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

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  • 1. Department of Chemistry, Moscow State University, USSR.
      Authors
    • Chernoglazov VM 1
    (1 author)

Analytical Biochemistry, 01 May 1989, 179(1):186-189
https://doi.org/10.1016/0003-2697(89)90222-4 PMID: 2757193 

Abstract 

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.

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Read article at publisher's site: https://doi.org/10.1016/0003-2697(89)90222-4

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