Antibacterial compounds

Abstract

The present invention relates to the field of antimicrobial active substances. In addition to the novel uses of the unique antimicrobial mixtures according to the invention, the invention also relates to preparations, in particular nutritional or recreational or pharmaceutical preparations, containing such mixtures, as well as methods for preparing such preparations and methods for the antimicrobial treatment of nutritional or recreational or pharmaceutical preparations.

Description

[Technical field]

[0001] The present invention relates to the field of antimicrobial active substances. In addition to the novel use of the unique antimicrobial mixture according to the invention, the present invention also relates to preparations containing such mixtures, in particular preparations used for nutrition or pleasure or pharmaceutical preparations, as well as methods for preparing such preparations and methods for the antimicrobial treatment of preparations used for nutrition or pleasure or pharmaceutical preparations. Further aspects of the present invention are derived from the following specification and in particular from the appended claims. [Background technology]

[0002] Foods, especially those with ingredients of animal origin (mayonnaise, minced meat and sausage products), as well as sugar-containing foods (sweetened drinks, syrups, ketchup and dressings), are prone to microbial spoilage due to their nutrient richness and high water activity coefficient (aw value).

[0003] To stabilize such products, typically the concept of hurdles is applied (Non-Patent Document 1): raw materials with low initial germination numbers are selected, if possible, aw values, redox potentials and pH values ​​are reduced when formulating the product, and special attention is paid to food hygiene and GMP (Good Manufacturing Practice) when processing the product. As a final hurdle, preservation methods are applied, such as pasteurization or the use of preservatives. However, traditional preservation methods are increasingly viewed as questionable by consumers. For example, pasteurization can reduce the amount of nutrients by creating a cooked feeling and impair the taste. Preservatives such as sorbic acid and benzoic acid are obtained by synthetic methods and must be declared as additives. Natural preservatives such as acidic acids, poppy seeds, essential oils and their extracts can only be applied within certain limits, since they strongly alter the product sensorially.

[0004] The growth of bacteria is strongly inhibited by pH values ​​below 4.5, except for acidophilic bacteria such as Lactobacillus sp. or Acetobacter sp., but yeasts and fungi can also grow at pH values ​​below 3, leading to spoilage. Special yeasts such as Zygosaccharocyces baili, Zygosaccharomyces lentus and Saccharomyces cerevisiae also show resistance to preservatives such as sorbic acid and benzoic acid (Non-Patent Document 2, Non-Patent Document 3). Food spoilage caused by yeasts and fungi is the most difficult to control, but what is known is mainly the antimicrobial effect of plant extracts (Non-Patent Document 4). For example, herbs, spices and essential oils and extracts obtained therefrom have a confirmed effect against bacteria (5), but due to their own aroma they unacceptably alter the taste of many products. Only a few plant extracts are known to have antibacterial effects against yeasts and fungi, and their use in foods is often limited by their own aroma, strong coloring properties and low solubility. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Kraemer, J., Alexander, P., (2017), Food Microbiology, Stuttgart, Eugen Ulmer, 7th revised edition. [Non-Patent Document 2] Steels, H., James, SA, Roberts, IN and Stratford, M. (1999), "Zygosaccharomyces lentus: a significant new osmophilic, preservative-resistant spoilage yeast, capable of growth at low temperature", Journal of Applied Microbiology, Vol. 87, No. 4, pp. 520-527. [Non-Patent Document 3] Juvonen, R., Virkajaervi, V., Priha, O. and Laitila, A., (2011), "Microbiological spoilage and safety risks in non-beer beverages", Newsletter of the Finnish National Institute of Science (VTT Tiedotteita) - Research notes No. 2599. [Non-Patent Document 4] Friedman, M. (2015), "Antibiotic-resistant bacteria: prevalence in food and inactivation by food-compatible compounds and plant extracts." J Agric Food Chem, Vol. 63, No. 15, pp. 3805-3822. [Non-Patent Document 5] Weber, H. (2010), "Food Microbiology - Fundamentals", Hamburg, Behrs Verlag. Summary of the Invention [Problem to be solved by the invention]

[0006] It was therefore the aim and primary objective of the present invention to develop an alternative formulation for increasing the shelf life of fresh food products without undue technical effort, preferably by the addition of natural plant-based ingredients, and thereby preferably not adversely affecting the taste, or ideally even completing or improving the taste, respectively.

[0007] Furthermore, the formulations to be provided must be toxicologically harmless, highly compatible, stable (especially in typical formulations), and / or inexpensive to manufacture.

[0008] It would be further desirable to use as little antimicrobial active material as possible in each formulation to achieve a particular antimicrobial effect.

[0009] The search for suitable substances having sufficient amounts of one or more of the above-mentioned properties is further complicated for those skilled in the art by the absence of a clear dependency between the chemical structure of a substance and its biological activity against specific microorganisms (germs) as well as its stability. Moreover, there is no predictable relationship between the antimicrobial effectiveness, toxicological harmlessness and stability of a substance.

[0010] It was therefore a preferred object of the present invention to provide a formulation which meets some or preferably all of the above criteria, or which has a desirable combination of the above properties. [Means for solving the problem]

[0011] This first objective is to (A) Plants of the following families: Asparagaceae, Cannabaceae, Ericaceae, Lythraceae, Krameriaceae, Rosaceae, and Vitaceae. Preferably, the following plants are used: hops (Humulus lupulus), cultivated strawberry (Fragaria x anapassa), Tribulus terrestris (Krameria lappacea), pomegranate (Punica granatum), lowbush blueberry (Vaccinium angustifolium), mountain cranesbill (Vaccinium macrocarpon), Vaccinium myrtillus, bilberry (Vaccinium vitis-idaea), and Vitis vinifera. extracts and fractions of those extracts, one or more plant extracts having antibacterial effect and / or one or more fractions of said extracts having antibacterial effect; (B) abrusoside and balancine (preferably as described in WO 2012 164,062), malic acid, benzaldehyde, gamma-decalactone, delta-decalactone, 3,7'-dihydroxy-4'-methoxyflavan isomers (preferably as described in EP 2,253,226), eriodictyol, ethyl 2-methylbutyrate, ethyl butyrate, ethyl caproate, geranial, hesperetin (preferably as described in WO 2007 / 014879, EP 2,368,442(B1) or EP 1,909,599(B1)), or sweet tea (Rubus suavissimus) suavissimus) extracts (preferably those described in U.S. Provisional Patent Application No. 61 / 333,435 and patent applications thereunder), hesperetin dihydrochalcone, hesperidin dihydrochalcone, hydroxybenzoic acid amides (preferably those described in WO 2006 / 024587), such as 2,4-dihydroxybenzoic acid vanillylamide, 4-hydroxydihydrochalcone (preferably those described in U.S. Patent Application Publication No. 2008 / 0227867(A1) and WO 2008 / 0227867(A1), 2007 / 107596), e.g. phloretin and davidigenin, p-hydroxybenzaldehyde, p-hydroxybenzoic acid, homoeriodictyol, limonene, lactic acid, menthofloractone, mogrosides, naringenin, naringin dihydrochalcone, neoisoflavonoids (preferably those described in EP Patent No. 2,570,036 (B1)), neohesperidin dihydrochalcone, neral, δ-octalactone, pellitorinin (preferably those described in EP Patent No. 2,570,036 (B1)), 008 530(A1) or the fragrance compositions described therein and fragrance compositions derived therefrom), piperonal, phlorididine, phyllodulcin isomers (preferably those described in EP Patent No. 2,298,084(B1), rubusoside (preferably those described in EP Patent No. 2,386,211), mixtures of rubusoside isomers and homologues, 1-(2,4-dihydroxy-phenyl)-3-(3-hydroxy-4-methoxy-phenyl)-propan-1-one (preferably those described in EP Patent No. 2,353,403(B1)), stevioside and rebaudioside (preferably those described in WO 2015 062,998), trilobatin, vanillin, vanillic acid (preferably those described in European Patent Application having serial number 2,517,574), vanillyl lignans, tartaric acid and cinnamon aldehyde, one or more flavouring agents which have an antimicrobial effect and / or which enhance the antimicrobial efficacy of component (A) and / or which reduce, mask or modify the unpleasant taste impression of component (A) and / or which enhance or modify the pleasant taste impression of component (A); The problem is solved by the use of a mixture comprising or consisting of as an antibacterial mixture, preferably a synergistic antibacterial mixture.

[0012] According to an embodiment of the present invention, for one or more plant extracts and / or one or more fractions of these extracts having an antibacterial effect according to (A), preferably the roots and / or fruiting organs of the plants or parts thereof, preferably the endocarp, mesocarp, exocarp and / or flower head parts, are used.

[0013] The substances or parts of those substances according to (B) are, according to an embodiment of the invention, preferably selected from the following plants: Artemisia xanthochroa, tarragon, Bauhinia manca, Chenopodium album, Chrysothamnus ssp., citrus spp., in particular insbesondere, orange, Citrus sinensis, grapefruit, bergamot, Dracaena cinnabaria (cinnabari), Herba santa, Hydrangea (Hydrangeae) dulcis, Mexican sweet herb, Lippia dulcis, Lycoris aurea, Malus domestica, Mariscus psilostachys, Momordica grosvenorii, Piper spp., Stevia ssp., Vanilla spp., Zanthoxylum rubescens extracts (or fractions of said extracts).

[0014] According to the present invention, component (B) is - improve the antimicrobial effect of component (A) (synergistic antimicrobial effect, i.e. the antimicrobial effect exceeds the sum of the antimicrobial effects of the substances with antimicrobial effect in the mixture), which advantageously makes it possible to use lower (lower) amounts of the substances with antimicrobial effect in the mixture, and / or - reducing, masking or modifying the unpleasant taste impression of component (A), and / or -Enhancing or modifying the pleasant taste impression of component (A) Such antibacterial mixtures are preferred.

[0015] Thus, according to the present invention, it is preferred that component (B) is present in the mixtures used according to the present invention in a total amount sufficient to enhance the antimicrobial effect of component (A) and / or to reduce, mask or modify the unpleasant taste impression of component (A) and / or to increase or modify the pleasant taste impression of component (A).

[0016] In this specification, the taste impression to be reduced, masked or modified is preferably selected from the group consisting of astringent, bitter, medicinal, metallic, sour.

[0017] The taste impression to be enhanced or modified is preferably selected from the group consisting of sweet, juicy and full-bodied.

[0018] The mixtures to be used according to the invention are highly suitable as antimicrobial mixtures of active substances for increasing the shelf life of fresh compositions.

[0019] Advantageously, the mixtures to be used according to the invention also make it possible to fulfil the initially mentioned criterion.

[0020] Although experts have already considered the antibacterial properties of the components described herein in part, there has been no specific indication until now that the mixtures to be used according to the invention have the effects and advantages described herein. The significantly improved antibacterial effect, especially against the bacteria described herein, is particularly surprising.

[0021] The present invention relates to a method for the preparation of a mixture according to the invention, which is particularly suitable for the preparation of a fungus or yeast, in particular Dekkera bruxellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomyces ludwigii, Zygosaccharocyces bailii and Pichia sp., Aureobasidium sp., Alternaria alternata, Aspergillus brasiliensis, Aspergillus fu-migatus (fumigatus), Cladosporium sp., Fusarium sp., Rhodotorula sp., Sporidiobolus sp., Sporobolomyces sp., Paecilomyces sp. and Penicillium sp. The present invention is based on the surprising realization that each of the compounds exhibits a particularly advantageous antibacterial effect against one or more bacteria selected from the group consisting of:

[0022] Therefore, the use according to the invention against fungi or yeasts, preferably against one or more or all bacteria selected from the group consisting of S. cerevisiae, Z. bailii, C. albicans, A. pullulans and A. braziliensis, respectively, is particularly preferred - and advantageous.

[0023] According to the invention, it is preferred if the extract of component (A) is selected from the group consisting of berry extracts, preferably red berry extracts, pomegranate extracts and herbal extracts.

[0024] Extracts of component (A) can be prepared from whole plants, plant parts, biotechnologically altered or unaltered plant material, using methods known to those skilled in the art, preferably solid-liquid extraction methods. Furthermore, if necessary, enrichment and concentration of selected substances may also be applied after the extraction has been carried out.

[0025] For example, extraction of dried plant parts may apply pure or mixtures of solvents such as acetone, butane, butan-1-ol, butan-2-ol, cyclohexane, nitrous oxide, diethyl ether, ethyl acetate, ethanol, ethyl methyl ketone, hexane, carbon dioxide, methanol, methyl acetate, vegetable oils, propane, propan-1-ol, propan-2-ol and / or water.

[0026] To improve the yield, additives such as surface active substances, hygroscopic substances and / or salts may be added to the extraction solvent.

[0027] Moist plant parts may be extracted using microwave radiation, ultrasound and pulsed electric fields.

[0028] Fractionation of the extract (production of the fractions described herein) is preferably carried out by separation of undesired minor components and / or concentration of selected substances, e.g. by adsorptive distillation, chromatographic separation, fermentation, filtration, liquid-liquid extraction, membrane filtration and / or centrifugal sedimentation, preferably by chromatographic separation, liquid-liquid extraction and / or membrane filtration.

[0029] Following fractionation of the extract, the components of each fraction may be determined, where appropriate, using known methods. Determination of the compounds using mass spectrometry, particularly LC-MS, is a preferred method.

[0030] According to the invention, it is particularly preferred that component (A) comprises at least one, two, three or more compounds selected from the group consisting of catechins, chlorogenic acid, p-coumaric acid, epicatechin, ellagic acid, gallic acid, caffeic acid, sinapic acid and cinnamic acid.

[0031] Depending on the fractionation method, in particular the ratio of the concentrations of the substances catechin, chlorogenic acid, p-coumaric acid, epicatechin, ellagic acid, gallic acid, caffeic acid, sinapic acid and / or cinnamic acid relative to the other components may be altered by fractionation in a targeted manner, whereby the alterations are preferably applied so as to improve the antimicrobial effect.

[0032] In yet another preferred embodiment, component (B) contains or consists of at least one, two, three or more flavorings selected from the group consisting of eriodictyol, hesperetin, homoeriodictyol, naringenin, pellitoriin and vanillic acid.

[0033] For the mixtures to be used according to the invention, preferably the ratio of the total amount of component (A) to the total amount of component (B) applies in the range of 1000:1 to 1:10, preferably 100:1 to 1:10, preferably 100:1 to 1:1, preferably 10:1 to 1:1. With respect to this feature, preferably the amount of a candidate component which can be allocated to both component (A) and to component (B) applies to be allocated to the total amount of component (A).

[0034] Advantageously, the mixtures described herein are particularly suitable for preparations used for nutritional or palatable purposes or for pharmaceutical preparations.

[0035] The present invention therefore also relates to preparations used for nutritional or enrichment purposes or pharmaceutical preparations comprising the mixtures described herein, wherein the mixtures are present in an amount sufficient to achieve an antimicrobial effect, preferably a synergistic antimicrobial effect.

[0036] What has been said above about the mixtures to be preferably used according to the invention applies accordingly to the preferred embodiments of the mixtures involved.

[0037] Thus, the mixture is Dekkera brucellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomycodes ludwigii, Zygosaccharomyces bailii and Pichia spp. Aureobasidium spp., Alternaria alternata, Aspergillus braziliensis, Aspergillus fumigatus, Cladosporium spp., Fusarium spp., Rhodotorula spp., Sporidiobolus spp., Sporobolomyces spp., Paecilomyces spp. and Penicillium spp., It is preferred that the composition is contained in an amount sufficient to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more bacteria selected from the group consisting of:

[0038] It is particularly preferred if the mixture is contained in an amount sufficient to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailiii, C. albicans, A. pullulans and A. braziliensis.

[0039] The use of the mixture used according to the invention in the preparation according to the invention is not limited thereby.Therefore, it is possible to envisage a number of different preparations used for nutrition or taste or a number of different pharmaceutical preparations.Particularly preferably, the preparation according to the invention is food, especially those with ingredients of animal origin (such as mayonnaise, minced meat and sausage products) and sugar-containing food (sweetened drinks, syrups, ketchups and dressings).

[0040] According to a preferred embodiment of the invention, the mixture to be used according to the invention is therefore applied in a food product, preferably a food product having components of animal origin, such as mayonnaise, minced meat and sausage products, and sugar-containing food sweetened beverages, such as syrups, ketchups, cream dressings and dressings.

[0041] The mixtures to be used according to the invention may, for example, be contained in a total amount of from 3 to 0.001, preferably from 0.5 to 0.01, particularly preferably from 0.1 to 0.01% by weight, relative to the total weight of the preparation.

[0042] For these purposes, the mixture to be used according to the invention may be present in such preparations in the form of liquid components or powders or spray-dried products. Thus, for example, components (A) and (B) are mixed, dissolved and added in liquid or spray-dried form. Components (A) and (B) may also be used separately, in different forms. The preparation according to the invention also preferably further comprises components typical of such preparations. For example, a solvent or carrier may be present.

[0043] Here, the solvent may be, for example, water, ethanol, 1,2-propanediol, triacetin, diacetin, triethyl citrate and / or glycerin.

[0044] Advantageous carriers are, for example, silicon dioxide (silica acid, silica gel), carbohydrates and / or carbohydrate polymers (polysaccharides), cyclodextrins, starches, degraded starches (starch hydrolysates), chemically or physically modified starches, modified celluloses, gum arabic, gum ghatti, gum tragacanth, gum karaya, carrageenan, guar gum, locust bean gum, alginates, pectins, inulin or xanthan gum. Preferred starch hydrolysates are maltodextrins and dextrins, with maltodextrins having a DE value ranging from 5 to 20 being particularly preferred. It is not important which plant originally provided the starch for producing the starch hydrolysate. Corn-based starches as well as tapioca, rice, wheat or potato starches are suitable and are easily accessible. The carrier may also serve as a flow additive, such as, for example, silicon dioxide.

[0045] One aspect of the present invention is (i) providing a mixture as described herein; (ii) providing one or more further ingredients suitable for consumption; (iii) mixing the components provided in steps (i) and (ii); The present invention also relates to a method for producing the preparation according to the invention, comprising or consisting of the steps:

[0046] The above statements regarding the mixtures to be preferably used according to the invention also apply to the preferred embodiments of the mixtures contained therein. The same applies, where appropriate, to the preferred embodiments of the preparations to be produced.

[0047] (i) providing a mixture as described herein; (ii) providing the preparation to be processed, or one or more further components of the preparation to be processed, which components are suitable for consumption; (iii) adding said mixture to a prepared preparation or ingredients suitable for consumption of the preparation. In order to achieve an antimicrobial effect, preferably a synergistic antimicrobial effect, Preferably, Dekkera brucellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomycodes ludwigii, Zygosaccharomyces bailii and Pichia spp. Aureobasidium, Alternaria alternata, Aspergillus braziliensis, Aspergillus fumigatus, Cladosporium, Fusarium, Rhodotorula, Sporidiobolus, Sporobolomyces, Paecilomyces and Penicillium, In order to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more bacteria selected from the group consisting of: More preferably, to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailiii, C. albicans, A. pullulans and A. brasiliensis, adding each in sufficient quantity; Particularly preferred is also a method according to the invention for the antimicrobial treatment of a preparation used for food or leisure or a pharmaceutical preparation, preferably as described above, which comprises or consists of the steps:

[0048] The above statements regarding the mixtures to be preferably used according to the invention also apply to the preferred embodiments of the mixtures contained therein. The same applies accordingly to the preferred embodiments of the preparations to be treated.

[0049] In the following the invention is further illustrated by selected examples, without thereby limiting the subject matter of the invention. Unless otherwise indicated, all % indications relate to weight (wt-%). EXAMPLES

[0050] 1. Pomegranate Extract (GRA): A pomegranate peel extract is produced by halving the pomegranate fruit, squeezing it, and then homogenizing the remaining peel mash with hot water at a temperature of 50° C. The mash is then squeezed. The resulting extract (filtrate) is then concentrated by distillation.

[0051] The resulting concentrated extract contains, inter alia, the following components:

[0052] [Table 1] 1 Determination of surface percentage by UPLC-NQAD

[0053] For the subsequent preparation of single fractions of concentrated extract, for example membrane filtration may be applied.

[0054] For the preparation of the membrane filtrate from the pomegranate peel extract, 26 cm 2 A polyethylene glycol-based flat membrane with a membrane surface of 1000 and a cut-off of 2500 Da was rinsed twice for 20 min. A 1% extract solution (200 g) was prepared in a feed vessel, heated to 30° C. and pressurized at 30 bar. After a permeation volume of 138.23 g, it was diluted with 100 g of water and filtered again. The membrane filtrate was freeze-dried to obtain a permeate with a yield of 16.8% and a retentate with a yield of 65.8%.

[0055] The components of the obtained fractions were subsequently examined by mass spectrometry. The obtained fractions contain, among others, the following non-volatile components:

[0056] [Table 2]

[0057] 2. Red Berry Extract – Mixture of Crassula gracilis, Lowbush Blueberry and Cultivated Strawberry (ROBE): The extract is produced by mixing the fruits of the giant cranesbill, cultivated strawberry and lowbush blueberry, followed by squeezing to obtain a filtrate, which is then concentrated by distillation.

[0058] The resulting concentrated extract contains, inter alia, the following components:

[0059] [Table 3] 1 Determination of surface percentage by UPLC-NQAD

[0060] 3. European Grape Extract (Grape Seed Extract): First, Grape seeds The extract is produced by grinding the sorghum. These are then extracted with water at 50°C and filtered to obtain the extract. The resulting extract is then concentrated by distillation.

[0061] The resulting concentrated extract contains, inter alia, the following components:

[0062] [Table 4] 1 Determination of surface percentage by UPLC-NQAD

[0063] For the subsequent preparation of single fractions of concentrated extract, for example membrane filtration may be applied.

[0064] Grape seeds For the preparation of the membrane filtrate from the extract, 26 cm 2A polyethylene glycol-based flat membrane with a membrane surface of 1.0 mm and a cut-off of 2500 Da was rinsed twice at 30 bar for 20 min. A 1% extract solution (200 g) was prepared in a feed vessel, heated to 30° C. and pressurized at 30 bar. After a permeation volume of 138.23 g, it was diluted with 100 g of water and filtered again. The membrane filtrate was freeze-dried to give a permeate with a yield of 7.6% and a retentate with a yield of 58.8%.

[0065] Alternatively, or in addition, polar and non-polar fractions may be obtained, for example, by chromatographic methods.

[0066] For preparation of polar and monopolar fractions, 100 mg in 1 mL deionized water Grape seeds A solution of the extract was prepared and loaded onto a preparative scale HPLC column (PRP-1 column 250 x 21.5 mm; 10 μm particle size) previously conditioned with water / ethanol (90 / 10) at a flow rate of 10 ml / min. Deionized water and ethanol (99.5%) were used as solvents at bath temperature (isothermal at 60°C). Initially, elution was performed with water / ethanol (90 / 10) for 25 min. Then, the percentage of ethanol was increased by 10 percent to 20% in 10 min and kept at a constant level for 10 min. Subsequently, the percentage of ethanol was increased to 100% in 5 min and held for 5 min. Polar fractions were collected from 10 to 36 min run time and monopolar fractions were collected from 37 to 55 min run time. The yield of the polar fraction was 48% and the yield of the monopolar fraction was 28%.

[0067] The components of the obtained fractions were subsequently examined by mass spectrometry. The obtained fractions contain, among others, the following non-volatile components:

[0068] [Table 5] TIFF0007602789000006.tif227170

[0069] 4. Testing the antibacterial effect of different extracts: Exemplary extracts such as those described in Examples 1 to 3 were used for testing the antibacterial effect of the various extracts alone, i.e., without combination with component (B) according to the invention. For this purpose, the extracts were dissolved at 500 mg / kg in citrate buffer at pH 3. 5.63×10 4 CFU / mL of S. cerivisiae, 1.89 × 10 4 Z. Baillieu and 3.75 x 10 4 The samples were inoculated with C. albicans.

[0070] [Table 6]

[0071] For S. cerevisiae, a reduced number of germinations was observed after 24 g incubation time. After 7 days, growth ceased.

[0072] For Z. bailii, a reduction in germination was observed after 7 days, and no new colony formation occurred after 14 days.

[0073] C. albiculus was resistant to the extract for 21 days.

[0074] A. pullulans could be reduced in 28 days. For GRA, a 100% reduction could be observed after 28 days.

[0075] For A. braziliensis, a reduced number of germinations could be detected after 28 days.

[0076] As shown in the results below (see Example 7), the antibacterial effect of the extract can be enhanced by combination with component (B) described herein.

[0077] 5. Further studies on the antibacterial effects of selected extracts and fractions of those extracts: Exemplary extracts or fractions of these extracts such as those described in Examples 1 to 4 were used for testing the antibacterial effect of the various extracts alone, i.e., without combining with component (B) or fractions thereof according to the present invention. The extracts or fractions of these extracts were dissolved in citrate buffer at pH 5 in the dosages shown in the table below, respectively. 3.8×10 4 Samples were inoculated with CFU / mL of S. cerevisiae.

[0078] [Table 7]

[0079] The negative control did not result in a significant reduction in the number of germinations.

[0080] However, the positive control with 375 mg / kg potassium sorbate also did not show a significant reduction in the number of germinations. The underlying reason is that, on the one hand, sorbic acid is present in its effective undissociated form to a smaller extent compared to pH 3 in the additive example Table 2, and the optimal growth of S. cerevisiae between pH 4 and 6 occurs at pH 5.

[0081] In the case of pomegranate extract (and also in the case of Vitis vulgare extract) no significant reduction in the number of germinations of S. cerevisiae could be observed. It was therefore all the more surprising to observe an inhibition of antimicrobial growth, i.e. a log2 reduction in the number of germinations, by fractionation of the extracts: the number of germinations was significantly reduced by the addition of 200 mg / kg of the non-polar fraction of Vitis vulgare extract and by the addition of 200 mg / kg of pomegranate mebrane >2500 Da filtrate.

[0082] As shown by the results below (see Example 7), the antibacterial effect of the extract can be enhanced by combination with component (B) described herein.

[0083] 6. Interaction of extracts with flavorings:

[0084] [Table 8] JPEG0007602789000010.jpg160119

[0085] The examples show that the sensory impression of the product containing the extract described above is improved by the addition of a unique flavoring agent while maintaining the product's specific aroma and taste perception. It has been found that when combined with a flavoring agent, the undesirable side effects of the plant extract, such as bitter, astringent, or metallic, can be transferred to sensory positively perceived attributes, such as juicier, fuller, sweeter, and more complex. The same applies to the combination of the fractions of the extract as described herein with the flavoring agents described herein, where appropriate. In order to optimize the desired effect, the skilled person may test and use different dosages according to this matrix.

[0086] For further testing, non-carbonated water was used consisting of 60 mg / kg acesulfame-K, 60 mg / kg sucralose, 0.15 g / 100 g citric acid, 200 mg / kg GRA (Example 1) and 0.1 g / 100 g cherry flavor. Flavorings and flavoring mixtures with the effects described below were added and compared to the base sensory profile without the additions.

[0087] [Table 9]

[0088] In a further range of tests, the antibacterial effect of the combination of fragrance and extract was investigated. The following table shows the results of a bacterial challenge test similar to the experiments of Examples 5 and 6, except that the mixture was made with pure water at pH 7. For example: Grape seedsThe extract and the flavoring vanillic acid have different effects on yeast. However, when combined, an effect on fungi and an optimal effect on yeast were also observed. Pomegranate and red berry extracts have a lower effect at pH 7 compared to pH 3 (see Example 5), and pomegranate extract eliminates S. cerevisiae in less than 14 days at 1000 mg / kg. However, when combined with limonene-containing lemon oil, this effect can be increased for higher pH.

[0089] [Table 10] 1) The amount of limonene in lemon oil ranges from 70% to 1%.

[0090] 7. Example of use: The mixtures or combinations to be used according to the invention can each be used in several preparations, see for example the following possible applications (instead of the extracts mentioned by way of example, other extracts described herein or fractions of these extracts can also be used):

[0091] [Table 11]

[0092] 7.1 Cherry flavoured beverage (sugar free):

[0093] [Table 12] 1) The extract contains 8.3 g / kg ellagic acid and 4.7 g / kg gallic acid.

[0094] 7.2 Cherry flavoured beverages (contains sugar):

[0095] [Table 13] 1)The extract contains 8.3 g / kg ellagic acid and 4.7 g / kg gallic acid.

[0096] 7.3 Grapefruit-flavored beer mix (<0.1% alcohol by volume):

[0097] [Table 14] 1) The extract contains 10.3 g / kg of gallic acid.

[0098] 7.4 Non-alcoholic malt beverages (<0.1% alcohol by volume):

[0099] [Table 15] 1) The extract contains 10.3 g / kg of gallic acid.

[0100] 7.5 Tomato ketchup:

[0101] [Table 16] 1) The extract contains 2.2 g / kg of chlorogenic acid.

[0102] 7.6 Cream Dressing (approximately 55% water by weight):

[0103] [Table 17] 1) The extract contains 8.1 g / kg chlorogenic acid. Yet another aspect of the present invention may be as follows. [1] (A) Plants of the following families: Asparagaceae, Cannabaceae, Ericaceae, Lythraceae, Krameriaceae, Rosaceae, and Vitaceae; Preferably, the following plants are used: hops (Humulus lupulus), cultivated strawberry (Fragaria x anapassa), terrestris (Krameria lappacea), pomegranate (Punica granatum), lowbush blueberry (Vaccinium angustifolium), mountain cranesbill (Vaccinium macrocarpon), vine (Vaccinium myrtillus), cowberry (Vaccinium vitis-idaea), European grape (Vitis vinifera), The extract, Those fractions, one or more plant extracts and / or one or more fractions thereof having antibacterial effect selected from the group consisting of: (B) Abrusoside and Balansines, malic acid, benzaldehyde, gamma-decalactone, delta-decalactone, 3,7'-dihydroxy-4'-methoxyflavan isomers, eriodictyol, ethyl 2-methylbutyrate, ethyl butyrate, ethyl caproate, geranial, hesperetin, or extract of sweet tea (Rubus suavissimus), hesperetin dihydrochalcone, hesperidin dihydrochalcone, hydroxybenzoic acid amides, preferably 2,4-dihydroxybenzoic acid vanillylamide, 4-hydroxydihydrochalcones, preferably phloretin and davidiginin, p-hydroxybenzaldehyde, p-hydroxybenzoic acid, homoeriodictyol, limonene, lactic acid, menthofloractone, mogrosides, naringenin, naringin dihydrochalcone, neoisoflavonoids ide, neohesperidin dihydrochalcone, neral, δ-octalactone, pellitorin, piperonal, phloridodin, phyllodulcin isomers, rubusoside, a mixture of rubusoside isomers and homologues, 1-(2,4-dihydroxy-phenyl)-3-(3-hydroxy-4-methoxy-phenyl)-propan-1-one, stevioside and rebaudioside, trilobatin, vanillin, vanillic acid, vanillyl lignan, tartaric acid and cinnamon aldehyde, one or more flavoring agents which have an antimicrobial effect and / or which increase the antimicrobial efficacy of component (A) and / or which reduce, mask or modify the unpleasant taste impression of component (A) and / or which increase or modify the pleasant taste impression of component (A); 2. Use of a mixture comprising or consisting of as an antibacterial mixture, preferably a synergistic antibacterial mixture. [2] The use of the above-mentioned [1] as an antibacterial mixture, in which component (B) improves the antibacterial efficacy of component (A) and / or reduces, masks or modifies the unpleasant taste impression of component (A) and / or increases or modifies the pleasant taste impression of component (A). [3] Dekkera bruxellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomycodes ludwigii, Zygosaccharomyces bailii, and Pichia sp. Aureobasidium sp., Alternaria alternata, Aspergillus brasiliensis, Aspergillus fumigatus, Cladosporium sp., Fusarium sp., Rhodotorula sp., Sporidiobolus sp., Sporobolomyces sp., Paecilomyces sp. and Penicillium sp., The use according to [1] or [2] above as an antibacterial mixture, preferably as a synergistic antibacterial mixture, against one or more bacteria selected from the group consisting of: [4] The use according to any one of [1] to [3] above, wherein the extract of component (A) is selected from the group consisting of berry extracts, preferably red berry extracts, pomegranate extracts and herb extracts. [5] The use of any one of [1] to [4] above as an antibacterial mixture, preferably a synergistic antibacterial mixture, against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailiii, C. albicans, A. pullulans and A. braziliensis. [6] The use according to any one of [1] to [5], wherein component (B) contains or consists of at least one, two, three or more flavorings selected from the group consisting of eriodictyol, hesperetin, homoeriodictyol, naringenin, pellitoriin and vanillic acid. [7] The use according to any one of [1] to [6], wherein component (A) contains at least one, two, three or more compounds selected from the group consisting of catechin, chlorogenic acid, p-coumaric acid, epicatechin, ellagic acid, gallic acid, caffeic acid, sinapic acid and cinnamic acid. [8] The use according to any one of [1] to [7] above, wherein the ratio of the total amount of component (A) to the total amount of component (B) is in the range of 100:1 to 1:1, preferably 10:1 to 1:1. [9] A preparation for use for nutritional or enriched purposes or a pharmaceutical preparation comprising the mixture according to any one of [1] to [8] above, wherein the mixture is present in an amount sufficient to achieve an antibacterial effect, preferably a synergistic antibacterial effect.

[10] The mixture comprises: Dekkera brucellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomycodes ludwigii, Zygosaccharomyces bailii and Pichia spp. Aureobasidium spp., Alternaria alternata, Aspergillus braziliensis, Aspergillus fumigatus, Cladosporium spp., Fusarium spp., Rhodotorula spp., Sporidiobolus spp., Sporobolomyces spp., Paecilomyces spp., and Penicillium spp. The preparation according to claim 9, wherein the preparation contains an amount sufficient to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more bacteria selected from the group consisting of:

[11] The preparation described in [9] or

[10] , wherein the mixture is contained in an amount sufficient to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailii, C. albicans, A. pullulans and A. brasiliensis.

[12] The preparation according to any one of [9] to

[11] , wherein the preparation is selected from the group consisting of food products, preferably food products having ingredients of animal origin, such as mayonnaise, minced meat and sausage products, and sugar-containing food products, such as sweetened drinks, syrups, ketchup, cream dressings and dressings.

[13] (i) preparing a mixture according to any one of [1] to [8]; (ii) providing one or more further ingredients suitable for consumption; (iii) mixing the ingredients provided in steps (i) and (ii); A method for producing the preparation according to any one of claims 9 to 12, comprising or consisting of the steps:

[14] (i) preparing a mixture according to any one of [1] to [8]; (ii) providing the preparation to be treated, or one or more further components of the preparation to be treated, suitable for consumption; (iii) applying to the prepared preparation or to the components of the preparation suitable for consumption a mixture according to any one of [1] to [8]; To achieve an antibacterial effect, preferably to achieve a synergistic antibacterial effect, Preferably, Dekkera brucellensis, Candida albicans, Candida parapsilosis, Candida pseudointermedia, Saccharomyces cerevisiae, Saccharomycodes ludwigii, Zygosaccharomyces bailii and Pichia spp. Aureobasidium spp., Alternaria alternata, Aspergillus braziliensis, Aspergillus fumigatus, Cladosporium spp., Fusarium spp., Rhodotorula spp., Sporidiobolus spp., Sporobolomyces spp., Paecilomyces spp., and Penicillium spp. In order to achieve an antibacterial effect against one or more bacteria selected from the group consisting of, preferably, in order to achieve a synergistic antibacterial effect, More preferably, to achieve an antibacterial effect, preferably a synergistic antibacterial effect, against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailiii, C. albicans, A. pullulans and A. brasiliensis, adding each in sufficient quantity; 2. A method for the antimicrobial treatment of a food or a preparation used for pleasure or a pharmaceutical preparation, comprising or consisting of the steps:

Claims

1. 1. Use of component (A) grape seed extract of Vitis vulgare and component (B) vanillic acid as an antibacterial mixture against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailii, and C. albicans.

2. 2. The use according to claim 1, wherein the ratio of the total amount of component (A) to the total amount of component (B) is in the range of 100:1 to 1:

1.

3. A preparation comprising component (A) a grape seed extract of Vitis vulgare and component (B) vanillic acid for achieving an antibacterial effect against one or more or all of the bacteria selected from the group consisting of S. cerevisiae, Z. bailii, and C. albicans.

4. The preparation according to claim 3, wherein the preparation is a food product.

5. The preparation described in claim 3, wherein the preparation is a pharmaceutical preparation.

6. A method for producing the preparation according to any one of claims 3 to 5, comprising the steps of: (i) providing component (A) a grape seed extract of Vitis vulgare, and component (B) vanillic acid; (ii) providing one or more optional ingredients other than components (A) and (B) in said preparation; (iii) mixing the ingredients provided in steps (i) and (ii); A method comprising or consisting of the steps of:

7. A method for the antimicrobial treatment of a preparation selected from food and pharmaceutical preparations against one or more or all bacteria selected from the group consisting of S. cerevisiae, Z. bailii, and C. albicans, (i) providing component (A) a grape seed extract of Vitis vulgare, and component (B) vanillic acid; (ii) providing a preparation to be treated; (iii) adding component (A) and component (B) to the prepared preparation; A method comprising or consisting of the steps of:

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