Compositions and methods for inhibiting the growth of fungi

Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing

Reexamination Certificate

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C424S404000, C424S405000

Reexamination Certificate

active

06280725

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to antifungal compositions and to methods for applying such compositions, in particular in the preservation of food against deterioration by fungal growth.
BACKGROUND OF THE INVENTION AND STATE OF THE ART
It is known that a variety of compounds have fungistatic properties and can therefore be used to inhibit the growth of fungi in a product. However, many fungistatic compounds are generally not accepted in food products or they have a non-natural image.
WO 97/16973 (Unilever) discloses compositions suitable for combating fungi in food and other products such as personal care products, comprising the combination of a fungal cell wall lytic enzyme and a natural microbial membrane affecting substance (MMAS), in an effective concentration. The preferred enzymes are chitinase, &bgr;-(1,3)-glucanase and &bgr;-(1,6)-glucanase. The MMAS is exemplified by nisin, amphiphilic alpha-helix forming peptides, such as histatins and the polypeptide FASLLGKALKALAFQ (=Phe-Ala-Ser-Leu-Leu-Gly-Lys-Ala-Leu-Lys-Ala-Leu-Ala-Lys-Gln) (SEQ ID No:1), and fungal inhibitors, such as carvacrol and sorbic acid. MMAS are present in herbs suitable for food preparation and also herbs suitable for cheese manufacture, especially chives, curcuma and garlic.
However, further research has shown that the fungistatic compounds described in WO 97/16973 are not always heat stable so that they become partly or completely inactivated when the products incorporating them are heated to achieve pasteurisation or even sterilisation.
WO 97/14310 describes the combination of saccharides in general with biocidal active component for combating plant pathogens.
EP 867125 describes combinations of water-soluble hemicellulose in general with a component from a range of preservatives, for preserving food and drinks.
WO 93/04588 discloses combinations of chelating agents and lanthionines (amongst which is nisin) as antimicrobial compositions. Among the chelating agents are mentioned polyols, saccharides, polyethylene glycol.
JP 63/185358 describes combinations of starch hydrolysates with one of glycine, maltose, ethanol, sorbic acid, lysozyme, edible salts of acetic acid for preservation.
Chemical Abstracts 116 (11), Mar. 16, 1992 (Kurusawa et al) describe the induction of cellulase by &bgr;-gentiobiose.
Thus, in spite of the success of the approach described above, there is still a need for fungistatic compounds that are sufficiently heat-stable in addition to being acceptable in food products and other products that come in contact with humans, such as personal care products.
SUMMARY OF THE INVENTION
In accordance with the present invention there is provided a composition suitable for inhibiting the outgrowth of fungi characterised in that it comprises a first ingredient which inhibits the biogenesis of a normal fungal cell wall and a second ingredient which is capable of perturbing the structure of the cellular membrane of said fungi, so that either the cellular integrity is essentially lost or cell division cannot take place, or both.
In one aspect of the invention, the first ingredient inhibits the anchorage of cell wall proteins into the cell wall of the fungi. This first ingredient is preferably selected from the group consisting of &bgr;-(1,6)-glucose polysaccharides, branched polysaccharides having a &bgr;-(1,6)-glucose-backbone, and mixtures thereof. Preferred compounds include &bgr;-gentiobiose and pustulan fragments.
In another aspect of the present invention, the second ingredient is preferably selected from the group consisting of natural microbial membrane affecting substance (MMAS). Preferred compounds are nisin, amphiphilic alpha-helix forming peptides such as MB-21, and fungal inhibitors present in herbs suitable for food preparation, such as carvacrol and sorbic acid.
In a further embodiment of the present invention, the first ingredient is preferably present in a concentration of 0.05 to 20 wt. %, more preferably 0.1 to 1 wt. %, calculated on the composition. The second ingredient is preferably present in a concentration of 5×10
−5
to 1×10
−2
wt. %, more preferably 1×10
−4
to 2.5×1/
−3
wt. %, calculated on the composition.
The compositions of the present invention are suitably used in food products, such as sauces, sauce-bases, dressings, ketchups, soups, soup-bases, spreads, beverages (e.g. tea-based beverages), ice cream, and in personal care products such as skin creams, lotions, ointments, as well as shampoos and other products for application to hair.
These and other aspects will be set out in more detail in the following description and claims.


REFERENCES:
patent: 5888504 (1999-03-01), Brul et al.
patent: 0 867 125 A1 (1996-09-01), None
patent: 63185358 (1988-05-01), None
patent: WO 93/04588 (1993-03-01), None
patent: WO 96/28468 (1996-09-01), None
patent: WO 97/01348 (1997-01-01), None
patent: WO 97/14310 (1997-04-01), None
patent: WO97/16973 (1997-05-01), None
Granot et al. Carbon Source Induces Growth of Stationary Phase Yeast Cells, Independent of Carbon Source Metabolism; Yeast, vol. 9, pp. 465-479, 1993.*
Kurasawa et al.Appl. Environ. Microbiol., 58(1), 106-110 (1992), inChem Abst, 116, 102396.

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