Methods to Determine Enzymatic Activity

Methods to Determine Chitinolytic Activity

Author(s): Rosalie R. R. Coelho, Juliana P. Rosa, Rodrigo F. Souza and Adriana M. Fróes

Pp: 39-67 (29)

DOI: 10.2174/9781608053001113010007

* (Excluding Mailing and Handling)


In this chapter methods for the selection of chitinolytic microorganisms is discussed, especially those based on the formation of clear hydrolytic zones due to colloidal chitin degradation. Also the main methods for the detection and measurement of chitinolytic activity are presented, including several assays for exochitinase and endochitinase, based on artificial substrates, as well as those based on reducing sugars from chitinous substrates, such as glycol-chitin or colloidal chitin. In addition, methods for the detection of chitinase in polyacrylamide gel electrophoresis are discussed.

Keywords: Bioprospection, endochitinase, exochitinase, chitinolytic activity, colloidal chitin, glycol-chitin, chromogenic substrates, fluorogenic substrates, methylumberlliferyl, Nelson-Somogyi method, p-nitrophenyl, reducing sugars, SDS_PAGE, staining dyes, zymogram, chitinase, β-N-acetyl glucosaminidase, pre-enrichment, chitobiase, N-acetylglucosamine.

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