Bartley T D, Murphy-Holland K, Eveleigh D E
Anal Biochem. 1984 Jul;140(1):157-61. doi: 10.1016/0003-2697(84)90147-7.
Endoglucanase and exoglucanase components of cellulase can be detected and differentiated after polyacrylamide gel electrophoresis by performing activity stains. Endoglucanase activity was visualized in carboxymethyl cellulose agar replicas of gels by staining with Congo red. General beta-1,4-glucanase activity was located by soaking the gel in a solution of NaBH4-reduced cellulooligosaccharides, and detecting the formation of reducing sugars by reaction with triphenyl tetrazolium chloride. Endoglucanases are active in both assays, while exoglucanases can be distinguished by their activity in the cellulo-oligosaccharide assay only. This methodology has facilitated the purification and characterization of cellulase components from Trichoderma reesei and Microbispora bispora.
纤维素酶的内切葡聚糖酶和外切葡聚糖酶组分在聚丙烯酰胺凝胶电泳后,通过活性染色可以被检测和区分。内切葡聚糖酶活性通过在凝胶的羧甲基纤维素琼脂复制品上用刚果红染色来可视化。通用的β-1,4-葡聚糖酶活性通过将凝胶浸泡在硼氢化钠还原的纤维寡糖溶液中,并通过与氯化三苯基四氮唑反应检测还原糖的形成来定位。内切葡聚糖酶在两种测定中均有活性,而外切葡聚糖酶只能通过其在纤维寡糖测定中的活性来区分。这种方法促进了里氏木霉和双孢小双孢菌纤维素酶组分的纯化和表征。