| 热稳定6-磷酸-β-葡萄糖苷酶TteBglB异源表达、分离纯化及酶学性质分析 |
| |
| 引用本文： | 刘晴,§,张宇微§,张宇宏,徐欣欣,张伟,刘波,顾青.热稳定6-磷酸-β-葡萄糖苷酶TteBglB异源表达、分离纯化及酶学性质分析[J].中国农业科技导报,2014,16(6):52-58. |
| |
| 作者姓名： | 刘晴 § 张宇微§ 张宇宏 徐欣欣 张伟 刘波 顾青 |
| |
| 作者单位： | (1.浙江工商大学食品与生物工程学院, 杭州 310018,2.中国农业科学院生物技术研究所, 北京 100081) |
| |
| 基金项目： | 国家863计划项目（2012AA022105）；国家自然科学基金项目（31200072）资助。 |
| |
| 摘 要： | 6-磷酸-β-葡萄糖苷酶(EC 3.2.1.86)催化6-磷酸-葡萄糖苷类化合物(如6-磷酸-纤维二糖、6-磷酸-纤维素寡糖)产生6-磷酸-葡萄糖而使纤维素完全分解,在微生物碳源利用过程中起重要作用。腾冲嗜热厌氧杆菌是一株嗜热厌氧微生物,提供了丰富的热稳定性蛋白基因资源。从该菌株中克隆编码6-磷酸-β-葡萄糖苷酶的基因Ttebgl B,并在E.coli BL21(DE3)进行了异源表达。结果表明,TteBglB催化反应的最适p H为6.0、最适温度为70℃,在pH 4~10或70℃时有着良好的稳定性;同时证实,TteBglB属于糖基水解酶家族1(GH1)成员,可不依赖Mn2+、Ni2+、Co2+或Fe2+等二价金属离子而发挥催化作用。以pNPβG6P为底物时,催化反应的Km为0.054 mmol/L,Kcat为81.47/min,Vmax为0.003992 mmol/min,酶比活为18.093 U/mg。
|
| 关 键 词： | TteBglB 6-磷酸-β-葡萄糖苷酶 糖苷水解酶 |
Heterologous Expression,Purification and Characterization of Thermo-stable 6-phosphate-β-glucosidase TteBglB |
| |
| LIU Qing,§,ZHANG Yu-wei§,ZHANG Yu-hong,XU Xin-xin,ZHANG Wei,LIU Bo.Heterologous Expression,Purification and Characterization of Thermo-stable 6-phosphate-β-glucosidase TteBglB[J].Journal of Agricultural Science and Technology,2014,16(6):52-58. |
| |
| Authors: | LIU Qing § ZHANG Yu-wei§ ZHANG Yu-hong XU Xin-xin ZHANG Wei LIU Bo |
| |
| Institution: | (1.College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018|2.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
| |
| Abstract: | 6-phosphate-β-glucosidase (EC 3.2.1.86), an enzyme catalyzes 6-phosphate-glucoside compounds (such as 6-phosphate-cellobiose, 6-phosphate-cellulose oligosaccharides) to produce 6-phosphate-glucose, plays an important role in carbon utilization for microbes. Thermoanaerobacter tengcongensis MB4, a strain of thermophilic anaerobic microorganism, is an excellent resource for thermo-stable protein. In this study, gene TtebglB was isolated from this strain and then heterologously expressed in E.coli BL21 (DE3). The optimum pH and temperature of TteBglB were pH 6.0 and 70℃, respectively, and the enzyme kept stable at a pH 4~10 at 70℃. Additionally, it was also confirmed that TteBglB belonged to glycosyl hydrolase family 1 (GH1) and could act without Mn2+, Ni2+, Co2+, Fe2+ and other divalent metal ions. Using pNPβG6P as substrate, the values of Km, Kcat, and Vmax for this enzyme was 0.054 mmol/L, 81.47/min and 0.003 992 mmol/min, respectively, and specific activity of the enzyme reached to 18093 U/mg. |
| |
| Keywords: | TteBglB 6-phosphate-β-glucosidase glycosyl hydrolase |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国农业科技导报》浏览原始摘要信息 |
| 点击此处可从《中国农业科技导报》下载免费的PDF全文 |
|
