Antimicrobial lactobacillus ferment

Abstract

The present invention pertains to a method for producing an antimicrobial composition preferably comprising piceol. Moreover, the invention relates to an antimicrobial composition comprising or consisting of piceol, preferably in an excess amount compared to picein, further plant-derived substances and optionally picein. The invention further relates to a cosmetic, personal care, household, home care, pet care or pharmaceutical product comprising said antimicrobial composition. Moreover, the invention relates to the use of a microorganism, preferably a lactobacillus species, for producing an antimicrobial composition comprising piceol from a composition, preferably a plant-extract, comprising picein. In addition, the invention relates to the use of shikimic acid for increasing the antimicrobial efficacy of piceol and / or to the use of piceol for increasing the antimicrobial efficacy of shikimic acid. Further embodiments and aspects of the invention are presented in the attached claims as well as in the description and examples below.

Description

[0001] Antimicrobial Lactobacillus Ferment

[0002] The present invention pertains to a method for producing an antimicrobial composition preferably comprising piceol. Moreover, the invention relates to an antimicrobial composition comprising or consisting of piceol, preferably in an excess amount compared to picein, further plant-derived substances and optionally picein. The invention further relates to a cosmetic, personal care, household, home care, pet care or pharmaceutical product comprising said antimicrobial composition. Moreover, the invention relates to the use of a microorganism, preferably a lactobacillus species, for producing an antimicrobial composition comprising piceol from a composition, preferably a plant-extract, comprising picein. In addition, the invention relates to the use of shikimic acid for increasing the antimicrobial efficacy of piceol and / or to the use of piceol for increasing the antimicrobial efficacy of shikimic acid. Further embodiments and aspects of the invention are presented in the attached claims as well as in the description and examples below.

[0003] Microbial contaminants can not only cause infections, skin irritations and allergic reactions, they also significantly decrease the shelf-life of personal care products. If such products are not sufficiently preserved, microbial contamination manifests in e.g. smell, discoloration, and visible mould growth. Thus, antimicrobial agents are added to personal care products. By means of various chemical and biological processes, microorganisms can be stopped from deteriorating product quality, thereby enhancing the shelf-life.

[0004] For decades antimicrobial agents for personal care products have largely been derived from crude oil sources since they are cheap and easy to produce.

[0005] EP0167286A1 , for instance, teaches a process for producing crude 4-HAP. It describes the production of hydroxy aromatic ketones such as 4-hydroxyacetophenone (4-HAP) by the Friedel-Crafts acylation of monocyclic phenolic reactants.

[0006] DE2653601 A1 discloses the hydrogen fluoride-catalyzed acylation of phenolic compounds such as phenol itself with an acyl halide such as acetyl chloride to form hydroxy aromatic ketones.

[0007] EP0069597 B1 teaches the preparation of p-phenoybenzoyl compounds by reacting diphenyl ether and an appropriate acyl compound such as acetic anhydride in the presence of hydrogen fluoride.

[0008] The educts of these processes are mainly of petrochemical origin. The produced antimicrobials are thus crude oil derived. Increasingly, however, consumers demand that products comprise exclusively “natural” antimicrobial agents. The choice of personal care products shall contribute to reaching a “circular economy”. As a result, natural raw materials for antimicrobial agents steadily gain momentum.

[0009] Despite being widely and commonly used, the terms “natural” and “sustainable” have not been defined officially in cosmetics until enacting ISO 16128 in 2016 and 2017. Having two separate parts, the first part (ISO 16128-1) defines the terms “natural” and “derived natural” ingredients. The second part (ISO 16128-2) offers a guideline to calculate the “naturalness” of cosmetic ingredients. To comply with ISO 16128-2, a value of at least 0.9 from a maximum of 1.0 shall be reached. This can be achieved if the ingredient is derived from a “sustainable” source. “Sustainable” in this context means a material whose usage does not lead to the depletion of a limited resource (e.g. petroleum or fossil fuel). Using such materials means the usage of a carbon source, which has existed only since recently and is renewable.

[0010] To distinguish between a petroleum based - and a product which is from a renewable source, the skilled person knows various techniques, including the evaluation of carbon- 12 / carbon- 13 and / or hydrogen-1 / hydrogen-2 ratios and radiocarbon dating. The ASTM D6866-16 method rtml) is described as a standard method for determining the biobased content of solid, liquid, and gaseous samples. Such methods comprise or consist of radiocarbon dating. Being an unstable isotope of carbon known as14C, radiocarbon emits beta particles until finally decaying to the more stable14N. Since crude oil derived products originate from carbon sources dating back millions of years, radiocarbon has decayed. Considering that the amount of radiocarbon in “sustainable” current plant sources is known, it can hence be calculated how much of the total organic carbon is petrochemically derived.

[0011] Besides being sustainable, antimicrobial agents must obviously fulfil a multitude of further requirements, e.g., remain effective for an extended period, be compatible with the final formulation of a personal care product and have broad spectrum antimicrobial activity. Thus, modern personal care products must fulfil a diverse profile of requirements driven by environmental and efficacy concerns. Finding suitable substances having the stated properties to a sufficient degree is challenging for a person skilled in the art since there is no clear relationship between the chemical structure of a substance and its biological activity regarding microorganisms. The availability of products achieving such standards is highly limited. In the state of the art, there is thus an ongoing and steadily increasing need for effective, multifunctional antimicrobial agents derived from sustainable, natural sources. One antimicrobial agent widely used in the cosmetic industry is 4-hydroxyacetophenone (4- HAP), which is also called “piceol”. Derivable from its inactive precursor “picein,” the compound occurs in several plant genera such as the needles of Norway spruces (Picea abies). 4-HAP is described by the CAS number 99-93-4 (as listed by the European Chemicals Agency) and corresponds to the following structure:

[0012] 4-hydroxyacetophenone

[0013] 4 - HAP has many applications, for instance the stabilisation of pharmaceutical and cosmetic compositions. The compound - in this context known as p-hydroxyacetophenone - has potent antioxidant effects and can neutralize a multitude of free radicals. As a result, in cosmetics, hydroxyacetophenone has a boosting effect on the preservation system allowing cosmetic chemists to use lower amounts of classical preservatives such as phenoxyethanol. Consequently, preservative efficacy can be kept while at the same time reducing the risk of allergic reactions. Moreover, hydroxyacetophenone is stable across a wide pH-range and types of emulsions, giving it great versatility.

[0014] Typically, when 4-hydroxyacetophenone is produced on an industrial scale, the compound is obtained by chemical synthesis. Briefly, phenol reacts with acetic acid or acetic anhydride in the presence of water-free hydrogen fluoride (HF) or hydrofluoric acid to form phenyl acetate, which is transformed to 4-hydroxyacetophenone by Fries rearrangement. However, hydrogen fluoride and hydrofluoric acid are toxic substances. Thus, chemical production processes require complex safety mechanisms, special equipment, and specific conditions for waste disposal. Hence, natural, biotechnological production methods are of increasing interest. Unfortunately, the production of antimicrobial compositions from natural sources is challenging due to a low yield.

[0015] A first, primary object of the present invention was thus to provide an environmentally friendly method for producing an antimicrobial composition from exclusively renewable sources, preferably with enhanced antimicrobial activity. A further object of the invention was to provide a natural, effective antimicrobial composition. Moreover, it was an object of the invention to improve the antimicrobial properties of natural plant extracts. An additional object of the present invention was to provide new combinations of antimicrobial agents leading to a synergistic improvement of the antimicrobial activity of the resulting mixture. Additional objects of the present invention can be derived from the attached claims and the description below.

[0016] The primary object of the invention is surprisingly achieved by the method according to claim 1. Preferable embodiments are subject of the dependent claims, the description below, and the figures.

[0017] In a first aspect, the invention relates to a method for producing an antimicrobial composition, preferably comprising piceol, comprising the following steps:

[0018] -providing a growth medium comprising picein and at least one microorganism

[0019] -fermenting the mixture comprising picein and said microorganism, thereby forming the antimicrobial composition comprising piceol

[0020] -optionally removing the cells of the microorganism from the mixture

[0021] -and / or optionally inactivating the microorganism

[0022] Surprisingly, it was found that the method according to the invention forms a natural antimicrobial composition with a significantly enhanced yield of piceol. In turn, the concentration of its less effective precursor, picein, is significantly decreased compared to its concentration in the natural, untreated plant extract. The invention thus constitutes an environmentally friendly alternative to producing antimicrobial agents by conventional, crude oil-based means. In the context of the present invention, a composition is preferably defined as “antimicrobial composition” if it contains piceol.

[0023] In a preferred embodiment, the method provides a piceol content of at least 100 ppm, preferably 200 ppm, further preferably of at least 300 ppm, more preferably of at least 500 ppm, even more preferably of at least 1000 ppm, and most preferably of at least 3000 ppm piceol.

[0024] In a further preferred embodiment, the invention provides an antimicrobial composition with a weight ratio of piceol to picein of at least 2: 1 , more preferred of at least 4: 1 , even more preferred of at least 10: 1 , further preferred 40: 1 and most preferred of at least 100: 1.

[0025] In yet a further preferred embodiment, the weight percentage of piceol with respect to the summed weight of piceol and picein in the antimicrobial composition amounts to at least 50 %, preferably 75 %, more preferably to at least 95 wt%, and even more preferably to at least 97wt%. Most preferably 100 wt% piceol is produced.

[0026] Preferably, the picein is a picein extract, preferably extracted from a plant selected from the group consisting of Spruce, Salix, Phagnalon Rupestre, Rhodiola Rosea, Poacynum hendersonii, Baccharis magellanica or Vauquelinia. Such plant extracts contain picein and piceol. Picein, the precursor of piceol showing less antimicrobial efficacy, is present in untreated extracts to a higher degree than piceol. In a preferred embodiment, the picein extract is extracted from a Spruce needle and / or a Baccharis magellanica plant.

[0027] In a further preferred embodiment, the method comprises that the picein extracts are extracted using a polar solvent, i.e. a solvent with a dielectric constant greater than 15. Preferably, the picein extraction is carried out with a polar solvent selected from the group consisting of ethanol, water, methanol, isopropanol and mixtures of two or more of these solvents. Most preferably, picein is extracted by water extraction leading to a transparent antimicrobial composition, which is completely natural and sustainable.

[0028] Preferred general extraction processes are maceration, re-maceration, digestion, agitation maceration, vortex extraction, ultrasonic extraction, counter current extraction, percolation, repercolation, evacolation (extraction under reduced pressure), subcritical or supercritical fluid extraction, diacolation and solid / liquid extraction under continuous reflux. Percolation is even more preferred and has good upscaling properties. Following the extraction process, the crude extracts obtained may optionally be subjected to other typical steps, such as, for instance, fractionation, purification and / or decoloration.

[0029] Lactic acid bacteria (LAB) have found widespread usage as probiotics and various health- related applications, i.e. the irritable bowel syndrome (IBS). Preferably, the microorganism in the method according to the invention is a Lactobacillus species, preferably selected from the group consisting of Lactobacillus acetotolerans, Lactobacillus acidophilus, Lactobacillus agrestimuris, Lactobacillus alvei, Lactobacillus amylolyticus, Lactobacillus amylovorus, Lactobacillus animate, Lactobacillus apis, Lactobacillus bombicola, Lactobacillus brevis, Lactobacillus brevisimilis, Lactobacillus casei, Lactobacillus catenefornis, Lactobacillus colini, Lactobacillus corticis, Lactobacillus crispatus, Lactobacillus delbrueckii, Lactobacillus equicursoris, Lactobacillus faeni, Lactobacillus fermentum, Lactobacillus fornicalis, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus gigeriorum, Lactobacillus guizhouensis, Lactobacillus halophilus, Lactobacillus hamsteri, Lactobacillus helsingborgensis, Lactobacillus helveticus, Lactobacillus hominis, Lactobacillus huangpiensis, Lactobacillus iatae, Lactobacillus iners, Lactobacillus insectis, Lactobacillus intermedius, Lactobacillus intestinalis, Lactobacillus isalae, Lactobacillus japonicus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus juensis, Lactobacillus kalixensis, Lactobacillus kefiranofaciens, Lactobacillus kimbladii, Lactobacillus kitasatonis, Lactobacillus kullabergensis, Lactobacillus laiwuensis, Lactobacillus larvae, Lactobacillus leichmannii, Lactobacillus letivazi, Lactobacillus melliventris, Lactobacillus mobilis, Lactobacillus mulieris, Lactobacillus nasalidis, Lactobacillus panisapium, Lactobacillus paracasei, Lactobacillus paragasseri, Lactobacillus paraplantarum, Lactobacillus pasteurii, Lactobacillus plantarum, Lactobacillus porci, Lactobacillus psittaci, Lactobacillus rennanquilfy, Lactobacillus reuteri, Lactobacillus rizhaonensis, Lactobacillus rhamnosus, Lactobacillus rodentium, Lactobacillus rogosae, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus selangorensis, Lactobacillus sucicola, Lactobacillus taiwanensis, Lactobacillus terrae, Lactobacillus ultunensis, Lactobacillus vermiforme, Lactobacillus xujianguonis and Lactobacillus xylocopicola.

[0030] In a further preferred embodiment, the microorganism is selected from the group consisting of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus paraplantarum, Lactobacillus reuteri, Lactobacillus pentosus, Lactobacillus buchneri, Lactobacillus sucicola, Lactobacillus bombi, Lactobacillus gasseri and Lactobacillus rodentium.

[0031] In yet another preferred embodiment, the microorganism is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum.

[0032] In another preferred embodiment, the invention provides a piceol content of at least 100 ppm after a brief fermentation time. Further preferably, the invention provides a piceol content of at least 100 ppm after 2h of incubation with a Lactobacillus species. Preferably, the Lactobacillus species is a Lactobacillus rhamnosus.

[0033] In a further preferred embodiment, the lactobacillus species is grown in a growth medium, preferably in a bacterial growth medium and more preferably in a Man-Rogosa-Sharpe (MRS) medium, or any MRS based medium for optimized fermentation conditions. The picein extract is added to the Lactobacillus fermentation. This can be done at the beginning of the fermentation, during the exponential growth phase, or at the stationary phase. Subsequently, in a preferred embodiment, the Lactobacillus cells are grown until exponential growth phase, followed by the complete or partial replacement of the growth medium by a simplified medium, preferably without glucose, for the actual digestion of the picein extract. In a preferred variant, the cell density is adjusted in this step to an optical density (OD620) of 5 or higher.

[0034] In a further preferred embodiment, the picein from the plant material is directly extracted and fermented within the Lactobacillus fermentation medium without a physical separation of both steps.

[0035] In yet another preferred embodiment, the method for producing an antimicrobial composition, preferably comprising piceol, comprises the following steps:

[0036] -providing a growth medium comprising picein and at least one microorganism

[0037] -fermenting the mixture comprising picein and said microorganism thereby forming the antimicrobial composition, preferably comprising piceol

[0038] -optionally removing the cells of the microorganism from the mixture by centrifugation, filtration or cell lysis,

[0039] -and / or optionally inactivating said microorganism by a heat-treatment, preferably at a temperature in the range of 60 to 121 °C for 1 second to 120 minutes.

[0040] Advantageously, said heat-treatment conditions lead to a complete inactivation of the microorganism.

[0041] In yet a further preferred embodiment, the picein extract is a Spruce needle extract and / or a Baccharis magnellanica extract, and the Lactobacillus is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum.

[0042] In still another preferred embodiment, the picein extract is a Spruce needle extract and / or an extract of the plant Baccharis magellanica and the microorganism according to the invention is a Lactobacillus rhamnosus strain.

[0043] In a further preferred embodiment, the method comprises the step of downstream processing of the formed antimicrobial composition, preferably wherein the downstream processing comprises a method selected from the group consisting of centrifugation, crystallization, solidphase adsorption, filtration, reverse osmosis, precipitation, decantation, liquid / liquid extraction, vacuum distillation, falling film distillation, melt crystallization, drying and / or combinations thereof. In a preferred embodiment, the microorganisms are filtered out. In a further preferred embodiment, heat-inactivation, e.g. by tyndallization or pasteurization, or cell lysis is used as a means for inactivating said microorganisms. Further preferably, the antimicrobial composition is recovered from said mixture.

[0044] In a preferred embodiment, the antimicrobial composition formed according to the method of the invention is a liquid concentrate or a solid concentrate, such as a powder. The concentrate is produced after extraction and optional separation of the biomass from the antimicrobial composition either without or with prior partially removal of the liquid and after optional addition of a solid carrier such as e.g. modified starches like maltodextrin, dextrin or cyclodextrin, lactose, modified celluloses, gums like xanthan gum, gellan gum, guar gum, gum arabic, gum ghatti, tragacanth gum or locust bean gum, silicium dioxide, preferably maltodextrin or mixtures of two or more of these by drying using suitable processes such as spray , freeze- or vacuum drying.

[0045] In a further preferred embodiment, the antimicrobial composition formed according to the invention is a concentrate comprising or consisting of: a) 0.5 to 80 wt.% piceol according to the invention b) 0.5 to 90 wt.% water, and / or c) 0.5 to 90 wt.% carrier.

[0046] The weight ratios are calculated based on piceol dry weight. Even more preferred is a content of 0.5 to 40 wt.% piceol. In a further preferred embodiment, a content of 10 to 80 wt.% water is employed. Additionally, a content of 10 to 80 wt.% carrier is preferred. If the concentrate is a liquid concentrate, it advantageously comprises 1 to 70 wt.% water, preferably 10 to 60 wt.% water.

[0047] If the antimicrobial composition is a solid concentrate, it preferably comprises or consists of a) 0.5 to 10 wt.% piceol according to the invention, b) 0.5 to 8 wt.% water, and c) 15 to 98 wt.% solid carrier, preferably maltodextrin. The weight ratios are calculated based on piceol dry weight.

[0048] In yet another preferred embodiment, the above concentrate further comprises 0.1 to 5 wt.% of one or more preservatives or a preservative system. In another preferred form, the concentrate comprises also stabilizers.

[0049] Even more preferred is the use of 0.5 to 2 wt.% of one or more preservatives or preservative systems or stabilizers. The amount of the respective components is chosen in compliance with the Cosmetics Directive 76 / 768 / EEC and Ell Directive 95 / 17 / EC. Preferably the preservatives are employed according to the classes and compounds listed in the Appendix 6, Parts A and B of the Cosmetics Directive 76 / 768 / EEC. Exemplary preservatives are benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, phenoxyethanol, or combinations thereof. Preservative boosters are preferably hydroxyacetophenone, 1 ,2-pentanediol, 1 ,2-hexanediol,

[0050] 1 .2-octanediol or combinations thereof. However, 1 ,2-pentanediol may also be used in higher amounts as a secondary liquid carrier.

[0051] Alternatively, further substances may be added before drying, such as polyols e.g. glycerin,

[0052] 1.2-propanediol (propylene glycol), 1 ,3-propanediol, 1 ,2-butanediol, 1 ,3-butanediol (butylene glycol), 1 ,2-pentylene glycol and 1 ,2-hexanediol or mixtures of two or more these. In such cases, typically an aqueous solvent system is achieved, with the active components dissolved therein.

[0053] Even more preferably, the antimicrobial composition formed according to the method of the invention is a liquid concentrate comprising or consisting of: a) 0.5 to 10 wt.% piceol according to the invention, b) 1 to 70 wt.% water, c) 0.5 to 85 wt.% liquid carrier, and d) optionally 0.1 to 5 wt.% of a preservative or preservative system.

[0054] The weight ratios are calculated based on piceol dry weight. Advantageously, it was found that the antimicrobial composition, both as liquid or solid concentrate, shows good storage properties, is easy to handle, dose and formulate.

[0055] In a further preferred embodiment, the formed antimicrobial composition is substantially non- petrochemical. Said composition is particularly advantageous due to its environmental friendliness and its naturalness, thereby meeting increasing consumer needs and complying with ISO 16128. The composition is considered “substantially non-petrochemical”, if at least 75 %, preferably at least 80 %, further preferably at least 85 %, particularly preferably at least 90 %, even further preferably at least 95 % of its carbon, based on the total carbon content in the composition, is non-petrochemically-derived carbon. Preferably, at least 75 %, preferably at least 80 %, further preferably at least 85 %, particularly preferably at least 90 %, and even further preferably at least 95 % of the carbon of the piceol in the composition is non- petrochemically-derived carbon. Most preferably, all carbon of piceol in the composition is non- petrochemically-derived carbon.

[0056] To distinguish between a petroleum based - and a product which is from a renewable source (“non-petrochemical”), the skilled person knows various techniques, including the evaluation of carbon- 12 / carbon- 13 and / or hydrogen-1 / hydrogen-2 ratios and radiocarbon dating. For instance, the ASTM D6866-16 method ( ) is described as a standard method for determining the biobased content of solid, liquid, and gaseous samples. Such methods comprise or consist of radiocarbon dating. Being an unstable isotope of carbon known as14C, radiocarbon emits beta particles until finally decaying to the more stable14N. Since crude oil derived products originate from carbon sources dating back millions of years, radiocarbon has decayed. Considering that the amount of radiocarbon in “sustainable” current plant sources is known, it can hence be calculated how much of the total organic carbon is petrochemically derived.

[0057] The above liquid or solid antimicrobial composition can be employed in cosmetic and / or dermatological and / or pharmaceutical products for skin and hair care and cleansing in an an amount of 0.001 to 10 wt.%, preferably 0.01 to 5 wt.%, and more preferably 0.1 to 2 wt.%.

[0058] In yet another preferred embodiment, the growth medium for producing an antimicrobial composition does not comprise Escherichia coli (E. coli). Preferably, the fermentation of the plant extract is not performed by E. coli. Further preferably, the growth medium for producing antimicrobial composition does not comprise a genetically modified organism (GMO). In another aspect, the invention relates to an antimicrobial composition, preferably comprising piceol, obtained or obtainable by a method comprising or consisting of the steps as defined in any of the method claims.

[0059] It was found that using the method according to the invention provides piceol from non- petrochemically sources. Hence, the produced piceol is also non-petrochemically-derived. The natural antimicrobial composition obtained by the method comprises a significantly enhanced yield of piceol. It was further found that using the method according to the invention provides an antimicrobial composition that comprises piceol, at least one lactobacillus species and organic acids, preferably shikimic acid. This combination offers a variety of cosmetic benefits including a strengthening of the skin barrier function, a reduction of transepidermal water loss, and an increase of components of the natural moisturizing factor (NMF). Moreover, the antimicrobial composition obtained or obtainable by the method according to the invention supports in maintaining, establishing and restoring a healthy state of the skin microbiome. Furthermore, said antimicrobial composition promotes skin cell turnover, improves the skin's appearance and texture, while at the same time having antimicrobial efficacy to protect the skin and / or the cosmetic formulation.

[0060] In another aspect, the invention relates to an antimicrobial composition comprising or consisting of piceol, preferably in an excess amount compared to picein, further plant-derived substances and optionally picein.

[0061] In a preferred embodiment, said further plant-derived substances are selected from the group consisting of organic acids such as shikimic acid, quinic acid, citric acid, malic acid, succinic acid, threonic acid and lactic acid.

[0062] In a preferred embodiment, the antimicrobial composition comprises or consists of picein, piceol, wherein piceol is an excess of picein and optionally further plant-derived substances.

[0063] In another preferred embodiment, the antimicrobial composition comprises piceol, preferably in an excess amount compared to picein, shikimic acid and optionally picein.

[0064] Surprisingly, it was found that the antimicrobial composition according to the invention shows a synergistically intensified antimicrobial effect, which is superior to the components piceol and organic acids such as shikimic acid measured individually. Particularly regarding Staphylococcus aureus (S. aureus), a more than additive antimicrobial reaction was observed for an antimicrobial composition comprising piceol and shikimic acid. Organic acids such as shikimic acid, quinic acid, citric acid, malic acid, succinic acid, threonic acid and lactic acid offer a variety of cosmetic benefits, primarily related to their exfoliating efficacy. By promoting skin cell turnover, they help to improve skin's appearance and texture.

[0065] Shikimic acid, a cyclohexene, listed under the CAS number 138-59-0, is a naturally occurring organic acid and corresponds to the following structure:

[0066] Shikimic acid

[0067] Shikimic acid can be found in many plants such as pine needles and I llicium verum.

[0068] In a further preferred embodiment, the ratio of the total weight of piceol in the composition to the total weight of shikimic acid in the composition is from 5:1 to 1 :20, further preferably from 2:1 to 1 :15, particularly preferably from 1:1 to 1 :10 and most preferably from 1 :3 to 1:5.

[0069] In yet another preferred embodiment, the antimicrobial composition additionally comprises at least one microorganism, preferably a Lactobacillus species.

[0070] In a preferred embodiment, the lactobacillus is selected from the list consisting of Lactobacillus acetotolerans, Lactobacillus acidophilus, Lactobacillus agrestimuris, Lactobacillus alvei, Lactobacillus amylolyticus, Lactobacillus amylovorus, Lactobacillus animate, Lactobacillus apis, Lactobacillus bombicola, Lactobacillus brevis, Lactobacillus brevisimilis, Lactobacillus casei, Lactobacillus catenefornis, Lactobacillus colini, Lactobacillus corticis, Lactobacillus crispatus, Lactobacillus delbrueckii, Lactobacillus equicursoris, Lactobacillus faeni, Lactobacillus fermentum, Lactobacillus fornicalis, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus gigeriorum, Lactobacillus guizhouensis, Lactobacillus halophilus, Lactobacillus hamsteri, Lactobacillus helsingborgensis, Lactobacillus helveticus, Lactobacillus hominis, Lactobacillus huangpiensis, Lactobacillus iatae, Lactobacillus iners, Lactobacillus insectis, Lactobacillus intermedius, Lactobacillus intestinalis, Lactobacillus isalae, Lactobacillus japonicus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus juensis, Lactobacillus kalixensis, Lactobacillus kefiranofaciens, Lactobacillus kimbladii, Lactobacillus kitasatonis, Lactobacillus kullabergensis, Lactobacillus laiwuensis, Lactobacillus larvae, Lactobacillus leichmannii, Lactobacillus letivazi, Lactobacillus melliventris, Lactobacillus mobilis, Lactobacillus mulieris, Lactobacillus nasalidis, Lactobacillus panisapium, Lactobacillus paracasei, Lactobacillus paragasseri, Lactobacillus paraplantarum, Lactobacillus pasteurii, Lactobacillus plantarum, Lactobacillus porci, Lactobacillus psittaci, Lactobacillus rennanquilfy, Lactobacillus reuteri, Lactobacillus rizhaonensis, Lactobacillus rhamnosus, Lactobacillus rodentium, Lactobacillus rogosae, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus selangorensis, Lactobacillus sucicola, Lactobacillus taiwanensis, Lactobacillus terrae, Lactobacillus ultunensis, Lactobacillus vermiforme, Lactobacillus xujianguonis and Lactobacillus xylocopicola.

[0071] In a further preferred embodiment, the lactobacillus is selected from the list consisting of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus paraplantarum, Lactobacillus reuteri, Lactobacillus pentosus, Lactobacillus buchneri, Lactobacillus sucicola, Lactobacillus bombi, Lactobacillus gasseri and Lactobacillus rodentium.

[0072] In a most preferred embodiment, the microorganism is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum.

[0073] In another preferred embodiment, the antimicrobial composition comprises or consists of at least one microorganism, preferably a Lactobacillus species, piceol, preferably in an excess amount compared to picein, optionally picein and optionally further plant-derived substances. Advantageously, the presence of a Lactobacillus species in the antimicrobial composition leads to a multitude of skin-related benefits, e.g. a strengthening of the skin barrier function, a reduction of transepidermal water loss and an increase of components of the natural moisturizing factor (NMF). Moreover, the Lactobacillus species in the antimicrobial composition supports in maintaining, establishing and restoring a healthy state of the skin microbiome. In yet another preferred embodiment, the antimicrobial composition according to the invention comprises a piceol content of at least 100 ppm, preferably 200 ppm, further preferably of at least 300 ppm, more preferably of at least 500 ppm, even more preferably of at least 1000 ppm, and most preferably of at least 3000 ppm piceol.

[0074] In a further preferred embodiment, the invention provides an antimicrobial composition comprising piceol in an excess amount compared to picein, preferably wherein the excess is a weight ratio of piceol to picein of at least 2 : 1 , more preferred of at least 4 : 1 , even more preferred of at least 10 : 1 , further preferred 40:1 and most preferred of at least 100 : 1 .

[0075] In yet a further preferred embodiment, the weight percentage of piceol with respect to the summed weight of piceol and picein in the antimicrobial composition amounts to at least 50%, preferably to at least 75 %, more preferably to at least 95 wt% and even more preferably to at least 97 wt%. Most preferably, 100 wt% piceol is produced from picein.

[0076] In another preferred embodiment, the invention provides a piceol content of at least 100 ppm after a brief fermentation time. Preferably, the invention provides a piceol content of at least 100 ppm after 2h of incubation with a Lactobacillus species. Preferably, the Lactobacillus species is a Lactobacillus rhamnosus.

[0077] In a preferred embodiment, the composition comprises at least 75 wt%, preferably at least 80 wt%, further preferably at least 85 wt%, particularly preferably at least 90 wt%, even further preferably at least 95 wt% non-petrochemically-derived carbon based on the total carbon content of the piceol in the composition. Most preferably, all carbon of the piceol in the composition is non-petrochemically-derived carbon based on the total carbon content of the piceol in the composition.

[0078] Preferably, the antimicrobial composition comprises piceol, shikimic acid a Lactobacillus species, preferably selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, Lactobacillus plantarum, and optionally picein.

[0079] In a preferred embodiment, the antimicrobial composition according to the invention is a liquid concentrate or a solid concentrate, such as a powder. Preferably, the concentrate is produced after extraction and optional separation of the biomass from the antimicrobial composition either without or with prior partially removal of the liquid and after optional addition of a solid carrier such as e.g. modified starches like maltodextrin, dextrin or cyclodextrin, lactose, modified celluloses, gums like xanthan gum, gellan gum, guar gum, gum arabic, gum ghatti, tragacanth gum or locust bean gum, silicium dioxide, preferably maltodextrin or mixtures of two or more of these by drying using suitable processes such as spray , freeze- or vacuum drying.

[0080] In a further preferred embodiment, the antimicrobial composition formed according to the invention is a concentrate comprising or consisting of: a) 0.5 to 80 wt.% piceol according to the invention b) 0.5 to 90 wt.% water, and / or c) 0.5 to 90 wt.% carrier.

[0081] The weight ratios are calculated based on piceol dry weight. Even more preferred is a content of 0.5 to 40 wt.% piceol. In a further preferred embodiment, a content of 10 to 80 wt.% water is employed. Additionally, a content of 10 to 80 wt.% carrier is preferred. If the concentrate is a liquid concentrate, it advantageously comprises 1 to 70 wt.% water, preferably 10 to 60 wt.% water.

[0082] If the antimicrobial composition is a solid concentrate, it preferably comprises or consists of a) 0.5 to 10 wt.% piceol according to the invention, b) 0.5 to 8 wt.% water, and c) 15 to 98 wt.% solid carrier, preferably maltodextrin.

[0083] The weight ratios are calculated based on piceol dry weight.

[0084] In yet another preferred embodiment, the above concentrate further comprises 0.1 to 5 wt.% of one or more preservatives or a preservative system. In another preferred form, the concentrate comprises also stabilizers.

[0085] Even more preferred is the use of 0.5 to 2 wt.% of one or more preservatives or preservative systems or stabilizers. The amount of the respective components is chosen in compliance with the Cosmetics Directive 76 / 768 / EEC and Ell Directive 95 / 17 / EC. Preferably, the preservatives are employed according to the classes and compounds listed in the Appendix 6, Parts A and B of the Cosmetics Directive 76 / 768 / EEC. Exemplary preservatives are benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, phenoxyethanol, or combinations thereof. Preservative boosters are preferably hydroxyacetophenone, 1 ,2-pentanediol, 1 ,2-hexanediol,

[0086] 1 .2-octanediol or combinations thereof. However, 1 ,2-pentanediol may also be used in higher amounts as a secondary liquid carrier.

[0087] Alternatively, further substances may be added before drying, such as polyols e.g. glycerin,

[0088] 1.2-propanediol (propylene glycol), 1 ,3-propanediol, 1 ,2-butanediol, 1 ,3-butanediol (butylene glycol), 1 ,2-pentylene glycol and 1 ,2-hexanediol or mixtures of two or more these. In such cases, typically an aqueous solvent system is achieved, with the active components dissolved therein.

[0089] Even more preferably, the antimicrobial composition formed according to the method of the invention is a liquid concentrate comprising or consisting of: a) 0.5 to 10 wt.% piceol according to the invention, b) 1 to 70 wt.% water, c) 0.5 to 85 wt.% liquid carrier, and d) optionally 0.1 to 5 wt.% of a preservative or preservative system.

[0090] The weight ratios are calculated based on piceol dry weight.

[0091] Advantageously, it was found that the antimicrobial composition, both as liquid or solid concentrate, shows good storage properties, is easy to handle, dose, and formulate.

[0092] In yet another aspect, the invention relates to a cosmetic, personal care, household, home care, pet care or pharmaceutical product comprising the antimicrobial composition according to the invention.

[0093] Such products preferably comprise the antimicrobial composition according to the invention in the product in an amount of 0.001 to 10 wt.%, preferably 0.01 to 5 wt.%, more preferably 0.1 to 2 wt.%. Cosmetic, personal care, household, home care, pet care or pharmaceutical products, as described herein, are particularly products which are suitable for topical application to the skin, in particular to achieve a cosmetic effect.

[0094] Preferably, the cosmetic or personal care product according to the present invention is selected from the group of products for treatment, protecting, care and cleansing of the skin and / or hair or as a make-up product, preferably as a leave-on product, more preferably in the form of or selected from the product group consisting of alcoholic or aqueous / alcoholic solution, dispersion, suspension, emulsion (preferably cream, lotion or milk of the W / O, O / W or multiple emulsion, PIT emulsion, emulsion foam, micro-, nanoemulsion, Pickering emulsion type), ointment, paste, gel (preferably hydro-, hydrodispersion-, oleogel), balm, serum, powder, wipe, Eau de Toilette, Eau de Cologne, perfume, stick, roll-on, (pump) spray, aerosol, leave-on skin care composition (preferably face-care composition), leave-on insect repellent composition, sunscreen composition, skin-lightening composition, self-tanning composition, aftersun preparation, shaving or after-shave composition, hair-removing com-position, hair care composition, preferably conditioner, hair lotion, hair tonic, styling cream, pomade, styling aid (preferably gel or wax), permanent wave and fixing compositions, hair smoothing composition (straightening composition, relaxer), hair setting composition, blonding composition, hair coloring composition, such as e.g. temporary, directly absorbed, semi-permanent hair coloring composition, permanent hair coloring composition, decorative cosmetic composition (preferably face powder, eye shadow, kajal pencil, lip-stick), deodorant and / or antiperspirant composition.

[0095] Preferably, pharmaceutical products according to the invention are in the form of capsules, tablets (uncoated and coated tablets, for example having coatings resistant to gastric juices), sugar-coated tablets, granules, pellets, mixtures of solids, dispersions in liquid phases, as emulsions, as powders, as solutions, as pastes or as other swallowable or chewable products and are preferably used as medicines only available by prescription, from pharmacies or other medicines or as food supplements.

[0096] Further preferably, the dosage form of a cosmetic product or a pharmaceutical product is selected from the group consisting of ointments, creams, pastes, gels, suspensions, crystal suspensions, emulsions, solutions, drops, powders, sprays, shampoos and foams.

[0097] In a further preferred embodiment, the product is selected from the group consisting of an oil- in-water or a water-in-oil emulsion, ointment, creme, lotion, and gel. More preferably, the cosmetic or pharmaceutical product is selected from the group comprising or consisting of soap, facial cleanser, exfoliant, mouthwash, toothpaste, hair wash, body wash, hand wash, essence, serum, toner, moisturizer, face mask, multipurpose cleaner, skin cream, lotion, deodorant.

[0098] The composition according to the present invention can advantageously be combined with other cosmetically or pharmaceutically active agents and / or adjuvants and / or additives or auxiliaries, such as are customarily used in such preparations or formulations, such as for example abrasives, anti-acne agents, agents against ageing of the skin, anti-cellulitis agents, anti-dandruff agents, anti-inflammatory agents, irritation-preventing agents, irritation-inhibiting agents, antioxidants, astringents, odour absorbers, perspiration-inhibiting agents, antiseptic agents, anti-statics, binders, buffers, carrier materials, chelating agents, cell stimulants, cleansing agents, depilatory agents, surface-active substances, deodorizing agents, antiperspirants, softeners, emulsifiers, enzymes, enzyme inhibitors, essential oils, fibres, filmforming agents, fixatives, foam-forming agents, foam stabilizers, substances for preventing foaming, foam boosters, gelling agents, gel-forming agents, hair care agents, hair-setting agents, hair-straightening agents, moisture-donating agents, moisturizing substances, moisture-retaining substances, bleaching agents, strengthening agents, stain-removing agents, optically brightening agents, impregnating agents, dirt-repellent agents, dyes, frictionreducing agents, lubricants, moisturizing creams, ointments, opacifying agents, plasticizing agents, covering agents, polish, preservatives, gloss agents, green and synthetic polymers, powders, proteins, reoiling agents, abrading agents, silicones, skin-soothing agents, skincleansing agents, skin care agents, skin-healing agents, skin-lightening agents, skin-protecting agents, skin-softening agents, hair promotion agents, cooling agents, skin-cooling agents, warming agents, skin-warming agents, stabilizers, surfactants, UV-absorbing agents, UV filters, primary sun protection factors, secondary sun protection factors, detergents, fabric conditioning agents, suspending agents, skin-tanning agents, actives modulating skin or hair pigmentation, matrix-metalloproteinase inhibitors, skin moisturizing agents, glycosaminoglycan stimulators, TRPV1 antagonists, desquamating agents, anti-cellulite agents or fat enhancing agents, hair growth activators or inhibitors, thickeners, rheology additives, vitamins, oils, waxes, pearlizing waxes, fats, phospholipids, saturated fatty acids, mono- or polyunsaturated fatty acids, a-hydroxy acids, polyhydroxy fatty acids, liquefiers, dyestuffs, colour protecting agents, pigments, anti-corrosives, fragrances or perfume oils, aromas, flavouring substances, odoriferous substances, polyols, electrolytes, organic solvents, and mixtures of two or more of the aforementioned substances, as further described below. Of the above cosmetically or pharmaceutically active agents and / or adjuvants and / or additives or auxiliaries agents against ageing of the skin, antioxidants, chelating agents, emulsifiers, surfactants, green and synthetic polymers, skin-cooling agents, rheology additives, oils, fragrances or perfume oils, and polyols are particularly preferred in the preparation of cosmetic, personal care, household, home care, pet care or pharmaceutical composition.

[0099] Further preferred embodiments and features described above apply accordingly to the cosmetic, personal care, household, home care, pet care or pharmaceutical product comprising the antimicrobial composition according to the invention.

[0100] In another aspect, the invention relates to the use of a microorganism, preferably a Lactobacillus species, for producing an antimicrobial composition comprising piceol from a composition, preferably a plant extract, comprising picein.

[0101] Surprisingly, it was found that the use of the microorganism according to the invention forms a natural antimicrobial composition with a significantly enhanced yield of piceol. In turn, the concentration of its less effective precursor, picein, is significantly decreased compared to its concentration in the natural, untreated plant extract. The invention thus constitutes an environmentally friendly alternative to producing antimicrobial agents by conventional, crude oil-based means. In the context of the present invention, a composition is preferably defined as “antimicrobial composition” if it contains piceol.

[0102] In a preferred embodiment, the use of the microorganism provides a piceol content of at least 100 ppm, preferably 200 ppm, further preferably of at least 300 ppm, more preferably of at least 500 ppm, even more preferably of at least 1000 ppm, and most preferably of at least 3000 ppm piceol.

[0103] In a further preferred embodiment, the invention provides an antimicrobial composition with a weight ratio of piceol to picein of at least 2: 1 , more preferred of at least 4: 1 , even more preferred of at least 10: 1 , further preferred 40: 1 and most preferred of at least 100: 1.

[0104] In yet a further preferred embodiment, the weight percentage of piceol with respect to the summed weight of piceol and picein in the antimicrobial composition amounts to at least 50 %, preferably 75 %, more preferably to at least 95 wt%, and even more preferably to at least 97wt%. Most preferably 100 wt% piceol is produced. Preferably, the picein is a picein extract, preferably extracted from a plant selected from the group consisting of Spruce, Salix, Phagnalon Rupestre, Rhodiola Rosea, Poacynum hendersonii, Baccharis magellanica or Vauquelinia. Such plant extracts contain picein and piceol. Picein, the precursor of piceol showing less antimicrobial efficacy, is present in untreated extracts to a higher degree than piceol. In a preferred embodiment, the picein extract is extracted from a Spruce needle and / or a Baccharis magellanica plant.

[0105] In a further preferred embodiment, the use comprises that the picein extracts are extracted using a polar solvent, i.e. a solvent with a dielectric constant greater than 15. Preferably, the picein extraction is carried out with a polar solvent selected from the group consisting of ethanol, water, methanol, isopropanol and mixtures of two or more of these solvents. Most preferably, picein is extracted by water extraction leading to a transparent antimicrobial composition, which is completely natural and sustainable.

[0106] Preferred general extraction processes are maceration, re-maceration, digestion, agitation maceration, vortex extraction, ultrasonic extraction, counter current extraction, percolation, repercolation, evacolation (extraction under reduced pressure), subcritical or supercritical fluid extraction, diacolation and solid / liquid extraction under continuous reflux. Percolation is even more preferred and has good upscaling properties. Following the extraction process, the crude extracts obtained may optionally be subjected to other typical steps, such as, for instance, fractionation, purification and / or decoloration.

[0107] Lactic acid bacteria (LAB) have found widespread usage as probiotics and various health- related applications, i.e. the irritable bowel syndrome (IBS). Preferably, the microorganism in the use according to the invention is a Lactobacillus species, preferably selected from the group consisting of Lactobacillus acetotolerans, Lactobacillus acidophilus, Lactobacillus agrestimuris, Lactobacillus alvei, Lactobacillus amylolyticus, Lactobacillus amylovorus, Lactobacillus animate, Lactobacillus apis, Lactobacillus bombicola, Lactobacillus brevis, Lactobacillus brevisimilis, Lactobacillus casei, Lactobacillus catenefornis, Lactobacillus colini, Lactobacillus corticis, Lactobacillus crispatus, Lactobacillus delbrueckii, Lactobacillus equicursoris, Lactobacillus faeni, Lactobacillus fermentum, Lactobacillus fornicalis, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus gigeriorum, Lactobacillus guizhouensis, Lactobacillus halophilus, Lactobacillus hamsteri, Lactobacillus helsingborgensis, Lactobacillus helveticus, Lactobacillus hominis, Lactobacillus huangpiensis, Lactobacillus iatae, Lactobacillus iners, Lactobacillus insectis, Lactobacillus intermedius, Lactobacillus intestinalis, Lactobacillus isalae, Lactobacillus japonicus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus juensis, Lactobacillus kalixensis, Lactobacillus kefiranofaciens, Lactobacillus kimbladii, Lactobacillus kitasatonis, Lactobacillus kullabergensis, Lactobacillus laiwuensis, Lactobacillus larvae, Lactobacillus leichmannii, Lactobacillus letivazi, Lactobacillus melliventris, Lactobacillus mobilis, Lactobacillus mulieris, Lactobacillus nasalidis, Lactobacillus panisapium, Lactobacillus paracasei, Lactobacillus paragasseri, Lactobacillus paraplantarum, Lactobacillus pasteurii, Lactobacillus plantarum, Lactobacillus porci, Lactobacillus psittaci, Lactobacillus rennanquilfy, Lactobacillus reuteri, Lactobacillus rizhaonensis, Lactobacillus rhamnosus, Lactobacillus rodentium, Lactobacillus rogosae, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus selangorensis, Lactobacillus sucicola, Lactobacillus taiwanensis, Lactobacillus terrae, Lactobacillus ultunensis, Lactobacillus vermiforme, Lactobacillus xujianguonis and Lactobacillus xylocopicola.

[0108] In a further preferred embodiment, the microorganism is selected from the group consisting of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus brevis, Lactobacillus paraplantarum, Lactobacillus reuteri, Lactobacillus pentosus, Lactobacillus buchneri, Lactobacillus sucicola, Lactobacillus bombi, Lactobacillus gasseri and Lactobacillus rodentium.

[0109] In yet another preferred embodiment, the microorganism is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum.

[0110] In another preferred embodiment, the use of the microorganism according to the invention, preferably the use of a Lactobacillus species, provides a piceol content of at least 100 ppm after a brief fermentation time. Preferably, the invention provides a piceol content of at least 100 ppm after 2h of incubation with a Lactobacillus species. Preferably, the Lactobacillus species is a Lactobacillus rhamnosus.

[0111] In a further preferred embodiment, the lactobacillus species is grown in a growth medium, preferably in a bacterial growth medium and more preferably in a Man-Rogosa-Sharpe (MRS) medium, or any MRS based medium for optimized fermentation conditions. The picein extract is added to the Lactobacillus fermentation. This can be done at the beginning of the fermentation, during the exponential growth phase, or at the stationary phase.

[0112] Subsequently, in a preferred embodiment, the Lactobacillus cells are grown until exponential growth phase, followed by the complete or partial replacement of the growth medium by a simplified medium, preferably without glucose, for the actual digestion of the picein extract. In a preferred variant, the cell density is adjusted in this step to an optical density (OD620) of 5 or higher.

[0113] In a further preferred embodiment, the picein from the plant material is directly extracted and fermented within the Lactobacillus fermentation medium without a physical separation of both steps.

[0114] In yet another preferred embodiment, the use of the microorganism for producing an antimicrobial composition, preferably comprising piceol, comprises the following steps:

[0115] -providing a growth medium comprising picein and at least one microorganism

[0116] -fermenting the mixture comprising picein and said microorganism thereby forming the antimicrobial composition, preferably comprising piceol

[0117] -optionally removing the cells of the microorganism from the mixture by centrifugation, filtration or cell lysis,

[0118] -and / or optionally inactivating said microorganism by a heat-treatment, preferably at a temperature in the range of 60 to 121 °C for 1 second to 120 minutes.

[0119] Advantageously, said heat-treatment conditions lead to a complete inactivation of the microorganism.

[0120] In yet a further preferred embodiment, the picein extract is a Spruce needle extract and / or a Baccharis magnellanica extract, and the Lactobacillus is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum.

[0121] In still another preferred embodiment, the natural extract is a Spruce needle extract and / or an extract of the plant Baccharis magellanica and the microorganism according to the invention is a Lactobacillus rhamnosus strain.

[0122] In a further preferred embodiment, the use comprises the step of downstream processing of the formed antimicrobial composition, preferably wherein the downstream processing comprises a method selected from the group consisting of centrifugation, crystallization, solidphase adsorption, filtration, reverse osmosis, precipitation, decantation, liquid / liquid extraction, vacuum distillation, falling film distillation, melt crystallization, drying and / or combinations thereof. In a preferred embodiment, the microorganisms are filtered out. In a further preferred embodiment, heat-inactivation, e.g. by tyndallization or pasteurization, or cell lysis is used as a means for inactivating said microorganisms. Further preferably, the antimicrobial composition is recovered from said mixture.

[0123] In a further preferred embodiment, the formed antimicrobial composition is substantially non- petrochemical. Said composition is particularly advantageous due to its environmental friendliness and its naturalness, thereby meeting increasing consumer needs and complying with ISO 16128. The composition is considered “substantially non-petrochemical”, if at least 75 %, preferably at least 80 %, further preferably at least 85 %, particularly preferably at least 90 %, even further preferably at least 95 % of its carbon, based on the total carbon content in the composition, is non-petrochemically-derived carbon. Preferably, at least 75 %, preferably at least 80 %, further preferably at least 85 %, particularly preferably at least 90 %, and even further preferably at least 95 % of the carbon of the piceol in the composition is non- petrochemically-derived carbon. Most preferably, all carbon of piceol in the composition is non- petrochemically-derived carbon.

[0124] To distinguish between a petroleum based - and a product which is from a renewable source (“non-petrochemical”), the skilled person knows various techniques, including the evaluation of carbon- 12 / carbon- 13 and / or hydrogen-1 / hydrogen-2 ratios and radiocarbon dating. For instance, the ASTM D6866-16 method ( ) is described as a standard method for determining the biobased content of solid, liquid, and gaseous samples. Such methods comprise or consist of radiocarbon dating. Being an unstable isotope of carbon known as14C, radiocarbon emits beta particles until finally decaying to the more stable14N. Since crude oil derived products originate from carbon sources dating back millions of years, radiocarbon has decayed. Considering that the amount of radiocarbon in “sustainable” current plant sources is known, it can hence be calculated how much of the total organic carbon is petrochemically derived.

[0125] The above liquid or solid antimicrobial composition can be employed in cosmetic and / or dermatological and / or pharmaceutical products for skin and hair care and cleansing in an an amount of 0.001 to 10 wt.%, preferably 0.01 to 5 wt.%, more preferably 0.1 to 2 wt.%.

[0126] In yet another preferred embodiment, the growth medium for producing an antimicrobial composition does not comprise Escherichia coli (E. coli). Preferably, the fermentation of the plant extract is not performed by E. coli. Further preferably, the growth medium for producing antimicrobial composition does not comprise a genetically modified organism (GMO). In still another preferred embodiment, the invention relates to the use of a microorganism, preferably a Lactobacillus species, for producing an antimicrobial composition from picein.

[0127] In yet another aspect, the invention relates to the use of shikimic acid for increasing the antimicrobial efficacy of piceol and / or of piceol for increasing the antimicrobial efficacy of shikimic acid.

[0128] The preferred embodiments and features described above apply accordingly to the use according to the invention.

[0129] The embodiments described herein can be arbitrarily combined, provided that it makes technical sense.

[0130] In the present application "At least one” refers to 1 or more, for example 2, 3, 4, 5, 6, 7, 8, 9 or more. The expression "and / or" means that a linkage exists, or an alternative is provided. For instance, “A and / or B” denotes “A or B” or both, i.e. , “A and B”.

[0131] Brief description of the figures:

[0132] Figure 1

[0133] The figure shows the results of the fermentation of a natural plant extract by a microorganism. Briefly, it can be observed that the addition of a Lactobacillus strain to both a 1 % and a 2 % natural spruce needle extract provided a significant increase in piceol concentration. In turn, picein was found to be significantly decreased compared to the concentration in the natural plant extract.

[0134] Figure 2

[0135] The figure demonstrates the conversion of picein to piceol using several Lactobacillus species. Briefly, it can be observed that Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum, and Lactobacillus plantarum convert picein to piceol. Further, the figure shows that the species differ in the speed of conversion, with L. rhamnosus having the highest conversion after 2 hours.

[0136] In the following, the invention and further advantages resulting from it are explained by means of the examples. Example 1 : Extraction of fresh spruce needles

[0137] Spruce needles and stems from small branches of Picea abies were collected and extracted using water-based extraction. The mixture was stirred, followed by cooling to room temperature. The extracted plant material was separated from the extraction solution by filtration and the extracting solvent was removed by lyophilization. Subsequently, the obtained dry extract was weighed to determine the extraction yields. Picein, piceol and shikimic acid content were quantified by HPLC.

[0138] Briefly, HPLC measurements were carried out on an Agilent Technologies HPLC-1260 system (binary pump, autosampler, thermostatted column compartment and DAD detector) using an ODS-AQ column (YMC) (150 x 3mm ID with 10 x 3mm pre column, 5 pm). Analytes were separated by gradient elution as follows: t0=10% B to t15=30% B to t20=100% B, posttime 8 min. 10% B; flow rate 0,6 ml / min; 40°C; injection volume 5 pl using buffer A (0.1% formic acid) and buffer B (0.09% formic acid in acetonitril) as mobile phase. The UV detection occurred at 280 nm. Data analysis was done using Thermo Scientific Chromeleon software (version 7.2.10 ES).

[0139] The extraction time and temperature have an impact on the amount and ratio of extracted picein and piceol. Water extraction between 30 and 100 °C is particularly suitable. Accordingly, even an addition of the spruce needles to the fermentation broth could be realized.

[0140] Example 2: Extraction of dried spruce needles

[0141] Dried spruce needles were extracted using water-based extraction for 2 hours. The mixture was stirred, followed by cooling to room temperature. The extracted plant material was separated from the extraction solution by filtration and the extracting solvent removed by lyophilization. Subsequently, the obtained dry extract was weighed to determine the extraction yields. Picein, piceol, quinic acid and shikimic acid content in the dry extract were quantified by HPLC as described above.

[0142] Example 3: Extraction of Baccharis maqellanica

[0143] Spruce needles and stems from small branches of the plant Baccharis magellanica were collected and extracted using water-based extraction. The mixture was stirred, followed by cooling to room temperature. The extracted plant material was separated from the extraction solution by filtration and the extracting solvent removed by lyophilization. The dry extract obtained was weighed to determine the extraction yield. Picein, piceol and shikimic acid content were quantified by HPLC as described above.

[0144] Example 4: In-vitro production of Piceol by fermentation using a Lactobacillus strain

[0145] To investigate the picein conversion to piceol by a Lactobacilli strain, picein was fermented exemplarily with a Lactobacillus rhamnosus strain. The bacteria were grown in Man-Rogosa- Sharpe (MRS) medium until exponential growth phase, washed and adjusted to an optical density (OD620) of 5 in a simplified medium without glucose. Said medium can be prepared for instance according to Wegkamp et al. (Wegkamp, A.; Teusink, B.; Vos, W. M. de; Smid, E. J. (2010): Development of a minimal growth medium for Lactobacillus plantarum. In Letters in Applied Microbiology 50 (1), pp. 57-64. DOI: 10.1111 / j.1472-765X.2009. 02752.x).

[0146] The digestion was performed in a total volume of 1.5 ml in closed tubes without any agitation for 3 h at 37°C.

[0147] Surprisingly, as measured by HPLC analysis, an amount of 1716 ppm piceol was produced from 5000 ppm picein. Picein content, in turn, was found to be decreased from 4220 ppm to 882 ppm.

[0148] Example 5: In-vitro production of Piceol from different plant extracts using a Lactobacillus bacterium

[0149] Lactobacillus rhamnosus was grown in MRS medium until exponential growth phase, washed and adjusted to an optical density (OD620) of 5 in a simplified medium without glucose for the actual digestion of the plant extract. Two spruce needle extracts (Picea abies, extract 1 and extract 2, see experiment 1 for details), were added and incubated at 37°C for 3h. Afterwards, picein and piceol content was determined using HPLC as described above.

[0150] Control fermentations containing the plant extract but without addition of any Lactobacillus bacteria were performed in parallel. Measuring the control 1% spruce needle extract 1 , the piceol content was found to amount to 90 ppm. In the control 2% spruce needle extract 1, a piceol content of 176 ppm was detected. Surprisingly, after fermentation using a Lactobacillus rhamnosus strain, the piceol content was found to be increased by 198 % to 268 ppm (using the 1% spruce needle extract 1). In the 2% spruce needle extract 1, piceol content was increased by 186 % to 504 ppm. Using extract 2, surprisingly, the piceol content after fermentation was even found to be increased by 614 %, i.e. to 200 ppm.

[0151] Picein content was found to be strongly decreased after the fermentation. In the 1% extract, no picein was detected after said fermentation using the Lactobacillus. Moreover, 12 ppm picein were measured in the 2% extract 1. The 2% extract 2 showed 14 ppm picein.

[0152] The data is visualized in Figure 1. Example 6: In-vitro production of Piceol by fermentation using various Lactobacillus species

[0153] Various Lactobacillus strains were analysed in terms of their ability to convert picein into piceol.

[0154] Overnight cultures were prepared as follows: 50 ml MRS medium was inoculated with 100 pl glycerin bacterial culture stock (-20 °C, OD620 1) in 100 ml Erlenmeyer flasks and incubated overnight (37°C and 80rpm). OD620nm-measurrnents were conducted. Based on the ODe20nm50 ml MRS medium was inoculated with OD620nm=0.1. Cultures were incubated for 4h (37°C and 80U / min) and subsequently transferred into 50 ml tubes and centrifuged at 4000 rpm for 4 min.

[0155] The pellet was resolved in a simplified medium without glucose (ODe20nm =10).

[0156] Said medium can be prepared for instance according to Wegkamp et al. (Wegkamp et al. 2010).

[0157] In addition, 34.8 mg Picein was dissolved in 6.942 ml of a simplified medium without glucose medium (0.5% stock solution).

[0158] 1.0 ml of the test substance solution (0.5%) was mixed with 1.0 ml of the bacterial suspension (OD620nm =10) or a simplified medium without glucose (negative control: no activity expected). The samples were incubated at 37°C and shaken at 80rpm. After 2 hours and 24 hours 200 pL of the test solution was taken, diluted with 800 pL of a simplified medium without glucose and filtered (sterile filter). The samples were analyzed by HPLC and the quantification was conducted using standards with defined concentration.

[0159] This example demonstrates that several Lactobacillus species are able to convert picein to piceol. Further, it shows that they can differ in the speed of conversion, with L. rhamnosus having the highest conversion after 2 hours. These differences do not limit the general suitability of these strains as further differences in their activity can be expected under optimized fermentation conditions, e.g. growth status, nutrients (including source, quantity and quality of the nutrients such as , C- and / or N-source), presence of oxygen, bioreactor setup (e.g., mixing device), fermentation parameters (e.g., pH, temperature, foam concentration).

[0160] The data is visualized in Figure 2.

[0161] Example 7: Antimicrobial effect of Picein and Piceol The following preservation efficacy tests (PET) were performed to compare the antimicrobial efficacy of selected compounds in a basic cosmetic formulation (emulsion), prepared as described below.

[0162] The production method comprises heating phases A and B (without Carbopol ETD 2050) separately to approx. 80 °C. Subsequently, Carbopol ETD 2050 is added to phase A, followed by homogenization. Next, phase B is added to phase A and emulsified (Ultra Turrax Stirrer, 2 min, 4000 rpm / min). The mixture is allowed to cool using a reduced stirring speed. The pH value is adjusted with Phase C. Finally, phase D is added and stirred until a homogeneous emulsion is achieved.

[0163] The test compounds were incorporated at 1%.

[0164] In the PETs the emulsions were inoculated with a defined amount of Candida albicans and Aspergillus niger. The antimicrobial load in the formulation was determined after defined time intervals.

[0165] The table above shows the antimicrobial effect of piceol against the yeast Candida albicans and the fungi Aspergillus niger over the course of 28 days. Picein decreased the concentration of said fungi to a much lesser degree. Similar differences between picein and piceol can also be found for bacteria (data not shown).

[0166] Accordingly, it can be expected that the fermented plant extract has an improved preservative efficacy compared to the untreated control extract. Example 8: Testing of Minimum Inhibitory Concentration (MIC)

[0167] The antimicrobial properties were evaluated based on growth inhibition determined using Minimum Inhibitory Concentration (MIC) tests. MIC tests were carried out in 96 well plate format using optical density (OD) as metric for microbial growth. Different concentrations, usually in the form of serial dilutions, of a given test material were evaluated with respect to the resulting microbial growth (via OD determination) and compared to positive and negative controls.

[0168] Microorganisms were cultivated under adjusted conditions based on information of the German Collection of Microorganisms and Cell Cultures (DSMZ) and stored in 10% or 50% glycerol prior to use. Afterwards, the following method was applied: Conventional growth medium (according to microorganism) was added to each well of a microplate. Subsequently, each well received one of the test materials (the test material is dissolved in a solvent e.g., in ethanol, DMSO), positive controls (water), solvent controls (e.g., ethanol, DMSO), and negative controls (benchmarks according to organism). Different concentrations of each test material were assessed. Finally, the microorganism was added in a concentration of 1x106 to 6x106 CFU / mL. Incubation of the microplates was carried out at appropriate growth conditions.

[0169] According to the resulting growth, the respective test material in the used concentration was labelled either as inhibiting or not. The lowest concentration of a test material for which a complete growth inhibition was observed was defined as the MIC value. Experiments were performed at least twice.

[0170] Potential synergistic activity of test materials was judged on the basis of the determined MIC values for each individual test compound and the respective combination of test compounds, according to the method described above. The synergy index (SI) according to Kull served as a quantitative metric. The SI is defined as follows:

[0171] To calculate the synergy index with MIC values, the Kull equation was used:

[0172] SI = (M ICmixture X PA) I MICA + (M ICmixture X PB) I MICB wherein

[0173] SI is the Synergy Index according to KUIIMICA is the MIC value for

[0174] Substance A MICB is the MIC value for Substance B

[0175] MICmixture is the MIC value for the mixture of substances A and B

[0176] PA is the proportion of the substance A in the mixture

[0177] PB is the proportion of the substance B in the mixture Synergy testing

[0178] The results of the synergy testing with respect to antimicrobial activity exemplary shown against Staphylococcus aureus along with the calculated Synergy Indices are summarized in example 9. Different weight ratios were tested. The synergism was observed for different ratios of piceol and shikimic acid. Example 9: Synergistic effect of Piceol and Shikimic acid on S. aureus

[0179] The table above shows the synergistic, i.e., more than additive antimicrobial effect of piceol and shikimic acid, exemplary on S. aureus. When used in combination, the substances can be used at a lower concentration than when used individually.

Claims

1. Claims1. Method for producing an antimicrobial composition, preferably comprising piceol, comprising the following steps:-providing a growth medium comprising picein and at least one microorganism-fermenting the mixture comprising picein and said microorganism thereby forming the antimicrobial composition, preferably comprising piceol-optionally removing the cells of the microorganism from the mixture-and / or optionally inactivating said microorganism.

2. Method according to claim 1 , in which the formed antimicrobial composition comprises a piceol content of at least 100 ppm, preferably 200 ppm, further preferably of at least 300 ppm, more preferably of at least 500 ppm, even more preferably of at least 1000 ppm, and most preferably of at least 3000 ppm piceol.

3. Method according to any of the claims above, in which the picein is a picein extract, preferably extracted from a plant selected from the group consisting of Spruce, Salix, Phagnalon Rupestre, Rhodiola Rosea, Poacynum hendersonii, Baccharis magellanica or Vauquelinia.

4. Method according to any of the claims above, in which the microorganism is a Lactobacillus species.

5. Method according to any of the claims above, in which the picein extract is a Spruce needle extract and / or a Baccharis magnellanica extract, and the Lactobacillus is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paraplantarum and Lactobacillus plantarum.

6. Antimicrobial composition, preferably comprising Piceol, obtained or obtainable by a method comprising or consisting of the steps as defined in any of the claims 1 to 5.

7. Antimicrobial composition comprising or consisting of-piceol, preferably in an excess amount compared to picein-further plant-derived substances-and optionally picein.

8. Antimicrobial composition according to claim 7, wherein the further plant-derived substances are selected from the group consisting of organic acids such as shikimic acid, quinic acid, citric acid, malic acid, succinic acid, threonic acid and lactic acid.

9. Antimicrobial composition according to claim 7 or 8, wherein the antimicrobial composition comprises or consists of piceol, preferably in an excess amount compared to picein, shikimic acid, and optionally picein.

10. Antimicrobial composition according to any of the claims 6 to 9, wherein the antimicrobial composition additionally comprises at least one microorganism, preferably a Lactobacillus species.11 . Antimicobial composition according to any of the claims 6 to 10, in which the antimicrobial composition comprises a piceol content of at least 100 ppm, preferably 200 ppm, further preferably of at least 300 ppm, more preferably of at least 500 ppm, even more preferably of at least 1000 ppm, and most preferably of at least 3000 ppm piceol.

12. Cosmetic, personal care, household, home care, pet care or pharmaceutical product comprising the antimicrobial composition according to any of previous claims.

13. Cosmetic, personal care, household, home care, pet care or pharmaceutical product according to claim 12 comprising the antimicrobial composition according to any of theprevious claims in an amount of 0.001 to 10 wt.%, preferably 0.01 to 5 wt.%, and more preferably 0.1 to 2 wt.%.

14. Use of a microorganism, preferably of a lactobacillus species, for producing an antimicrobial composition comprising piceol from a composition, preferably a plantextract, comprising picein.

15. Use of shikimic acid for increasing the antimicrobial efficacy of piceol and / or of piceol for increasing the antimicrobial efficacy of shikimic acid.