Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing
Patent
1995-02-22
1998-07-21
Carlson, Karen
Drug, bio-affecting and body treating compositions
Enzyme or coenzyme containing
435183, A61K 3748, C12N 900
Patent
active
057831831
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention discloses a method for preserving beverages comprising the use of a killer toxin. Specifically an effective amount of a zymocin is used. The zymocin is particularly used in carbonated drinks. The invention also discloses the cloning of the Williopsis mrakii DNA and the DNA sequence encoding zymocin. This makes possible overexpression and large scale production of zymocin.
BACKGROUND OF THE INVENTION
Spoilage of food by yeasts is a well known problem in food industry. In some products yeasts are the main source of spoilage. Fruits belong to the natural habitat of yeasts. Therefore, it is not surprising that fruit derived products and fruit containing products are susceptible to contamination by yeasts. Examples are fruit pulp, fruit juice, fruit containing softdrinks and carbonated beverages, which sometimes contain high percentages of fruit juice. Spoilage of those products is even a major problem in industry. Softdrinks and carbonated beverages are to be considered as a very selective environment for the growth of yeasts. This environment has peculiar physical characteristics specifically low pH, low oxygen concentration and high sugar concentration. Furthermore storage is at low temperature. Yeasts have the ability to ferment the sugars introduced as such or those introduced with the fruits. Outgrowth of yeasts is responsible for off-flavours, there is loss of texture quality and gas is produced. The production of gas can cause swelling or even blowing up of the container. In the case of carbonated beverages bottled in glass this phenomenon has caused severe injuries. Sellar. P. W. and P. B. Johnston (BMJ (1991) 303 176-177)) describe how the seriousness of ocular injuries caused by exploding bottles has been underestimated. T. Willhoft (New Scientist (1986) 21 Aug., 28-30) also reports the popping bottle problem. The problem is due to carbon dioxide and although not specifically reported it is known that this gas is also produced by yeasts.
Yeast genera most frequently isolated from fruit and fruit products are Saccharomyces, Kluvveromyces, Zygosaccharomyces, Debaryomyces, Hansenula, Candida, Pichia and Torulopsis. The yeasts are introduced into the products with the ingredients such as the fruits. The yeasts also may grow on the surfaces of the production equipment.
In spite of good hygienic production methods and the use of preservatives like sorbate and benzoate spoilage of fruit products is still very common. Chemical preservatives like sorbic acid and benzoic acid pose their own problems. The concentration of these chemicals needed to prevent the outgrowth of some yeast species commonly detected in fruit derived products is very high. The concentration of benzoate and/or sorbate permitted in those products is too low to prevent outgrowth of these yeasts.
Killer toxins are inhibitory molecules produced by a wide range of yeasts. The inhibitory molecules are polypeptides or polypeptide containing molecules which kill sensitive yeast cells. Killer toxins were first observed in certain strains of the genus Saccharomyces (Makower M. et al. (1963) Proc. 11th Int. Congr. Genet. I, 1202). Today many different killer toxin-producing yeast species are known (Young T. W. (1987) in The Yeasts, Rose A. H. et al. Eds. Acad. Press, London. 2: 131-164). The majority of the killer toxins are not very stable and therefore not useful as a preservative. However, various Hansenula species make an exception to this rule and produce very heat- and pH-stable killer toxins (Nomoto, H. et al. (1984) Agric. Biol. Chem. 48 : 807-809).
For commercial use as an anti-yeast preservative it is preferred to employ a stable killer toxin with a broad spectrum anti-yeast activity.
An example of such a killer toxin is the killer toxin produced and secreted by Hansenula mrakii IFO 0895. The killer toxin of Hansenula mrakii IFO 0895 is very stable against heat and in a pH range of 4-11. A broad spectrum activity was demonstrated (Ashida S (1983) Agric. Biol. Chem. 47: 2953-2955). Further characte
REFERENCES:
patent: 4695468 (1987-09-01), Boston
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La Roceo etal 1985 Food Technology 39(7):49-52.
Martin etal 1984 Microbios Letters 26:115-119.
Sellar et al., "Ocular Injuries Due to Exploding Bottles of Carbonated Drinks", BMJ (1991) 202:176-177.
Willhoft, "Victims of the Pop Bottle", New Scientist, 21 Aug. 1986, pp. 28-30.
Makower et al., "The Inheritance of a Killer Character in Yeast (Saccharomyces cereviseae)" (1963) Proc. 11th Int. Congr. Genet. I. 1202.
Young, (1987) in The Yeasts, Rose A.H. et al. Eds. Acad. Press, London, 2:131-164.
Nomoto et al., "Distribution of Killer Yeasts in the Genus Hansenula", Agric, Biol. Chem. 48:807-809 (1984).
Ashida et al., "New Killer Toxin of Hansenula mrakii", Agric. Bio. Chem. 47:2953-2955 (1983).
Yamamoto et al., "Application of Monoclonal Antibodies to the Isolation and Characterization of a Killer Toxin Secreted by Hansenula mrakii" (1986) FEBS 195:253-257.
Ohta et al., "Production, Purification and Characterization of HYI, an Anti-yeast Substance, Produced by Hansenula saturnus", Agric. Biol. Chem. 48:903-908 (1984).
Langeveld Pieter Cornelis
Solingen Pieter Van
Stark Jacobus
Vollebregt Adrianus Wilhelmus Hermanus
Carlson Karen
Gist-Brocades B.V.
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