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Bioresour Technol ; 101(23): 9236-40, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20659797

RESUMEN

Chitinase and N-acetylhexosaminidase activities in submerged cultures of Verticillium fungicola increased up to 5-fold and 2.5-fold, respectively when the pH of the culture medium was raised from 5 to 8. SDS-PAGE and zymograms of the freeze-dried crude enzyme obtained from the cultures indicated four chitin degrading proteins of M(w) 24, 40, 55 and 63 kDa, whereas isoelectric focusing displayed the separation of three chitin degrading enzymes with isoelectric points of 4.7, 6.8 and 10, as well as two N-acetylhexosaminidases having isoelectric points of 3.2 and 13. Freeze-dried crude enzyme was characterized for its ability to produce chito-oligosaccharides from chitosans. Matrix-assisted laser desorption ionization time of flight mass spectrometry analyses revealed that monomers as well as hetero-oligomers with degree of polymerization 4 were initially the main products, whereas oligomers with degree of polymerization 2-11 were detected after extended reaction times.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Quitina/metabolismo , Quitinasas/biosíntesis , Verticillium/citología , Verticillium/enzimología , beta-N-Acetilhexosaminidasas/biosíntesis , Electroforesis en Gel de Poliacrilamida , Concentración de Iones de Hidrógeno , Hidrólisis , Focalización Isoeléctrica , Fracciones Subcelulares/enzimología , Factores de Tiempo
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