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Applied and Environmental Microbiology, May 2000, p. 1809-1813, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

(1right-arrow 6)-beta -D-Glucan as Cell Wall Receptor for Pichia membranifaciens Killer Toxin

A. Santos,1 D. Marquina,1,* J. A. Leal,2 and J. M. Peinado1

Department of Microbiology, Biology Faculty, Complutense University of Madrid, Madrid 28040,1 and Centro de Investigaciones Biológicas (CSIC), Madrid 28006,2 Spain

Received 17 September 1999/Accepted 15 February 2000

The killer toxin from Pichia membranifaciens CYC 1106, a yeast isolated from fermenting olive brines, binds primarily to the (1right-arrow6)-beta -D-glucan of the cell wall of a sensitive yeast (Candida boidinii IGC 3430). The (1right-arrow6)-beta -D-glucan was purified from cell walls of C. boidinii by alkali and hot-acetic acid extraction, a procedure which solubilizes glucans. The major fraction of receptor activity remained with the alkali-insoluble (1right-arrow6)-beta - and (1right-arrow3)-beta -D-glucans. The chemical (gas-liquid chromatography) and structural (periodate oxidation, infrared spectroscopy, and 1H nuclear magnetic resonance) analyses of the fractions obtained showed that (1right-arrow6)-beta -D-glucan was a receptor. Adsorption of most of the killer toxin to the (1right-arrow6)-beta -D-glucan was complete within 2 min. Killer toxin adsorption to the linear (1right-arrow6)-beta -D-glucan, pustulan, and a glucan from Penicillium allahabadense was observed. Other polysaccharides with different linkages failed to bind the killer toxin. The specificity of the killer toxin for its primary receptor provides an effective means to purify the killer toxin, which may have industrial applications for fermentations in which salt is present as an adjunct, such as olive brines. This toxin shows its maximum killer activity in the presence of NaCl. This report is the first to identify the (1right-arrow6)-beta -D-glucan as a receptor for this novel toxin.

* Corresponding author. Mailing address: Department of Microbiology, Biology Faculty, Complutense University of Madrid s/n, 28040 Madrid, Spain. Phone and Fax: (34-91)394 49 64. E-mail: dommarq{at}eucmax.sim.ucm.es.

Applied and Environmental Microbiology, May 2000, p. 1809-1813, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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