Rosseto Flávio R, Puhl Ana C, Andrade Maxuel O, Polikarpov Igor
Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Saocarlense 400, São Carlos-SP 13560-970, Brazil.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Feb 1;69(Pt 2):137-40. doi: 10.1107/S1744309112051408. Epub 2013 Jan 31.
Cellulases, such as endoglucanases, exoglucanases and β-glucosidases, are important enzymes used in the process of enzymatic hydrolysis of plant biomass. The bacteria Xanthomonas campestris pv. campestris expresses a large number of hydrolases and the major endoglucanase (XccEG), a member of glycoside hydrolase family 5 (GH5), is the most strongly secreted extracellularly. In this work, the native XccEG was purified from the extracellular extract and crystallization assays were performed on its catalytic domain. A complete data set was collected on an in-house X-ray source. The crystal diffracted to 2.7 Å resolution and belonged to space group C2, with unit-cell parameters a = 174.66, b = 141.53, c = 108.00 Å, β = 110.49°. The Matthews coefficient suggests a solvent content of 70.1% and the presence of four protein subunits in the asymmetric unit.
纤维素酶，如内切葡聚糖酶、外切葡聚糖酶和β-葡萄糖苷酶，是植物生物质酶解过程中使用的重要酶。野油菜黄单胞菌野油菜致病变种表达大量水解酶，主要的内切葡聚糖酶（XccEG）是糖苷水解酶家族5（GH5）的成员，是细胞外分泌最强的酶。在这项工作中，从细胞外提取物中纯化了天然XccEG，并对其催化结构域进行了结晶试验。在内部X射线源上收集了完整的数据集。晶体衍射分辨率为2.7 Å，属于空间群C2，晶胞参数a = 174.66、b = 141.53、c = 108.00 Å，β = 110.49°。马修斯系数表明溶剂含量为70.1%，不对称单元中存在四个蛋白质亚基。
