Wen Xin, Lin Huibin, Ning Yuhang, Liu Guangwen, Ren Yilin, Li Can, Zhang Chengjia, Lin Jianqun, Song Xin, Lin Jianqiang
State Key Laboratory of Microbial Technology, Shandong University, Qingdao, China.
Shandong Academy of Chinese Medicine, Jinan, China.
Front Microbiol. 2022 Apr 25;13:870168. doi: 10.3389/fmicb.2022.870168. eCollection 2022.
The NAD(P)-dependent alcohol dehydrogenase (ADH) gene was cloned from NBRC 3264 and expressed in BL21 star (DE3). The expressed enzyme was purified and the characteristics were investigated. The results showed that this ADH can convert allitol into D-allulose (D-psicose), which is the first reported enzyme with this catalytic ability. The optimum temperature and pH of this enzyme were 50°C and pH 7.0, respectively, and the enzyme showed a maximal activity in the presence of Co. At 1 mM Co and allitol concentrations of 50, 150, and 250 mM, the D-allulose yields of 97, 56, and 38%, respectively, were obtained after reaction for 4 h under optimal conditions, which were much higher than that obtained by using the epimerase method of about 30%.
从日本技术评价研究所（NBRC）3264中克隆出NAD(P)依赖型乙醇脱氢酶（ADH）基因，并在BL21 star（DE3）中表达。对表达的酶进行纯化并研究其特性。结果表明，这种ADH可以将阿洛糖醇转化为D-阿洛酮糖（D-塔格糖），这是首次报道具有这种催化能力的酶。该酶的最适温度和pH分别为50°C和pH 7.0，并且该酶在Co存在下表现出最大活性。在1 mM Co和50、150和250 mM的阿洛糖醇浓度下，在最佳条件下反应4小时后，D-阿洛酮糖的产率分别为97%、56%和38%，远高于使用差向异构酶方法获得的约30%的产率。
