Endochitinase activity determination using N-fluorescein-labeled chitin.
A fluorimetric method for the determination of endochitinolytic activity using N-fluorescein-labeled chitin (FITC-Chitin) is proposed, and a procedure for FITC-Chitin preparation with a degree of FITC content of 2.2 mol% (one FITC molecule per 45 glucosamine residues) is described. FITC-Chitin is capable to distinguish endochitinase and exochitinase (beta-N-acetylglucosaminidase) activities.
Tikhonov
Vladimir E
VE
A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Vavilov Street, 28, 119991 Moscow, Russia. tikhon@ineos.ac.ru
Lopez-Llorca
Luis V
LV
Salinas
Jesús
J
Monfort
Elena
E
D000118:Acetylglucosaminidase / Q000378:metabolism; D001671:Biochemistry / Q000379:methods*; D002686:Chitin / Q000737:chemistry*; D002688:Chitinases / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D019793:Fluorescein / Q000737:chemistry*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry; D006596:Hexosaminidases / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D020172:Verticillium / Q000378:metabolism
D005656:Fungal Proteins; D002686:Chitin; C079639:1,4-beta-chitobiosidase; D006596:Hexosaminidases; D002688:Chitinases; D000118:Acetylglucosaminidase; D016650:Fluorescein-5-isothiocyanate; D019793:Fluorescein