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Applied and Environmental Microbiology, March 2000, p. 1066-1076, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Prevention of Yeast Spoilage in Feed and Food by the Yeast Mycocin HMK

K. F. Lowes,1,2 C. A. Shearman,1,* J. Payne,1 D. MacKenzie,1 D. B. Archer,1 R. J. Merry,2 and M. J. Gasson1

Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA,1 and Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB,2 United Kingdom

Received 29 June 1999/Accepted 24 November 1999

The yeast Williopsis mrakii produces a mycocin or yeast killer toxin designated HMK; this toxin exhibits high thermal stability, high pH stability, and a broad spectrum of activity against other yeasts. We describe construction of a synthetic gene for mycocin HMK and heterologous expression of this toxin in Aspergillus niger. Mycocin HMK was fused to a glucoamylase protein carrier, which resulted in secretion of biologically active mycocin into the culture media. A partial purification protocol was developed, and a comparison with native W. mrakii mycocin showed that the heterologously expressed mycocin had similar physiological properties and an almost identical spectrum of biological activity against a number of yeasts isolated from silage and yoghurt. Two food and feed production systems prone to yeast spoilage were used as models to assess the ability of mycocin HMK to act as a biocontrol agent. The onset of aerobic spoilage in mature maize silage was delayed by application of A. niger mycocin HMK on opening because the toxin inhibited growth of the indigenous spoilage yeasts. This helped maintain both higher lactic acid levels and a lower pH. In yoghurt spiked with dairy spoilage yeasts, A. niger mycocin HMK was active at all of the storage temperatures tested at which yeast growth occurred, and there was no resurgence of resistant yeasts. The higher the yeast growth rate, the more effective the killing action of the mycocin. Thus, mycocin HMK has potential applications in controlling both silage spoilage and yoghurt spoilage caused by yeasts.

* Corresponding author. Mailing address: Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. Phone: 44-(1)603 255000. Fax: 44-(1)603 507723. E-mail: claire.shearman{at}bbsrc.ac.uk.

Applied and Environmental Microbiology, March 2000, p. 1066-1076, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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