Continuous photometric determination of endo-1,4-β-d-glucanase (cellulase) activity using 4-methylumbelliferyl-β-d-cellobioside as a substrate

✍ Scribed by Vladimir M. Chernoglazov; Aida N. Jafarova; Anatole A. Klyosov

• Publisher: Elsevier Science
• Year: 1989
• Tongue: English
• Weight: 430 KB
• Volume: 179
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(89)90222-4

✦ Synopsis

A very simple and sensitive procedure for the determination of the activity of highly purified endo-1,4-beta-glucanase from the microscopic fungus Trichoderma reesei using 4-methylumbelliferyl-beta-D-cellobioside has been developed. The HPLC study has shown that this substrate is cleaved by endo-1,4-beta-glucanase to form predominantly free 4-methylumbelliferone, Km and kcat being 1.25 mM and 7.9 s-1, respectively (30 degrees C, pH 5.0). The possibility of continuous photometric determination of the enzyme using the difference absorptivity coefficient of 1600 M-1 cm-1 at 350 nm has been demonstrated.