Tomaz Candida T, Duarte Dina, Queiroz João A
Departamento de Química, Universidade do Beira Interior, Covilhã, Portugal.
J Chromatogr A. 2002 Jan 25;944(1-2):211-6. doi: 10.1016/s0021-9673(01)01224-9.
In this work, a comparative study for the fractionation of Trichoderma reesei cellulases on five different hydrophobic interaction chromatography adsorbents (Butyl-Sepharose 4 FF, Phenyl-Sepharose 6 FF, Octyl-Sepharose 4 FF, Epoxy-Sepharose CL-6B and Polypropylene glycol-Sepharose CL-6B) is shown. The influence of the mobile phase composition on the chromatographic behaviour of T. reesei cellulases complex was evaluated using different concentrations of ammonium sulphate in the eluent buffer. A selective separation of beta-glucosidase with two-fold increase in specific activity and good recoveries of cellobiase activity were obtained with Butyl-Sepharose 4 FF and Phenyl-Sepharose 6 FF using 7% (w/v) ammonium sulphate in the eluent buffer. A beta-glucosidase fractionation was also obtained with Epoxy-Sepharose CL-6B, using 13% (w/v) of the salt in the mobile phase.
本研究展示了在五种不同的疏水相互作用色谱吸附剂（丁基琼脂糖凝胶4 FF、苯基琼脂糖凝胶6 FF、辛基琼脂糖凝胶4 FF、环氧琼脂糖凝胶CL - 6B和聚丙二醇琼脂糖凝胶CL - 6B）上对里氏木霉纤维素酶进行分级分离的比较研究。使用洗脱缓冲液中不同浓度的硫酸铵，评估了流动相组成对里氏木霉纤维素酶复合物色谱行为的影响。使用洗脱缓冲液中7%（w/v）的硫酸铵，在丁基琼脂糖凝胶4 FF和苯基琼脂糖凝胶6 FF上获得了β - 葡萄糖苷酶的选择性分离，其比活性提高了两倍，纤维二糖酶活性的回收率良好。使用流动相中13%（w/v）的盐，在环氧琼脂糖凝胶CL - 6B上也实现了β - 葡萄糖苷酶的分级分离。
