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A new spectrophotometric method of assay for chitosanase based on Calcofluor white dye binding.

Somashekar, D. and Joseph, R. (1997) A new spectrophotometric method of assay for chitosanase based on Calcofluor white dye binding. Carbohydrate Polymers, 34 (4). 343-346, 15 ref..

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Abstract

A new spectrophotometric method for the assay of chitosanase based on complex formation of the substrate chitosan with Calcofluor white dye is described. The absorption maximum for the chitosan--Calcofluor complex is determined to be 406 nm. The apparent minimum size of chitosan for complex formation is 5-7 kDa. Therefore, those enzymes that do not generate glucosamine or reducing groups as products of hydrolysis at levels not measurable by the available methods of assay can be assayed by the present method. In the standardized procedure 2OOpg of chitosan in acetate buffer pH 4.5 with the enzyme in a reaction volume of 1 S ml is incubated at 45°C for 1 h, after which 1.5 ml of Calcofluor white (0.05%) is added, kept for 1 h and absorbance at 406nm measured by a spcctrophotometer. The chitosanase unit is arbitrarily defined as the reduction in absorbance by O.Ol/min.

Item Type: Article
Uncontrolled Keywords: Chitosanases spectrophotometric method Calcofluor white dye
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology
500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 16 Enzyme Chemistry
Divisions: Food Microbiology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 20 Nov 2008 10:12
Last Modified: 28 Dec 2011 09:40
URI: http://ir.cftri.com/id/eprint/2498

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