State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
World J Microbiol Biotechnol. 2012 Dec;28(12):3337-44. doi: 10.1007/s11274-012-1145-8. Epub 2012 Aug 23.
By constructing the genomic library, a β-glucosidase gene, with a length of 2,382 bp, encoding 793 amino acids, designated bgla, is cloned from a marine bacterium Aeromonas sp. HC11e-3. The enzyme is expressed successfully in the recombinant host Escherichia coli BL21 (DE3) and purified using glutathione affinity purification system. It shows the optimal activity at pH 6, 55 °C and hydrolyzes aryl-glucoside specially. Ca(2+), Mn(2+), Zn(2+), Ba(2+), Pb(2+), Sr(2+) can activate the enzyme activity, whereas SDS, EDTA, DTT show slight inhibition to the enzyme activity. Homologous comparing shows that the enzyme belongs to glycosyl hydrolase family 3, exhibiting 46 % identity with a fully characterized glucosidase from Thermotoga neapolitana DSM 4359. Such results provide useful references for investigating other glucosidases in the glycosyl family 3 as well as developing glucosidases using in suitable industrial area.
通过构建基因组文库，从海洋细菌气单胞菌 HC11e-3 中克隆出一个长度为 2382bp、编码 793 个氨基酸的β-葡萄糖苷酶基因，命名为 bgla。该酶在重组宿主大肠杆菌 BL21（DE3）中成功表达，并使用谷胱甘肽亲和纯化系统进行纯化。它在 pH6、55°C 下表现出最佳活性，专门水解芳基葡萄糖苷。Ca(2+)、Mn(2+)、Zn(2+)、Ba(2+)、Pb(2+)、Sr(2+)可以激活酶活性，而 SDS、EDTA、DTT 对酶活性有轻微抑制作用。同源性比较表明，该酶属于糖苷水解酶家族 3，与来自嗜热栖热菌 DSM 4359 的完全表征的葡萄糖苷酶具有 46%的同一性。这些结果为研究糖苷家族 3 中的其他葡萄糖苷酶以及在合适的工业领域开发葡萄糖苷酶提供了有用的参考。
