Qingdao Institute of Bioenergy and Bioprocess Technology and Key Laboratory of Bioenergy, Chinese Academy of Sciences, Qingdao, 266101, China.
Biochemistry (Mosc). 2013 Apr;78(4):424-30. doi: 10.1134/S0006297913040123.
By combining anion-exchange chromatography with gel filtration, an effective method for purification of wild-type xyloglucanase and five other cellulolytic enzymes from strain QM9414 of Trichoderma reesei was established. Characterization by enzyme activity assay, SDS-PAGE, and mass spectrometry identified the purified proteins as cellobiohydrolases I and II, endoglucanases I and II, a xyloglucanase, and β-xylosidase, of which the xyloglucanase was purified for the first time from the mutant strain QM9414. This method holds great promise to study the mechanism of cellulolytic enzymes, to investigate the synergistic action between cellulase and other cellulolytic enzymes, and to better exploit enzyme preparations for degradation of lignocellulose.
通过阴离子交换色谱与凝胶过滤相结合，建立了一种从里氏木霉 QM9414 菌株中有效纯化野生型木聚糖酶和其他五种纤维素酶的方法。通过酶活性测定、SDS-PAGE 和质谱分析对纯化蛋白进行鉴定，确定其为纤维二糖水解酶 I 和 II、内切葡聚糖酶 I 和 II、木聚糖酶和β-木糖苷酶，其中木聚糖酶是首次从突变株 QM9414 中纯化得到的。该方法有望用于研究纤维素酶的作用机制、探究纤维素酶与其他纤维素酶之间的协同作用，以及更好地利用酶制剂来降解木质纤维素。
