017年08期

【摘要】：海藻糖分子是由2个吡喃葡萄糖环以α,α-1,1-糖苷键连接而成的一种十分稳定的非还原二糖,由于其特殊的生物学保护功用,广泛应用于医药、化妆品、食品等多个领域,具有很好的市场前景。目前,海藻糖的生产方法以生物酶法制备为主。其中,利用Tre Y-Tre Z途径由淀粉生产海藻糖和利用Tre S途径由麦芽糖生产海藻糖最为经济可行。针对谷氨酸棒杆菌ATCC14067的海藻糖合酶基因(tre S)进行基因克隆,将其插入表达质粒p ET-22b,在宿主菌E.coli BL21(DE3)中进行诱导表达,成功得到特异表达的目的蛋白。经高效液相色谱检测证实,该重组酶作用于麦芽糖后,可以产生海藻糖,具有海藻糖合酶的相应活性。在最适反应条件(30℃,p H7.5)下海藻糖合酶的转化效率可达60%~70%。实验系统地对谷氨酸棒杆菌ATCC14067的海藻糖合酶基因进行了异源表达及酶学性质的研究,为其大规模生产及工业化应用奠定了基础。
[Abstract]:Trehalose molecule is a kind of stable non-reducing disaccharide which is composed of two glucopyranosaccharide rings connected by 伪, 伪 -1ta-1-glucoside bond. Due to its special biological protective function, trehalose is widely used in many fields, such as medicine, cosmetics, food, etc. There is a good market prospect. At present, trehalose is mainly produced by biological enzyme method. Among them, trehalose production from starch by Tre Y-Tre Z pathway and trehalose from maltose by Tre S pathway is the most economical and feasible. The trehalose synthase gene of Corynebacterium glutamicum ATCC14067 was cloned and inserted into the expression plasmid pET-22b. High performance liquid chromatography (HPLC) showed that the recombinant enzyme could produce trehalose and had the corresponding activity of trehalose synthase after acting on maltose. The conversion efficiency of trehalose synthase reached 60% under the optimum reaction conditions (30 鈩，

本文编号：2030466