A rapid and sensitive method for screening of chitinase inhibitors using Ostazin Brilliant Red labelled chitin as a substrate for chitinase assay.

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Shirazi F, Kulkarni M, Deshpande MV

A rapid and sensitive method for screening of chitinase inhibitors using Ostazin Brilliant Red labelled chitin as a substrate for chitinase assay.

Lett Appl Microbiol. 2007 Jun;44(6):660-5.

PubMed ID

17576230 [ View in PubMed

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Abstract

AIMS: The development of a simple, rapid and sensitive method for the screening of chitinase inhibitors using Ostazin Brilliant Red labelled chitin as a substrate for chitinase assay. METHODS AND RESULTS: The colloidal substrate prepared from the Ostazin Brilliant Red labelled chitin was used in the estimation of chitinase activity with and without the addition of potential chitinase inhibitor. The dye labelled N-acetylglucosamine (GlcNAc) released because of hydrolysis of the substrate was measured spectrophotometrically at 530 nm. The release of dye labelled GlcNAc from the colloidal Red chitin was proportional to the chitinase activity measured using other methods. The activities measured using acid swollen chitin and -Red chitin were comparable in the range 0.1-0.5 U ml(-1). The addition of chitinase inhibitor(s) (1000 ppm) proportionally reduced the release of dye labelled GlcNAc from the substrate in the reaction mixture. CONCLUSIONS: The assay for chitinase activity measuring the release of dye-labelled GlcNAc was simple, rapid and sensitive. There was no need to measure GlcNAc using a separate colorimetric method. SIGNIFICANCE AND IMPACT OF THE STUDY: The method provides a simple, one-step procedure for quantitative estimation of total chitinolytic activity and for screening of endochitinase and/or N-acetylglucosaminidase inhibitors acting singly or in combination. The method has a great potential in developing medium throughput procedure for the screening of chitinase inhibitors using 96-well plate reader.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
N-Acetylglucosamine	Alpha-N-acetylglucosaminidase	Protein	Humans	Unknown	Activator	Details