Fermented chamomile drink

Abstract

The present invention relates to a beverage composition comprising fermented chamomile or a fermented chamomile extract. A further aspect of the invention is a method for preparing the beverage composition.

Description

[Technical field]

[0001] The present invention relates to a beverage composition comprising fermented chamomile or a fermented chamomile extract. A further aspect of the invention is a method for preparing the beverage composition. [Background technology]

[0002] Chamomile tea is an herbal tea made from dried chamomile flowers and hot water. Chamomile tea has long been consumed for its taste and as a traditional folk remedy for a wide range of health problems, including gastrointestinal disorders. In ancient Egypt, chamomile tea was prescribed as a cold remedy.

[0003] Chamomile tea has an earthy flavor that is not always appreciated by consumers. Many people desire novel flavors and sweetness in their beverages, but do not like to consume flavorings, especially beverages with added synthetic flavors or sugary fruit juices. Therefore, there is commercial value in providing a chamomile beverage that does not have a strong earthy note.

[0004] Any reference to a prior art document in this specification should not be considered as an admission that such prior art is well known or forms part of the common general understanding in the art. As used herein, the words "comprises," "comprising," and similar words should not be construed in an exclusive or exhaustive sense. In other words, they are intended to mean "including, but not limited to." Summary of the Invention

[0005] The object of the present invention is to improve the state of the art and to provide a novel product which overcomes at least to some extent the above-mentioned inconveniences, or at least to provide a useful alternative. This object is achieved by the subject matter of the independent claims. The dependent claims further develop the idea of ​​the present invention.

[0006] Thus, in a first aspect, the present invention provides a beverage composition comprising fermented chamomile or a fermented chamomile extract, wherein the weight ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal is greater than 2000.

[0007] A second aspect of the invention provides a container for use in a beverage preparation machine, the container containing a beverage composition of the invention. In a third aspect, the present invention provides a method for preparing a beverage composition, the method comprising: a. soaking chamomile in water to produce an aqueous chamomile extract and soaked chamomile; b. fermenting the aqueous chamomile extract in the presence or absence of soaked chamomile; Fermentation is carried out by yeasts including Pichia yeast.

[0008] A further aspect of the present invention provides the use of chamomile fermented with Pichia kluyveri NYSC5485 (CNCM I-5525) for the preparation of a beverage.

[0009] Chamomile flowers can be made into beverage infusions by steeping the dried flowers in hot water. These beverages have an earthy note with a slight sweetness. The inventors have surprisingly found that through fermentation with Pichia yeast, the ethanol content of the chamomile beverage remains low, and the flavor generated by the fermentation covers the earthy note, enhancing the sweetness without introducing off-notes. In particular, the "green" and "woody" notes that are not well-received by some consumers in chamomile beverages are reduced. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Thus, the present invention relates in part to beverage compositions comprising (e.g. consisting of) fermented chamomile or a fermented chamomile extract having a weight ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal of greater than 2000, such as greater than 4000, for example greater than 8000, such as greater than 10000, for example greater than 12000, such as greater than 14000, for example greater than 16000, such as greater than 18000, for example greater than 20000, or even such as greater than 22000. The aldehyde 3-methylbutanal contributes a malty, solvent-like note.

[0011] The ratio can be calculated from the weight concentrations in ppm as follows:

[0012]

number

[0013] During the fermentation of sugars by Pichia, higher esters such as 3-methylbutyl acetate are produced initially, but as the fermentation progresses, higher alcohols such as 3-methylbutanol are produced. A high ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal corresponds to a pleasant aroma balance in which the malty, beer and solvent notes in the fermented beverage are reduced and the earthy notes are masked. The "tea" note, which some consumers do not appreciate, is also masked by the high ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal.

[0014] In the context of the present invention, the term chamomile refers to the flower of a daisy-like plant of the Asteraceae family, which is used to make the herbal tea "chamomile tea." Chamomile may be German chamomile (Matricaria recutita) or Roman chamomile (Anthemis nobilis). Preferably, chamomile is German chamomile. The term chamomile is sometimes written as chamomile. Chamomile contains polyphenols such as apigenin, quercetin, patuletin, and luteolin.

[0015] In the context of the present invention, the term "fermentation" refers to a process in which microbial activity brings about a change (typically a desired change) in a food or beverage. Fermentation can be by yeast and / or bacteria. Fermentation can be anaerobic or aerobic. Fermentation is one of the oldest techniques used for food preservation and enrichment.

[0016] The term "fermented chamomile" refers to chamomile that has been subjected to a process in which the activity of microorganisms such as yeasts results in a chemical change, typically a desired change, in the organic components of the chamomile. The fermented chamomile may be fermented in an aqueous medium, for example by yeast. The fermented chamomile may be dried chamomile that has been added to an aqueous fermentation medium, fermented, and then separated from the aqueous fermentation medium and dried. The fermented chamomile may also have been fermented in a "solid-state" fermentation, for example by adding a yeast starter culture to the chamomile, for example fresh chamomile flowers.

[0017] The term "chamomile extract" refers to a material extracted from chamomile, for example an aqueous extract of chamomile. The term "fermented chamomile extract" refers to a chamomile extract after fermentation of the chamomile extract, for example an aqueous chamomile extract after fermentation of the aqueous chamomile extract with yeast or the like. The fermented chamomile extract may be a dried fermented chamomile extract.

[0018] In one embodiment, the beverage composition (e.g., beverage) has a weight ratio of the sum of 2-methylbutyl acetate and 2-methylbutanol to 2-methylbutanal greater than 2000, such as greater than 4000, such as greater than 6000, such as greater than 7000, or even such as greater than 8000.

[0019] The ratio can be calculated from the weight concentrations in ppm as follows:

[0020]

number

[0021] In one embodiment, the beverage composition (e.g., beverage) has a weight ratio of the sum of isobutyl acetate and isobutanol to isobutanal greater than 400, such as greater than 500, such as greater than 600, such as greater than 700, such as greater than 800, such as greater than 900, or even greater than 950. Isobutyl acetate (IUPAC name 2-methylpropylethanoate) is the acetate ester of isobutanol. The alcohol isobutanol has the IUPAC name 2-methyl-propan-1-ol. The aldehyde isobutanal, sometimes known as 2-methylpropanal, contributes to the malty note.

[0022] The ratio can be calculated from the weight concentrations in ppm as follows:

[0023]

number

[0024] During fermentation of sugars by Pichia, higher esters such as isobutyl acetate are produced initially, but as fermentation progresses, higher alcohols such as isobutanol are produced. A high ratio of the sum of isobutyl acetate and isobutanol to isobutanal corresponds to a pleasant aroma balance and less malty beer notes in the fermented beverage.

[0025] In one embodiment, the beverage composition (e.g., beverage) has a weight ratio of the sum of 3-methylbutyl acetate, 2-methylbutyl acetate, isobutyl acetate, 2-phenylethyl acetate, and 2-phenylethanol to 2,3-butanedione greater than 60, such as greater than 200, such as greater than 400, such as greater than 500, such as greater than 600, such as greater than 700, or even such as greater than 800. This ratio can be calculated from the weight concentrations in ppm as follows:

[0026]

number

[0027] The compound 2,3-butanedione is also known as diacetyl. 2,3-butanedione provides a buttery or fatty note. A high ratio of the sum of 3-methylbutyl acetate, 2-methylbutyl acetate, isobutyl acetate, 2-phenylethyl acetate, and 2-phenylethanol to 2,3-butanedione corresponds to enhanced fruity and floral notes without the buttery and fatty notes that may be perceived as off-flavors in fermentation.

[0028] In one embodiment, the beverage composition comprises less than 10 g / kg acetic acid on a dry basis, such as less than 1 g / kg acetic acid on a dry basis, further such as less than 0.5 g / kg acetic acid on a dry basis. The acid "bite" in the flavor profile of beverages such as kombucha or kefir is not always desired by consumers seeking a milder flavor profile.

[0029] Many consumers are looking for complex, refreshing beverages as an alternative to alcoholic beverages. In one embodiment, the beverage composition has an ethanol content of less than 1.2% by weight, such as less than 0.5% by weight, such as less than 0.2% by weight, further such as less than 0.05% by weight.

[0030] The beverage composition may be selected from the group consisting of a ready-to-drink beverage, a beverage liquid concentrate, a soluble beverage powder, a dried aromatic botanical material, and combinations thereof.

[0031] The ready-to-drink beverage may be a fermented chamomile infusion or water flavored with fermented chamomile. The ready-to-drink beverage may further comprise other ingredients such as flavourings or stabilizers. The ready-to-drink beverage may be a carbonated beverage. The ready-to-drink beverage may further comprise a fruit or fruit juice such as strawberry, orange, lemon, lime, peach, pomegranate, watermelon, blackberry or cranberry. The ready-to-drink beverage may further comprise an herb such as mint, verbena or basil. The ready-to-drink beverage may further comprise a flower extract such as hibiscus or rose. The ready-to-drink beverage may include added vitamins and minerals. The ready-to-drink beverage may include replicating or non-replicating beneficial microorganisms. The beneficial microorganism may be, for example, a probiotic.

[0032] Beverage preparation devices (e.g., beverage preparation machines) containing extractable portioned ingredients provide a convenient way to prepare beverages. Such portioned ingredients are generally packaged in containers, such as configured as pods, pads, sachets, pouches, or capsules. One aspect of the present invention provides a container for use in a beverage preparation device, containing a beverage composition of the present invention. The container is for preparing a beverage when inserted into the beverage preparation device. The container may be, for example, a beverage capsule, among other configurations. In one embodiment, the container contains the beverage composition of the present invention. For example, the container may contain dried fermented chamomile or dried fermented chamomile extract. The container may contain dried fermented chamomile and / or dried fermented chamomile extract, which may be combined with other dry ingredients, such as tea, herbal tea, dried fruit, soluble coffee, or roasted and ground coffee.

[0033] One aspect of the invention provides a method for preparing a beverage composition, the method comprising: a. soaking chamomile in water to produce an aqueous chamomile extract and soaked chamomile; b. fermenting the aqueous chamomile extract in the presence or absence of soaked chamomile; Fermentation is carried out by yeasts including Pichia yeast.

[0034] The yeast may be predominantly Pichia yeast, e.g., greater than 50% of the microbial colony forming units present during the fermentation may be Pichia. For example, greater than 50% of the yeast colony forming units may be Pichia, e.g., greater than 60%, 70%, 80%, 90% of the yeast colony forming units may be Pichia. For example, essentially all yeast colony forming units present during the fermentation may be Pichia, and as a further example, essentially all microbial colony forming units present during the fermentation may be Pichia.

[0035] The beverage composition prepared by the method of the present invention may be a chamomile beverage, for example a beverage containing fermented chamomile or fermented chamomile extract.The beverage composition prepared by the method of the present invention may have an ethanol content of less than 1.2% by weight, for example less than 0.5% by weight, for example less than 0.2% by weight, and further for example less than 0.05% by weight.In one embodiment, the beverage composition prepared by the method of the present invention is a beverage composition of the present invention.

[0036] The term soaking refers to steeping the material in a liquid. The chamomile may be soaked in water for at least 2 minutes, such as at least 3 minutes, such as at least 4 minutes, such as at least 5 minutes, such as at least 10 minutes, such as at least 15 minutes, such as at least 20 minutes, such as at least 30 minutes, such as at least 60 minutes, and further such as at least 120 minutes. The components of the chamomile are extracted into the water during the soaking process. The soaking may be carried out at a temperature between 4°C and 98°C, such as between 20°C and 95°C, such as between 60°C and 95°C. Chamomile contains some fermentable sugars, but further fermentable sugars, such as sucrose or glucose, may be added to the chamomile aqueous extract. The fermentable sugars may be fructose or glucose. The fermentable sugars may be in the form of honey. For example, fermentable sugars may be added to the chamomile aqueous extract at a level of 2-10% by weight, such as 3-7% by weight. The chamomile may be dried and / or crushed prior to steeping.

[0037] If fermentation is performed in the absence of soaked chamomile, the chamomile can be soaked in water, for example using a continuous countercurrent extraction system. A further option is to soak the chamomile in water using a batch process, and the soaked chamomile is separated from the chamomile aqueous extract using a filtration system. The ratio of chamomile to water on a dry basis during soaking can be 1:1 to 1:100 by weight, for example 1:2 to 1:95 by weight, for example 1:10 to 1:90 by weight, for example 1:30 to 1:70 by weight, and further for example 1.3 to 1:6.

[0038] After soaking, the temperature of the aqueous chamomile extract is adjusted to a temperature suitable for yeast growth, for example 20°C to 37°C, for example 25°C to 35°C, for example 28°C to 32°C. The temperature can be adjusted, for example, using a laminar flow heat exchanger. The yeast is added to the aqueous chamomile extract to initiate fermentation. The fermentation of the aqueous chamomile extract may be carried out at a temperature of 25°C to 35°C, for example 28°C to 32°C. The fermentation of the aqueous chamomile extract may be carried out for at least 2 hours, for example at least 6 hours, for example at least 12 hours, for example at least 24 hours, further for example at least 48 hours.

[0039] The fermentation may be carried out at a pH of from 3 to 8, for example from 4 to 6.

[0040] Fermentation may be carried out in the presence of a nitrogen source such as yeast extract or yeast peptone.Fermentation may be carried out in the presence of amino acids such as valine, leucine, isoleucine and / or phenylalanine, added, for example, at a level of 0.05% to 0.1% by weight of the aqueous chamomile extract.

[0041] The fermentation may be carried out under anaerobic, microaerobic or aerobic conditions. For example, the oxygen partial pressure in the fermenter may be 0-30%, such as 1-20%, further such as 2-10%.

[0042] Preferably the chamomile has not been subjected to any chemical treatment, such as hydrolysis, prior to fermentation.

[0043] Fermentation should be continued for at least 10 3 CFU / g, e.g. at least 10 4 CFU / g, e.g. at least 10 5 CFU / g, e.g. at least 10 6 CFU / g, further e.g. at least 10 7 This can be done by adding one or more yeasts at an initial mixed yeast level of CFU / g.

[0044] The fermentation may be carried out in the absence of acetic bacteria to avoid the development of an acid "hot" flavor profile, as can be obtained with kefir fermentation. In one embodiment, the fermentation is carried out in the absence of lactic acid bacteria.

[0045] In one embodiment, fermentation is performed in the absence of Saccharomyces subsp. For example, fermentation can be performed in the absence of Saccharomyces cerevisiae. S. cerevisiae is used in winemaking and beer brewing. In addition to producing alcohol, which may be undesirable, S. cerevisiae produces "fermented," wine-like, yeasty flavor notes rather than fresh, fruity notes.

[0046] In one embodiment, the fermentation is carried out in the absence of yeasts selected from the group consisting of Zygosaccharomyces bailii, Schizosaccharomyces pombe, Torulaspora delbreukii, Rhodotorula mucilaginosa, Brettanomyces bruxellensis and Candida stellate, which are commonly present in kombucha "tea-fungal" cultures.

[0047] The inventors have found that fermentation of chamomile with Pichia kluyveri gives particularly good results: beverage compositions containing chamomile or chamomile extracts fermented with Pichia kluyveri have a pleasant fruity flavor with fewer grassy notes than unfermented beverages, and have an enhanced sweetness perception.

[0048] Pichia kluyveri occurs naturally throughout the world, for example in olives, grapes and coffee. In one embodiment, the Pichia yeast is predominantly Pichia kluyveri, for example, more than 50% of the microbial colony forming units present during the fermentation may be Pichia kluyveri. For example, more than 50% of the yeast colony forming units may be Pichia kluyveri, for example, more than 60%, 70%, 80%, 90% of the yeast colony forming units may be Pichia kluyveri. For example, essentially all yeast colony forming units present during the fermentation may be Pichia kluyveri, and as a further example, essentially all microbial colony forming units present during the fermentation may be Pichia kluyveri.

[0049] Pichia kluyveri NCYC246 (alternatively designated CBS188) is the type strain of Pichia kluyveri. Type strains are publicly available, for example from the National Collection of Yeast Cultures, Quadram Institute Bioscience (Norwich, UK).

[0050] In one embodiment, the Pichia yeast is Pichia kluyveri NCYC246. For example, greater than 50% of the yeast colony forming units present during the fermentation can be Pichia kluyveri NCYC246. For example, greater than 50% of the microbial colony forming units present during the fermentation can be Pichia kluyveri NCYC246. For example, essentially all of the yeast colony forming units present during the fermentation can be Pichia kluyveri NCYC246. As a further example, essentially all of the microbial colony forming units present during the fermentation can be Pichia kluyveri NCYC246.

[0051] Pichia kluyveri NCYC2781 (alternatively designated Maize Silage Isolate 86) is publicly available from, for example, the National Collection of Yeast Cultures, Quadram Institute Bioscience (Norwich, UK).

[0052] In one embodiment, the Pichia yeast is Pichia kluyveri NCYC2781. For example, greater than 50% of the yeast colony forming units present during the fermentation can be Pichia kluyveri NCYC2781. For example, greater than 50% of the microbial colony forming units present during the fermentation can be Pichia kluyveri NCYC2781. For example, essentially all of the yeast colony forming units present during the fermentation can be Pichia kluyveri NCYC2781. As a further example, essentially all of the microbial colony forming units present during the fermentation can be Pichia kluyveri NCYC2781.

[0053] Pichia kluyveri is widely distributed in nature and is present in fruits such as coffee cherries and olives. Pichia kluyveri NYSC5485, collected and isolated from a coffee fermentation plant in Nicaragua, was deposited on June 23, 2020 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25rue du Docteur Roux, F-75724 PARIS Cedex15, France, and received the accession number CNCM I-5525.

[0054] In one embodiment, the yeast is Pichia kluyveri NYSC5485 (CNCM I-5525). For example, more than 50% of the yeast colony forming units present during the fermentation can be Pichia kluyveri NYSC5485 (CNCM I-5525). For example, more than 50% of the microbial colony forming units present during the fermentation can be Pichia kluyveri NYSC5485 (CNCM I-5525). For example, essentially all of the yeast colony forming units present during the fermentation can be Pichia kluyveri NYSC5485 (CNCM I-5525). As a further example, essentially all of the microbial colony forming units present during the fermentation can be Pichia kluyveri NYSC5485 (CNCM I-5525).

[0055] The inventors have found that particularly good results are obtained by fermenting chamomile with Pichia kluyveri NYSC 5485. Fermenting chamomile with Pichia kluyveri NYSC 5485 results in a higher weight ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal, which corresponds to an improved aroma balance.

[0056] Fermentation should be continued for at least 10 3 CFU / g, e.g. at least 10 4 CFU / g, e.g. at least 10 5 CFU / g, e.g. at least 10 6 CFU / g, further e.g. at least 10 7 This can be achieved by adding Pichia kluyveri, for example Pichia kluyveri 5485 (CNCM I-5525), at an initial yeast level of CFU / g.

[0057] The gene sequence of Pichia kluyveri NYSC5485 (CNCM I-5525) was analyzed using PacBio sequencing technology. DNA purification was performed using Qiagen Gentra Puregene Yeast kit. 12 DNA fragments were identified. These fragments could be individual chromosomes, i.e. 11 chromosomes and one mitochondrial DNA (SEQ ID NO: 4). The gene sequences are submitted in electronic format as SEQ ID NO: 1 to SEQ ID NO: 12.

[0058] In one embodiment of the method of the invention, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99% identity) to SEQ ID NO:1, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:2, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:3, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:5, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:6, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:7, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:8, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:9, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:10, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:11, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:12, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:13, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:14, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:15, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ These include a gene sequence having at least 99% identity to sequence number 7, a gene sequence having at least 98% identity (e.g. at least 99% identity) to SEQ ID NO:8, a gene sequence having at least 98% identity (e.g. at least 99% identity) to SEQ ID NO:9, a gene sequence having at least 99% identity to SEQ ID NO:10, a gene sequence having at least 98% identity (e.g. at least 99% identity) to SEQ ID NO:11, or a gene sequence having at least 98% identity (e.g. at least 99% identity) to SEQ ID NO:12.

[0059] In one embodiment of the method of the invention, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99% identity) to SEQ ID NO:1, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:2, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:3, a gene sequence having at least 99.5% identity to SEQ ID NO:4, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:5, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:6, identity) to SEQ ID NO:7, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:8, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:9, a gene sequence having at least 99% identity to SEQ ID NO:10, a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:11, and a gene sequence having at least 98% identity (e.g., at least 99% identity) to SEQ ID NO:12.

[0060] In one embodiment of the method of the present invention, the Pichia yeast comprises gene sequences having at least 98% (e.g., at least 99%) overall nucleotide identity to a combination of SEQ ID NOs: 1 to 12. The term "combination of SEQ ID NOs: 1 to 12" means that a sequence identity comparison is made for all individual DNA fragments as if they were concatenated into a single sequence.

[0061] In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:1. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:2. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:3. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:5. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:6. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:7. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:8. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO:9. In one embodiment, the Pichia yeast comprises a gene sequence having at least 99% identity to SEQ ID NO: 10. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO: 11. In one embodiment, the Pichia yeast comprises a gene sequence having at least 98% (e.g., at least 99%) identity to SEQ ID NO: 12.

[0062] Names referring to yeasts may change over time, and it should be understood that a yeast according to the methods of the invention may contain the genetic sequence identities described in the preceding paragraph for the Pichia yeasts of the invention, regardless of the name applied to the yeast.

[0063] One aspect of the invention provides a method of preparing a beverage composition, the method comprising: a) soaking chamomile in water to produce an aqueous chamomile extract and soaked chamomile; and b) fermenting the aqueous chamomile extract in the presence or absence of soaked chamomile, wherein the fermentation is performed by a yeast comprising a genetic sequence having at least 98% (e.g., at least 99%) overall nucleotide identity to a combination of SEQ ID NOs:1-12.

[0064] In one embodiment, the aqueous chamomile extract is separated from the soaked chamomile after fermentation. For example, the soaked chamomile can be separated from the aqueous chamomile extract using a filtration system. The aqueous chamomile extract becomes the fermented chamomile extract after fermentation.

[0065] The fermented chamomile extract may be packaged as a ready-to-drink beverage, for example packaged in bottles or cans. The fermented chamomile extract may have a concentration of 2-60% by weight total dissolved solids, for example 2-25% by weight total dissolved solids. The fermented chamomile extract may be packaged as a ready-to-drink beverage after adding additional ingredients such as flavorings and preservatives. The fermented chamomile extract may be heat treated to inactivate or kill yeast and any spoilage organisms before packaging. The fermented chamomile extract may be passed through a filter to remove yeast and spoilage organisms before packaging.

[0066] In one embodiment, the chamomile aqueous extract is concentrated after fermentation and packaged, for example, as a liquid concentrate suitable for dilution by the consumer before ingestion. The chamomile aqueous extract may be concentrated, for example, as an intermediate product for the manufacture of ready-to-drink products at different locations. The fermented chamomile extract may have a concentration of total dissolved solids of 25-60% by weight, for example, a concentration of total dissolved solids of 50-60% by weight. The dissolved solids may also be measured, for example, by passing the extract through a fine filter to remove undissolved solids, weighing the filtrate to determine the total weight, evaporating the filtrate, and then weighing the residue to obtain the weight of the dissolved solids.

[0067] The fermented chamomile aqueous extract (e.g., fermented chamomile extract) may be concentrated to produce a powder, such as a soluble beverage powder. For example, the fermented chamomile aqueous extract (fermented chamomile extract) may be dried by spray drying or freeze drying to produce a soluble beverage powder. For example, the fermented chamomile aqueous extract may be spray dried with a carrier, such as maltodextrin, to produce a soluble beverage powder. The soluble beverage powder may be packaged and sold as is, or may be used as an intermediate product, for example for the manufacture of ready-to-drink products at a different location. The soluble beverage powder may be filled into containers for use in beverage preparation equipment. The soluble beverage powder may be combined with other raw materials, for example, dry raw materials such as tea, herbal tea, dried fruit, soluble coffee or roasted and ground coffee. For example, the soluble beverage powder may be combined with other raw materials and filled into containers for use in beverage preparation equipment. The soluble beverage powder may be combined with a milk powder or a beverage creamer powder and optionally sugar to produce a beverage mix. After fermenting the aqueous chamomile extract in the presence of the soaked chamomile, the soaked chamomile becomes fermented chamomile. In one embodiment, the soaked chamomile present during the fermentation of the aqueous chamomile extract is separated from the aqueous chamomile extract after fermentation and then dried, for example using a rotary dryer. The dried soaked chamomile may be packed into containers for use in beverage preparation equipment or into permeable bags such as "tea bags". The dried soaked chamomile may be combined with other dried raw materials such as tea, herbal tea, dried fruit, soluble coffee or roasted and ground coffee. In the context of the present invention, the term tea refers to the dried leaves, leaf buds, twigs or stems of the plant Camellia sinensis. The tea may be, for example, C. sinensis var. sinensis or C. sinensis var. assamica. The term "herbal tea" refers to an infusion made from the dried flowers, fruits, leaves, seeds, or roots of a plant other than Camellia sinensis.

[0068] In one embodiment, the soaked chamomile present during the fermentation of the aqueous chamomile extract is dried together with the aqueous chamomile extract after fermentation, for example by freeze-drying or spray-drying. The dried soaked chamomile (e.g., dried fermented chamomile) together with the dried aqueous chamomile extract after fermentation (e.g., dried fermented chamomile extract) may be packed into a container for use in a beverage preparation device, or into a liquid-permeable bag such as a "tea bag". The dried soaked chamomile and the dried aqueous chamomile extract after fermentation may be combined with other dried raw materials, such as tea, herbal tea, dried fruit, soluble coffee or roasted and ground coffee.

[0069] One aspect of the present invention provides the use of chamomile fermented with Pichia kluyveri NYSC5485 (CNCM I-5525) for the production of a beverage.

[0070] The fermented chamomile according to the invention can be obtained in "solid" fermentation, for example by adding a starter culture of Pichia yeast to chamomile, for example fresh, undried chamomile flowers. The term "solid" fermentation refers to fermentation carried out on solid material. Although liquid water is present in solid fermentation, this term is used in contrast to liquid fermentation, such as may occur in an aqueous solution in a fermentation vessel.

[0071] One aspect of the invention provides a method for preparing a beverage composition, the method comprising: adding a starter culture of yeast to the chamomile, the starter culture having at least 10 4 CFU (e.g., at least 10 5 CFU, further e.g. at least 10 6 providing yeast in a concentration of 100 CFU; The process of fermenting chamomile; drying the fermented chamomile to produce fermented chamomile; extracting the fermented chamomile with water to produce a ready-to-drink beverage; or filling the fermented chamomile into a container, for example a container for preparing a beverage when inserted into a beverage preparation device; Includes.

[0072] The yeast starter culture may be added to the chamomile as a liquid or powder. The chamomile flowers may have a moisture content of 45-55% by weight when the yeast starter culture is added. The flowers may be steamed prior to fermentation to reduce the amount of environmental microorganisms before adding the yeast. Fermentation may occur at a solids content of more than 40% by weight, such as more than 45% by weight, further such as more than 50% by weight.

[0073] While fermentation continues to some extent, drying of the fermented chamomile may be started. For example, the chamomile flowers may be placed in a container such as a tank, a starter culture may be added, and then fermentation may proceed for 1-4 days, after which the fermented chamomile may be removed from the container and dried. As a further example, after the starter culture is added to the chamomile flowers, the chamomile flowers may be gradually dried, for example, on a rack or hanging tray. Once fermentation has begun, the chamomile flowers may be gradually dried as fermentation continues. For example, the fermented chamomile flowers may be dried to a moisture content of less than 20% by weight (e.g., less than 15% by weight) over a period of 1-30 days, such as 4-20 days. The drying temperature may be less than about 40° C., for example, about 25° C. to 35° C. A fan may be used to blow dry air over the material until the chamomile flowers are dry. The fermented chamomile flowers may be dried in the sun.

[0074] The yeast may be a Pichia, such as Pichia kluyveri, further such as Pichia kluyveri NYSC5485.

[0075] When inserted into the beverage preparation device, the container for beverage preparation may be in the form of a pod, pad, sachet, pouch, capsule or the like.

[0076] The ready-to-drink beverage may be filled into bottles or cans. The fermented chamomile may be combined with other extractable ingredients such as tea, herbal tea, dried fruit, soluble coffee or roasted and ground coffee before being extracted with water to produce the ready-to-drink beverage.

[0077] The fermented chamomile may be combined with tea, herbal tea, dried fruit, soluble coffee or roasted and ground coffee in a container for preparation of a beverage when inserted into a beverage preparation device.

[0078] Those skilled in the art will understand that all features of the invention disclosed herein can be freely combined. In particular, features described for the product of the invention can be combined with the method of the invention, and vice versa. Furthermore, features described for different embodiments of the invention can be combined. Where known equivalents exist for specific features, such equivalents are incorporated herein as if specifically mentioned.

[0079] Further advantages and features of the invention are apparent from the drawings and non-limiting examples. EXAMPLES

[0080] Example 1: Fermentation method for chamomile Infusions were prepared using organic chamomile (Chamomilla recutita) (Coop Naturaplan, Switzerland) as starting material. First, a hot infusion was made by adding 20 g of dry starting material in 1000 mL of water at 100° C. and steeping for 10 minutes. The infusion was then placed in an ice bath until the temperature dropped below 30° C. The infusion was then filtered through a sieve to further remove small particles. The filtrate (chamomile aqueous extract) was used as a control (Ref) or distributed equally to the various fermentation samples.

[0081] Fermented chamomile was produced by fermenting the infusion with glucose (2% m / m) and various starter cultures including Pichia kluyveri NYSC5485 (CNCM I-5525), P. kluyveri NCYC246 (type strain of Pichia kluyveri), P. kluyveri NCYC2781, commercial brewer's yeast Saccharomyces cerevisiae (SafAle S-33, LeSaffre, France), and symbiotic culture of bacteria and yeasts (SCOBY) pellicles from kombucha tea fermentation. All strains (except SCOBY) were grown in edible yeast-peptone liquid medium from glycerol vials and inoculated into the reference chamomile brew at an optical density (OD600nm) of 0.5. All inoculated infusions were stored at 30°C under microaerobic conditions for 24 h. After incubation, each fermentation exudate was filter sterilized (0.2 μm).

[0082] Example 2: Sensory analysis of chamomile fermented with different yeasts The sensory profiles of the different variations of chamomile infusions were tasted. The control had a floral and green taste with earthy notes and slight sweetness. The chamomile infusion fermented with Saccharomyces cerevisiae had a malty and beer-like character. The chamomile fermented with SCOBY had a very strong sour taste. The chamomile infusions fermented with two different Pichia yeasts had enhanced sweetness, some banana / fruity notes, and significantly reduced green and earthy notes compared to the control. No buttery or fatty off-notes were identified in the chamomile fermented with Pichia yeast. The chamomile fermented with Pichia kluyveri NYSC5485 was also found to have a cleaner aroma profile and less pronounced banana notes than the chamomile fermented with Pichia kluyveri NCYC246.

[0083] Example 3: Analysis of the flavor profile focusing on aroma compounds present in chamomile fermented with different yeasts Quantitative profiling of volatile compounds in chamomile infusions was performed by headspace / solid-phase microextraction coupled with gas chromatography and mass spectrometry (HS / SPME-GC-MS) using a gas chromatograph (Agilent Technology 8890 GC System) coupled to a mass spectrometer (Agilent Technology 7010B TQ).

[0084] A 1.0 mL sample of chamomile infusion was first incubated in a 10 mL screw-top headspace vial at 30°C for 10 min and then extracted using an SPME fiber (PDMS / DVB, Supelco) at 30°C for 10 min. All vials were placed in a tray cooled to 6°C before analysis. Volatiles were thermally desorbed from the SPME fiber at 250°C in splitless mode and separated on a capillary column (DB-WAX, Agilent). The gas chromatograph oven temperature was programmed to be first at 35°C for 5 min, then increased to 230°C at 4°C / min, and then at 230°C for 10 min. Helium was used as the carrier gas at a flow rate of 1 mL / min. Target compounds were identified by pure standards and quantified with MSD ChemStation software (Agilent). All samples were run in duplicate.

[0085] The levels of 3-methylbutyl acetate, 3-methylbutanol, 3-methylbutanal, 2-methylbutyl acetate, 2-methylbutanol, 2-methylbutanal, isobutyl acetate, 2-phenylethyl acetate, 2-phenylethanol, 2,3-butanedione, ethanol and acetic acid after 24 hours are shown in the table below in ppm.

[0086] [Table 1]

[0087] The SCOBY fermented sample had high levels of acetic acid equivalent to 200 g / kg on a dry basis.

[0088] The weight ratios A, B, C and D of the aroma components are shown in the table below.

[0089]

number

[0090] [Table 2] Samples fermented with Pichia yeasts (NYSC5485 and NCYC246) show higher Ratio A, Ratio B, Ratio C and Ratio D than the control and samples fermented with S. cerevisiae or SCOBY. P. kluyveri NYSC5485 produces significantly higher values ​​for Ratio A, Ratio B, Ratio C and Ratio D than the reference strains P. kluyveri NCYC246 or P. kluyveri NCYC2781.

[0091] The levels of the terpene myrcene present in the samples were measured. Compounds were identified by NIST library and retention index. Peak areas were integrated over the entire sample. The ratios of fermented sample peak area to non-fermented sample peak area are shown below.

[0092] All samples were measured in duplicate.

[0093] [Table 3]

[0094] Myrcene has green, woody, "hop-like" notes (which are also present in hops and cannabis). These notes are not always appreciated by consumers. After fermentation with Pichia kluyveri yeast, myrcene (related to hop-like characteristics) was significantly reduced. P. kluyveri NYSC 5485 was found to reduce myrcene in contrast to the reference strain P. kluyveri NCYC 246.

[0095] Example 4: Investigation of the effect of aroma ratio on sensory perception by spikes Solutions were prepared with different levels of 3-methylbutyl acetate, 3-methylbutanol and 3-methylbutanal added: in one set of solutions, the compounds were added to water, in the other set, the compounds were added to unfermented chamomile extract (Ref).

[0096] The samples were tasted by a four-person expert taste panel and comments were collected. The samples and comments are shown below.

[0097] [Table 4]

[0098] [Table 5]

[0099] It is important to have the right balance between esters / alcohols and aldehydes. It has been observed that in chamomile infusions, high levels of esters and low levels of aldehydes, for example ratios A above 2000, can mask the "tea" note.

[0100] Example 5: Comparison of the gene sequence of P. kluyveri NYSC5485 with that of the reference strain P. kluyveri NCYC246 The gene sequences of Pichia kluyveri NYSC5485 (CNCM I-5525) and P. kluyveri NCYC246 were analyzed using PacBio sequencing technology. DNA purification was performed using the Qiagen Gentra Puregene Yeast kit. 12 DNA fragments were identified for P. kluyveri NYSC5485 and 15 DNA fragments were identified for the type strain P. kluyveri NCYC246. The gene sequence of P. kluyveri NYSC5485 was submitted in electronic form as SEQ ID NO:1 to SEQ ID NO:12, and for P. kluyveri NCYC246 as SEQ ID NO:13 to SEQ ID NO:27. Mitochondrial DNA is SEQ ID NO:4 for P. kluyveri NYSC5485 and SEQ ID NO:20 for P. kluyveri NCYC246.

[0101] The gene sequence of Pichia kluyveri NYSC5485 (CNCM I-5525) was compared with P. kluyveri NCYC246 using the software OrthoANIu. The overall average nucleotide identity was found to be 97.91%.

[0102] The correspondence between the various sequences is shown in the table below. The multiple sequences of Pichia kluyveri NYSC5485 correspond most closely to the combination of sequences of P. kluyveri NCYC246 and vice versa.

[0103] [Table 6] The type strain P. kluyveri NCYC246 contained the gene sequences with the greatest identity to each of SEQ ID NOs: 1-12 as shown below:

[0104] SEQ ID NO:1, 97.85%; SEQ ID NO:2, 97.40%; SEQ ID NO:3, 97.86%; SEQ ID NO:4, 99.02%; SEQ ID NO:5, 98.05%; SEQ ID NO:6, 97.93%; SEQ ID NO:7, 98.12%; SEQ ID NO:8, 97.07%; SEQ ID NO:9, 97.20%; SEQ ID NO:10, 98.33%; SEQ ID NO:11, 97.526%; SEQ ID NO:12, 97.77%.

Claims

1. A beverage composition comprising fermented chamomile or a fermented chamomile extract, wherein the weight ratio of the sum of 3-methylbutyl acetate and 3-methylbutanol to 3-methylbutanal is greater than 2,000.

2. 2. The beverage composition of claim 1, wherein the weight ratio of the sum of 2-methylbutyl acetate and 2-methylbutanol to 2-methylbutanal is greater than 2,000.

3. 2. The beverage composition of claim 1, wherein the weight ratio of the sum of isobutyl acetate and isobutanol to isobutanal is greater than 400.

4. 2. The beverage composition of claim 1, wherein the weight ratio of the sum of 3-methylbutyl acetate, 2-methylbutyl acetate, isobutyl acetate, 2-phenylethyl acetate, and 2-phenylethanol to 2,3-butanedione is greater than 60.

5. A beverage composition according to any one of claims 1 to 4, having an ethanol content of less than 1.2% by weight.

6. 5. The beverage composition of any one of claims 1 to 4, wherein the beverage composition is selected from the group consisting of ready-to-drink beverages, beverage liquid concentrates, soluble beverage powders, dried aromatic plant materials, and combinations thereof.

7. A container for use in a beverage preparation machine, the container containing a beverage composition according to any one of claims 1 to 4.

8. 1. A method for preparing a beverage composition, comprising: a. soaking chamomile in water to produce an aqueous chamomile extract and soaked chamomile; b. Fermenting the aqueous chamomile extract in the presence or absence of the soaked chamomile; The method, wherein the fermentation is carried out by a yeast, including a Pichia yeast.

9. 9. The method of claim 8, wherein the Pichia yeast is Pichia kluyveri.

10. 9. The method of claim 8, wherein the Pichia yeast comprises a gene sequence having at least 98% overall nucleotide identity to a combination of SEQ ID NOs: 1 to 12.

11. The method according to any one of claims 8 to 10, wherein the fermentation is carried out in the absence of Saccharomyces cerevisiae.

12. 11. The method of any one of claims 8 to 10, comprising separating the aqueous extract of chamomile from the soaked chamomile after fermentation.

13. 11. The method according to any one of claims 8 to 10, wherein the steeped chamomile present during the fermentation of the aqueous chamomile extract is dried together with the aqueous chamomile extract after fermentation.

14. 9. The method of claim 8, wherein the beverage composition is as defined in claim 1.

15. Use of chamomile fermented with Pichia kluyveri NYSC5485 (CNCM I-5525) for producing a beverage.