1887

Abstract

Summary: The presence of non-fibrillar α-chitin in celluiosic fungi (class Oomycetes) poses intriguing questions as to its role, subcellular localization and evolutionary significance. Previous studies reported on the similarity of chitin synthase from with that of other fungi. The present work describes important dissimilarities. There was no evidence that the chitin synthase of was present in small low-density vesicles (chitosomes). Chitin synthase sedimented with membranous components of high specific gravity (sp. gr. 1.177) that could be partially but distinctly separated from membranes harbouring most of the 1,3–glucan synthase in the cell csp. gr. 1.158). In contrast to other fungi, the chitin synthase from was greatly stimulated by digitonin: both membrane-bound and dissociated chitin synthase showed little activity in the absence of digitonin. As in other fungi, the chitin synthase from was solubilized by digitonin and remained zymogenic after dissociation. However, unlike the enzyme from other fungi, the solubilized chitin synthase of had a lower sedimentation coefficient, was not stimulated by phospholipids and was not inhibited by high concentrations of digitonin. Unlike the enzyme from the solubilized chitin synthase from did not bind to a cation exchanger. The enzyme was partially purified by a four-step scheme that included sucrose density-gradient centrifugation, a single passage through a strong anion exchanger and two consecutive passages through a weak anion exchanger. The final preparation contained five to seven polypeptide bands that cochromatographed with the chitin synthase activity, some of which may be part of a presumed chitin synthase macromolecular complex.

Funding
This study was supported by the:
  • National Institutes of Health (Award GM-48257)
  • National Science Foundation (Award IBN-9204541)
  • Centre National de la Recherche Scientifique (Award UMR-5534)
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/content/journal/micro/10.1099/00221287-143-7-2473
1997-07-01
2021-10-24
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