Colloidal chitin stained with Remazol Brilliant Blue R®, a useful substrate to select chitinolytic microorganisms and to evaluate chitinases

M. Gómez Ramírez, L. I. Rojas Avelizapa, N. G. Rojas Avelizapa, R. Cruz Camarillo

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115 Scopus citations

Abstract

A simple and sensitive method based on the use of colloidal chitin stained with Remazol Brilliant Blue R® (RBB) is proposed to evaluate chitinase activity. If this colloidal-stained substrate is included as a carbon source in a liquid medium, this technique allows the selection or the comparison of chitinolytic microorganisms. The colloidal substrate is proportionally solubilized and the dye released is spectrophotometrically quantified at 595 nm. The procedures used for the staining and fixing of RBB in the colloidal chitin, and a comparison with the commercial substrate chitin-azure®, are presented. The influence of several physicochemical and enzymatic parameters on the release of dyes is also shown. Both stained substrates were used for studying the effect of pH, substrate concentration, temperature and time on the chitinase reaction of Bacillus thuringiensis Bt-107.

Original languageEnglish
Pages (from-to)213-219
Number of pages7
JournalJournal of Microbiological Methods
Volume56
Issue number2
DOIs
StatePublished - Feb 2004

Keywords

  • Bacillus thuringiensis
  • Chitin
  • Chitinase
  • Dyed colloidal chitin
  • Remazol Brilliant Blue-R®
  • Serratia marcescens

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