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Applied and Environmental Microbiology, December 2000, p. 5348-5352, Vol. 66, No. 12
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Isolation, Purification, and Characterization of a Killer Protein from Schwanniomyces occidentalis

Wen-Bao Chen,1 Yuh-Fehng Han,1 Shung-Chang Jong,2 and Shenq-Chyi Chang1,*

Department of Biochemistry, National Yang-Ming University, Taipei 112, Taiwan, Republic of China,1 and Mycology Department, American Type Culture Collection, Manassas, Virginia 201102

Received 9 May 2000/Accepted 4 October 2000

The yeast Schwanniomyces occidentalis produces a killer toxin lethal to sensitive strains of Saccharomyces cerevisiae. Killer activity is lost after pepsin and papain treatment, suggesting that the toxin is a protein. We purified the killer protein and found that it was composed of two subunits with molecular masses of approximately 7.4 and 4.9 kDa, respectively, but was not detectable with periodic acid-Schiff staining. A BLAST search revealed that residues 3 to 14 of the 4.9-kDa subunit had 75% identity and 83% similarity with killer toxin K2 from S. cerevisiae at positions 271 to 283. Maximum killer activity was between pH 4.2 and 4.8. The protein was stable between pH 2.0 and 5.0 and inactivated at temperatures above 40°C. The killer protein was chromosomally encoded. Mannan, but not beta -glucan or laminarin, prevented sensitive yeast cells from being killed by the killer protein, suggesting that mannan may bind to the killer protein. Identification and characterization of a killer strain of S. occidentalis may help reduce the risk of contamination by undesirable yeast strains during commercial fermentations.

* Corresponding author. Mailing address: Department of Biochemistry, National Yang-Ming University, Taipei 112, Taiwan, Republic of China. Phone: 886-2-28267120. Fax: 886-2-28264843.

Applied and Environmental Microbiology, December 2000, p. 5348-5352, Vol. 66, No. 12
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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