FOOD PRODUCTS, ESPECIALLY BAKED PRODUCTS ENRICHED WITH GABA AND BENEFICIAL TO HEALTH

Abstract

The present invention relates to a natural extract, preferably extracted from yeast, rich in gamma-aminobutyric acid (GABA), a product comprising such an extract, and its use in preparing food products for human or animal consumption that are beneficial to health and well-being. Such foods include, in particular, baked goods such as bread.

Description

Title of the invention: FOOD PRODUCTS, IN PARTICULAR BAKERY PRODUCTS ENRICHED WITH GABA AND BENEFICIAL TO HEALTH technical field

[0001] The present invention falls within the field of agri-food and health, seeking natural solutions to improve the nutritional and medicinal qualities of ingested products. The solution proposed in this application is the provision of a natural extract rich in GABA, whose beneficial health properties are known (Pydi et al., J Biol Chem., 2014, 289(36): 25054-66; Sasaki et al., J Biol Macromol., 2014, 14(1): 43-57). Thus, by way of example, the incorporation of such an extract during a bread-making process makes it possible to obtain "healthy" bread for humans. Prior state of the art

[0002] The non-proteinogenic amino acid GABA (gamma-aminobutyric acid) is naturally present in microorganisms, plants, and animals, where this bioactive compound plays various roles. Thus, GABA is thought to be involved in mammalian behavior by regulating stress and anxiety, modulating cognitive and brain functions, promoting sleep, and improving mood (Rashmi et al., 2018, Studies in Natural Products Chemistry, 57: 413-452).

[0003] In practice, positive effects of GABA administration in the context of certain chronic diseases have been observed in clinical trials:

[0004] Inoue et al. (2003, European Journal of Clinical Nutrition, 57: 490-495) conducted a randomized, single-blind, placebo-controlled trial in 39 hypertensive patients to evaluate the effect of daily intake of 100 ml of fermented milk containing 10 to 12 mg of GABA for 12 weeks. They observed a decrease in blood pressure in these patients and concluded that daily intake of fermented milk containing GABA may be useful for controlling blood pressure levels in mildly hypertensive patients.

[0005] Okada et al. (2000, Journal of the Japanese Society for Food Science and Technology, 47(8): 596-603), in a double-blind test, identified a potential treatment for patients suffering from neurological disorders (insomnia, depression and autonomic disorders) by taking sprouted rice enriched with GABA (26.4 mg / day).

[0006] The recommended intake and toxic dose of GABA have not yet been established, but clinical studies based on the administration of a high dose of this compound (300 mg per day) revealed no serious adverse effects after four weeks of consumption (Byun et al., 2018, J Clin Neurol., 14(3): 291-295).

[0007] The human body is capable of producing its own reserve of GABA, but in small quantities, which can also be inhibited by a lack of estrogen, zinc, or vitamins, or by an excess of salicylic acid and food additives. Furthermore, the typical daily human diet is relatively low in GABA, in any case insufficient to produce relevant biological effects.

[0008] Consequently, consideration has been given to providing exogenous GABA to humans to maintain or even improve their health.

[0009] Today, most GABA-based products are in the form of dietary supplements (tablets and capsules), while only a few GABA-based foods and beverages are available on the market. For example, Kittibunchakul et al. (2021, Journal of Functional Foods, 86, 1047140) proposed a probiotic beverage based on brown rice, naturally rich in GABA, fermented using the lactic acid bacterium Lactobacillus pentosus, which also produces GABA.

[0010] However, there is an obvious need to develop new GABA-rich foods advantageously of natural origin, including foods that are consumed universally and habitually.

[0011] The object of the present invention is to provide a natural extract containing gamma-aminobutyric acid (GABA) in a complex matrix (peptides, amino acids, RNA, etc.). Thus, the integration of such an extract into an ingestible product makes it possible to provide a sufficient quantity of GABA to obtain a beneficial effect on human or animal health, in a natural context also rich in other potentially useful compounds (in terms of nutrition, health, taste, etc.). Detailed description of the invention

[0012] Thus and according to a first aspect, the present invention relates to a natural extract rich in gamma-aminobutyric acid (GABA).

[0013] In the context of the invention, "GABA-rich extract" means that the extract comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%, or even at least 11, 12, 13, 14, or even at least 15% by weight of GABA, the percentage by weight being expressed relative to the dry weight of the extract. Advantageously, the extract comprises at least 5% by weight of GABA. A content between 5 and 15% is particularly suitable for the purposes of the present invention.

[0014] In the context of the invention, a natural extract is defined as being obtained from an element of nature, in particular from plant matter or from a microorganism such as yeast or bacteria. According to one embodiment In particular, the extract according to the invention is obtained from a yeast, hereinafter referred to as "yeast extract" ("YE" for Yeast Extract).

[0015] Generally speaking, yeast extracts are products known to those skilled in the art. According to the invention, "yeast extract" (or "yeast hydrolysate" or "yeast peptone") refers to the soluble fraction obtained after thermal, mechanical (using known methods such as high-pressure homogenization, mechanical milling, mechanical lysis using glass beads, ultrasonic disintegration, repeated freeze-thaw cycles, or osmotic shock), or enzymatic lysis (using an enzyme exogenous or endogenous to the yeast)... The objective of such lysis is to release the internal macromolecules of said yeast in their native state, in particular the pool of free amino acids, including free glutamic acid. The co-products obtained correspond to the insoluble fraction called "yeast hulls" and can be used in other processes.The processes for obtaining yeast hulls and yeast extracts are known in the art (see, for example, the reference work “Yeast Technology”, 2nd edition, 1991, G. Reed and TW Nogodawithana, published by Van Nostrand Reinhold, New York, ISBN 0-442-31892-8). The insoluble fraction and / or the soluble fraction can then be dried.

[0016] Thus, a yeast extract can be in dry form, preferably as a fine water-soluble powder, in liquid form or even as a concentrated liquid, or in paste form. A yeast extract consists mainly of protein matter, preferably at least 55% protein matter.

[0017] The difficulty overcome by the present invention is to obtain a natural extract rich in GABA, without the addition of exogenous GABA. Indeed, some plants, such as broccoli or sweet potatoes, contain GABA but in very small quantities, unusable for the desired properties.

[0018] Thus, and advantageously, the natural extract used in the context of the present invention is an extract that is naturally rich in glutamic acid (or glutamate). It should be noted that, in the context of the invention, "glutamic acid" (or "glutamate") refers to the amino acid in its free form, not incorporated into a peptide or protein.

[0019] In particular, an extract is said to be "rich in glutamic acid" if it contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%, or even 11, 12, 13, 14, 15, 20, or 25% by weight of free glutamic acid, the percentage by weight being expressed relative to the dry weight of the extract. Advantageously, the extract comprises at least 10% by weight of free glutamic acid. A content between 5 and 25%, for example between 10 and 20%, is particularly suitable within the scope of the present invention. Natural extracts meeting this definition are, for example, yeast extracts, bacterial extracts or plant extracts, advantageously yeast extracts.

[0020] According to a particular embodiment, such an extract is obtained from an organism, in particular a plant or microorganism such as yeast or bacteria, capable of synthesizing or even secreting glutamic acid, advantageously in large quantities. As is known to those skilled in the art, the organism may be naturally capable of synthesizing glutamic acid in large quantities, or genetically modified for this purpose.

[0021] A yeast extract according to the invention can be obtained from any yeast, advantageously meeting the above definition. Preferably, the yeast strain used for preparing the extract according to the invention belongs to the genus Saccharomyces, Kluyveromyces, or Candida (also known as Pichia or Lindnerd). Preferably, the yeast strain used for preparing the extract belongs to the genus Saccharomyces and, more particularly, to the species Saccharomyces cerevisiae.

[0022] An example of bacteria meeting the above definition is, for example, the species Corynebacterium glutamicum.

[0023] As reported in the literature, many plants or sources of plant material meeting the above definition are known.

[0024] Obtaining GABA from glutamic acid involves the implementation of a glutamate decarboxylase (GAD) activity capable of ensuring the following conversion: L-Ghilanmter ® A GABA-r (XX

[0025] Following this step, carried out under conditions that promote the conversion of glutamic acid to GABA while ensuring limited bacterial growth, the resulting extract is low in glutamic acid, or even free of glutamic acid. For the purposes of this invention, a "low-glutamic acid extract" is defined as an extract that advantageously comprises less than 10%, or even less than 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, or even 0.2 or 0.1% by weight of glutamic acid in free form, the percentage by weight being expressed relative to the dry weight of the extract. The term "glutamic acid-free or glutamic acid-deprived extract" refers to an extract that does not contain glutamic acid in free form, corresponding to the case where all the free glutamic acid present in the extract has been converted to GABA.

[0026] In a manner known to those skilled in the art, the natural extract, advantageously the yeast extract, can be incubated in the presence of such an enzyme under conditions, in particular of temperature and pH, enabling this conversion to occur. These conditions vary depending on the origin of the GAD enzyme. For example, and in relation to the enzyme produced by Levilactobacillus brevis, these conditions are: - an acidic pH, for example of 5; and - a temperature between 30 and 35°C, for example equal to 33°C.

[0027] Alternatively, a natural extract, advantageously a GABA-rich yeast extract according to the invention, is obtained via a bacterium exhibiting suitable glutamate decarboxylase activity.

[0028] Thus, the bacterium implemented within the framework of the invention exhibits glutamate decarboxylase (GAD) activity, in other words is capable of producing a glutamate decarboxylase (GAD) enzyme. The proteins involved in this metabolic pathway, in particular the GadA and / or GadB genes encoding the enzyme and the GadC gene encoding the substrate transporter, are widely documented in the prior art, see for example the review by Yogeswara et al. (Microorganisms 2020,8(12),1923; https: / / doi.org / 10.3390 / microorganisms8121923) in connection with lactic acid bacteria.

[0029] According to a particular embodiment, a bacterium of interest carries a GadA and / or GadB gene encoding the GAD enzyme.

[0030] According to another embodiment, the bacterium further carries a GadC gene encoding a glutamate and GABA transporter.

[0031] These may be endogenous or exogenous genes, single or multicopy, chromosomally integrated or carried by a plasmid, and placed under the control of regulatory elements allowing their expression, such as a promoter. In a particular embodiment, these are endogenous genes.

[0032] It has been shown that lactic acid bacteria, particularly those of the Lactobacillaceae family, are particularly well-suited for implementation within the scope of the present invention. The bacteria listed below can be used alone or in combination.

[0033] Bacteria carrying at least one GadA and / or GadB gene are for example chosen from the group including: Lactobacillus plantarum.

[0034] Des bacterie porteuses d'au moins un gène GadA et / ou GadB et d'au moins un gène GadC sont par exemple chois dans le groupe comprendre les geneurs suivants : Levilactobacillus, Lentilactobacilus, Lactococcus, Lacticaseibacillus, Furfurilactobacillus, Leuconostoc, Lactiplantibacillus, Bifidobacterium, Pediococcus, Enterococcus et Limosilactobacillus.

[0035] For example, it may be: Levilactobacillus brevis, Lentilactobacillus buchneri, Lactococcus lactis, Lacticaseibacillus paracasei, Furfurilactobacillus rossiae, Enterococcus faecium, Leuconostoc suionicum, Lactobacillus amylovorus, Lactiplantibacillus plantarum, Bifidobacterium dentium, Bifidobacterium adolescentis, Bifidobacterium angulatum, Pediococcus pentosaceus, Enterococcus casseliflavus , Limosilactobacillus reuteri, Enterococcus gallinarum, Limosilactobacillus oris, Pediococcus acidilactici, Limosilactobacillus fermentum and Enterococcus hermanniensis.

[0036] Preferably, the bacterium, used alone or in mixture, belongs to a species chosen from the following group: Levilactobacilus brevis, Lentilactobacilus buchneri, and Lactococcus lactis, advantageously Levilactobacilus brevis.

[0037] As is known to those skilled in the art, the bacterium is cultured under conditions favorable to the growth of bacteria of the Lactobacillaceae family, for example in an MRS (Man Rogosa, Sharpe) type culture medium. The culture conditions, particularly those relating to pH, temperature, and aerobic or anaerobic conditions (partial or total), depend on the strain chosen. As is known and as described in the examples, a strain of Levilactobacillus brevis is advantageously cultured under partially anaerobic conditions, at a temperature between 30 and 35°C and at a slightly acidic pH, for example, 6.2.

[0038] Advantageously, the bacterium is further cultured under conditions promoting the expression of genes of the pathway, in particular GadA, GadBet / or GadC.

[0039] At the end of growth and for the conversion stage, the culture thus obtained can be used as is, or in dried or even freeze-dried form, provided that these treatments do not affect the enzymatic activity.

[0040] By using a bacterium equipped with the GadC-encoded transporter, whole cells can be implemented to ensure the conversion of glutamic acid from the natural extract into GABA.

[0041] Alternatively, and in particular when the bacteria do not possess such a transporter, the bacteria are subjected to lysis, then possibly centrifuged, and it is the lysate containing the enzyme of interest, possibly purified, that is used.

[0042] The extract, advantageously of yeast, is then incubated with the bacterial culture, possibly in the form of a bacterial lysate.

[0043] According to one embodiment, the free glutamic acid titration of the extract, particularly of yeast, is adjusted. In practice, the extract is advantageously used in this incubation step at a concentration of at least 100, 150, 200, 250 or even 300 g / L, for example between 150 g / L and 250 g / L.

[0044] According to one embodiment, bioconversion is ensured by bringing the extract, advantageously of yeast, as detailed above, into contact with the bacterium, namely the bacterial must or bacterial lysate, as described above.

[0045] Incubation is advantageously carried out under conditions adapted to the bacterium used or to the enzyme in the case of a bacterial lysate.

[0046] Thus, in a manner known to those skilled in the art, the incubation conditions are adjusted according to the bacterial strain and the enzyme present, to ensure optimal bioconversion activity.

[0047] The conversion step is carried out until the desired quantity of free glutamic acid is consumed. In the context of this application for the use of the final extract for health purposes, and in the absence of GABA toxicity, the conversion to GABA must be optimal, advantageously exceeding 90%. As already stated, the final extract advantageously comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1%, or even 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or even 0.1% by weight of free glutamic acid, the weight percentage being expressed relative to the dry weight of the extract. Even more advantageously, all of the glutamic acid in the extract is consumed.

[0048] According to a particular embodiment, an extract according to the invention further has the following characteristics, expressed as a percentage by weight relative to the dry weight of the extract: - a nitrogen content of 0 to 20%, for example 5 to 15%; and / or - a free amino acid content of 0 to 30%, for example 10 to 20%; and / or - a total amino acid content of 20 to 50%, for example 30 to 40%.

[0049] After the conversion has stopped, several processing schemes for the extract thus obtained can be considered:

[0050] According to a first embodiment and at least in the case of incubation with live bacteria, it is subjected to thermal inactivation, for example at a temperature of 90°C. In this case, the product obtained contains inactivated bacteria.

[0051] According to another embodiment, after this thermal inactivation step, the bacteria are removed from the extract by any technique known to those skilled in the art, for example by tangential membrane filtration (microfiltration or ultrafiltration), by centrifugation, or by a combination of these different techniques.

[0052] These steps may prove unnecessary in the case of the implementation of a bacterial lysate.

[0053] Thus and at this stage, the extract may contain inactivated bacteria or be free (or essentially devoid) of bacteria.

[0054] According to another feature, an extract according to the invention is water-soluble.

[0055] According to a particular embodiment, the extract thus obtained is dried, advantageously by spray drying, for example using a spray drying tower. In the context of this application, it has been shown that the GABA present in the extract is resistant to these heat treatments and remains stable.

[0056] In one particular embodiment, the extract according to the invention is in the form of a dry extract. Alternatively, it may be in liquid or powder form. In another particular embodiment, the extract according to the invention may be diluted in a physiologically acceptable carrier or excipient. A physiologically acceptable carrier or excipient is an aromatically neutral carrier or excipient suitable for administration in humans or animals. Examples of physiologically acceptable carriers or excipients include maltodextrins, triacetin, propylene glycol, vegetable glycerin, glycerol, soluble fibers, yeast derivatives such as yeast extracts, barks, and autolysates, or fats such as palm oil.

[0057] An extract according to the invention has health-beneficial properties of interest for any product intended for oral administration, i.e., for ingestion. One aspect of the invention therefore relates to the use of such an extract in this context.

[0058] Thus, the invention relates to a food product containing an extract as described above and the use of such an extract to obtain a food product beneficial to health, both in humans and animals.

[0059] A food product within the meaning of the present invention can be considered any substance that provides nutrition to a living being, and particularly an animal or a human being. The food product can therefore be in either solid or liquid form. By way of example, for human consumption, a food product could be a prepared meal, jam, a fermented beverage such as wine or beer, an energy drink, or a protein drink for sports. For animal feed, it could be treats, kibble, or pâté.

[0060] In practice, a food enriched with an extract according to the invention, in particular enriched with GABA and capable of influencing a wide variety of functions involved in the state of well-being and health, or the reduction of the risk of diseases, corresponds to a functional food, also called a nutraceutical or medicinal food.

[0061] Alternatively, it may be a food supplement, the extract according to the invention being the principal ingredient. By definition, a food supplement (or additive or supplement) is a foodstuff intended to supplement the normal diet and which constitutes a concentrated source of nutrients or other substances having a nutritional or physiological effect. With regard to animals, such a supplement may be added to drinking water or to feed such as, for example, cereals and / or legumes.

[0062] In the context of the invention, a prime food product is bread. Indeed, bread is a staple food consumed by 80% of global consumers. at a high frequency and the increasing demand for healthier options is a dominant trend in the bread market.

[0063] Note that the bread is obtained by a mixture of several ingredients including flour and yeast which may contain traces of GABA but in insufficient quantity for the desired effect.

[0064] Thus, the raw materials used in breadmaking, essentially flour and yeast, generally have glutamic acid and GABA levels between 0 and 50 ppm.

[0065] In the context of this application, it has been demonstrated that the integration of an extract according to the invention was compatible with breadmaking processes and made it possible to obtain a bread rich in GABA, in addition to the favorable effects of the yeast extract in particular on the volume of the bread, without degrading its olfactory qualities.

[0066] However, a number of technical prejudices explained why this solution had never been considered: Indeed, breadmaking is characterized by a series of steps likely to reduce the amount of GABA or affect its integrity, particularly during fermentation, as yeast can consume glutamate and GABA, and then during baking, as GABA participates in Maillard reactions involving reducing sugars and amino acids. By promoting these reactions, there was a fear that its addition would disrupt the bread's flavor.

[0067] According to another aspect, the present invention therefore relates to the use of a natural extract as described above in a baking or bread-making process, in particular to make a bread enriched with GABA.

[0068] For the purposes of this application, "breadmaking" or "bread production" refers to all the operations involved in transforming flour into bread. The extract according to the invention is intended to be used in any type of breadmaking, whether on a small or large scale.

[0069] More generally, an extract according to the invention can be used in the preparation of any bakery product including bread, pastries and cakes.

[0070] As is known, bread making requires flour, water, salt, and a raising agent, in particular baker's yeast and / or sourdough starter. The flour provides fermentable sugars used by the yeast and proteins, especially gluten, giving the bread dough its viscoelastic texture. The fermentative microflora, provided by the baker's yeast or sourdough starter, ferments the carbohydrates into carbon dioxide (CO2) and ethanol. The gas fills the alveoli of the gluten network and makes the dough rise, while the ethanol evaporates during baking.

[0071] Yeasts used for breadmaking, also called "baker's yeast", are living cells generally obtained from different strains of the genus Saccharomyces, in particular Saccharomyces cerevisiae, Saccharomyces chevalieri, Saccharomyces bayanus or Saccharomyces boulardii, advantageously of the species Saccharomyces cerevisiae.

[0072] They can be in several forms: yeast cream, pressed yeast, dry yeast, or frozen yeast. Fresh yeasts are characterized by a high water content compared to dry yeasts. Fresh yeasts include both yeast creams and pressed yeasts. Yeast creams, also called "liquid yeasts," are aqueous suspensions of yeast cells with a cream-like viscosity. These aqueous suspensions of live yeast cells generally have a dry matter content of at least 12% by weight, particularly from 12 to 40% by weight. A yeast cream may, for example, have a dry matter content of between 12 and 25% by weight, preferably between 14 and 22% by weight. Pressed yeasts include both compressed block yeasts and crumbled compressed yeasts.Pressed yeast in compact blocks, also called "yeast loaves," is characterized by a dry matter content between 26% and 35%. Crumbled pressed yeast has a water content between 21% and 35%. Dry yeast is characterized by a dry matter content greater than approximately 92%. Frozen yeast is characterized by a dry matter content between 74% and 80%.

[0073] Unlike a “baker’s” yeast, a natural extract, advantageously a yeast extract according to the invention, does not have fermentative power, thus not allowing fermentation to occur during breadmaking.

[0074] In the present invention, "sourdough" means a bread-making starter resulting from a mixture of flour and water in which the metabolic activity of a heterogeneous population of lactic acid bacteria and yeasts takes place, either through spontaneous fermentation (so-called "natural" sourdough) or through fermentation initiated by a starter culture, with or without refreshing. This term also encompasses industrially produced sourdoughs such as active live sourdough or starter sourdough.

[0075] Conventional breadmaking processes generally include the following steps: - Kneading: mixing the ingredients constituting the dough, called bread dough - Dividing: separating the dough (according to weight) - Shaping: forming the dough into loaves - Proofing: fermentation, generally at a temperature between 15 and 45 °C, for one to several hours - Cooking: for example at 230°C for 30 minutes.

[0076] The breadmaking processes covered by this application may be of the direct scheme type (“NO-TIME DOUGH”) or of the indirect scheme type (“SPONGE and DOUGH”).

[0077] A direct scheme involves virtually no first fermentation between intensive kneading and the division of the dough, the resulting dough pieces being fermented in a mold between 35°C and 40°C, then baked.

[0078] A Sponge and Dough type breadmaking process is described, for example, in the reference book "Bakers Handbook" by EJ Pyler, published by Sosland Publishing Co. A "SPONGE and DOUGH" scheme is a breadmaking process comprising two fermentation stages:

[0079] - a first stage, called "SPONGE", which corresponds to the fermentation of a dough comprising 50 to 70% of the total flour used, some of the water and all or part of the yeast, for several hours, generally between 3 and 6 hours, and more particularly between 3 and 4 hours; and

[0080] - a second stage, called "DOUGH", in which the SPONGE obtained after The above fermentation is combined with the rest of the flour, the rest of the water and the other ingredients of the dough, the mixture thus formed is kneaded, divided, put in a mold and fermented, then baked, this second fermentation in the mold corresponding to the proofing.

[0081] In the present invention, "dough" or "baking dough" refers to the mixture of ingredients used in preparing the final bread product. Typically, such dough comprises flour, water, salt, and a raising agent such as yeast.

[0082] Typically, yeast represents between 0.5 and 10% by weight, and preferably from 0.5 to 3% by weight relative to the total weight of the composition, namely the dough.

[0083] The extract according to the invention can be incorporated at any time during the preparation process before the cooking stage. Preferably, the natural extract, advantageously yeast extract, is added during the kneading stage, along with the ingredients previously mentioned.

[0084] According to a particular aspect, the present invention relates to a bread dough comprising an extract as defined above and the bread or bakery product obtained from said dough.

[0085] In the context of the invention, a dough may be a dough without added sugar, a slightly sweetened dough or a sweetened dough, the sugar being most often sucrose.

[0086] In the present invention, "bread" means any product obtained by baking dough made by kneading a mixture comprising at least flour, water, and salt, and leavened with a raising agent, in particular yeast. More generally, "bread product" or "product of bakery » any product resulting from breadmaking, including pastries and cakes.

[0087] According to another aspect, the present invention relates to a bread product, such as a loaf of bread, obtained from a dough comprising an extract according to the invention, in particular after fermentation and baking.

[0088] Advantageously, such a product is rich in GABA.

[0089] In the context of the invention and in connection with bread dough, bread product or more generally a food product, "GABA-rich product" means that the product includes at least 0.01, 0.015, 0.02, 0.03, 0.04, 0.05%, or even 0.1 or even 0.5% by weight of GABA, the percentage by weight being expressed in relation to the weight of the product or dough. In other words, a food product, a bakery product or a bakery dough according to the invention advantageously comprises at least 100, 150, 200, 300, 400, 500, or even 1000 or even 3000 ppm of GABA, preferably an amount between 100 ppm and 3000 ppm, even more preferably between 150 ppm and 500 ppm, for example 200 ppm.

[0090] Assuming that a man consumes about 80 g of bread per day, the daily intake of GABA via such a product could therefore be at least 1 mg, 5 mg, 10 mg or even 50 mg or even 100 mg per day.

[0091] As demonstrated in the examples below and in a remarkable way, the extract according to the invention is compatible with the processes of preparing food products, in particular bread making, and gives the product obtained a GABA content compatible with health benefits, while not deteriorating the appearance and taste of said product, in particular bread. FIGURES

[0092] [Fig. 1]: Average specific volume (in cm³ / g) of the loaves obtained from different formulations:

[0093] Test 1; a "control" formulation corresponding to a dough without yeast extract

[0094] Test 2; the "control" formulation, in which a commercial yeast extract (Lesaffre - Springer Reveal 292 PW) has been added

[0095] Test 3; the "control" formulation, to which pure GABA has been added

[0096] Test 4; the "control" formulation, in which a yeast extract rich in GABA according to the invention has been added

[0097] [Fig.2]: Spiderweb pattern - main descriptors (Flour odor and aroma) White; Fermented aroma and scent; Cracker aroma and scent; Malty aroma and scent; Umami; Salty; Cracker crust aroma) and their scores (from 0 to 10)

[0098] + : significant difference (ANOVA, 5%) EXAMPLES OF ACHIEVEMENTS

[0099] The invention and its advantages will become clearer from the following embodiments, supported by the accompanying figures. These, however, are not intended to be limiting. 1 / Preparation of the active ingredient according to the invention

[0100] 1 / Preparation of the yeast extract A yeast extract of Saccharomyces cerevisiae was prepared and used at a concentration of between 30g / L and 300g / L.

[0101] 2 / Preparation of the bacterium carrying glutamate decarboxylase activity The bacterium Levilactobacillus brevis was cultured under suitable conditions on an MRS (Man Rogosa, Sharpe) type medium. The culture thus obtained is used as is or subjected to freeze-drying and then used in the step of converting glutamic acid to GABA.

[0102] 3 / Conversion of glutamic acid into GABA Bioconversion is carried out at a temperature between 30 and 40°C and a pH between 5 and 7. Inoculation of the bacterial extract with the bacterial wort initiates bioconversion. The bioconversion time is approximately 24 to 70 hours. The reaction is stopped when the conversion to GABA exceeds 90%, ideally complete. The resulting GABA-rich extract is then subjected to thermal inactivation and subsequently centrifuged.

[0103] The composition of the final extract obtained, in dry form, is given in the table below:

[0104] [Tables 1] Nitrogen (g%g dry) Total amino acids dry) Free amino acids (g%g dry) Acid | GABA (g%g dry; (g%g dry) 8.2 18 û 18 | 7.6

[0105] II / Use of the active ingredient according to the invention in a bread-making process 1 / Preparation of the different bread doughs:

[0106] The experiments were carried out on a "crusted bread" type formulation called "bastard", with a dough comprising water, T55 flour, salt, an improver containing ascorbic acid and baking enzymes, and "Blue Swallow" pressed yeast.

[0107] The flour and yeast used in these examples contain glutamic acid and GABA, but in small quantities, as shown in the table below:

[0108] [Table 2] Glutamic acid and GABA content in raw materials Ingredients: Glutamic acid, GABA (mg / 100g), Wheat flour "Biflor" 5.7, 43%, Yeast "Hirondelle bleue" 0.7, 5%, 2%

[0109] Different samples were prepared, according to the quantities described in Table 3 below: - a "control" formulation ("Control"; test 1) corresponding to a dough without yeast extract; - the "control" formulation, in which a commercial yeast extract (Lesaffre - Springer Reveal 292 PW) was added ("+ Springer Reveal 292 PW yeast extract"; test 2); - the "control" formulation, in which pure GABA (Sigma Aldrich -Gamma Aminobutyric acid, BioXtra > 99%) was added ("+ pure GABA"; test 3) - the "control" formulation, in which a GABA-rich yeast extract according to the invention has been added ("+ GABA-rich extract"; test 4).

[0110] [Table 3] Formulations tested 2 / Processing of different bread doughs:

[0111] The process implemented on the different pastes is the same and essentially comprises 3 steps: - mixing of ingredients; - dough rising (fermentation) for 110 minutes; - Cook for 30 minutes at 230°C. 3 / Analysis of bread products:

[0112] During the process, the level of CO2 production was measured by the Risograph system.

[0113] Bread volumes were measured with Volscan Profiler

[0114] For each sample, the GABA content of the final crumb was analyzed by HPLC. 3-1 / CO2 Production

[0115] The results obtained with the Risograph, after 110 minutes of fermentation, are shown in the table below:

[0116] [Table 4] Values ​​of CO2 produced in each sample Total CO2 (ni) Difference (%) Test 1 - Control 145.10 100 Test 2 - SpringerReveal Yeast Extract 292 PW 159.32 9.8 Test 3 - Pure GABA 147.92 1.9 Test 4 - GABA-rich extract 155.99 7.5

[0117] Pure GABA showed no impact on CO2 production, considering a fermentation time of 110 minutes. The addition of the yeast extract had a positive impact on CO2 production, with an increase of approximately 10% at 110 minutes. The GABA-rich yeast extract according to the invention also had a positive impact on CO2 production at 110 minutes (+7.5%). 3-2 / Bread volume

[0118] The results of the Volscan Profiler, allowing evaluation of the mean specific volume (in cm3 / g), are shown in [Fig. 1]. The results are expressed as a % difference between trials 2, 3 and 4 and the control (trial 1).

[0119] These results are consistent with those concerning CO2 production: loaves made with commercial yeast extract have the highest volumes, but the incorporation of a yeast extract according to the invention also has a positive effect. 3-3 / Sensory evaluation of the loaves obtained

[0120] Sensory evaluation was carried out on fresh bread by an expert panel:

[0121] - White light test

[0122] - Blind tasting, in a random order

[0123] - 6 expert panelists

[0124] - Quantitative (data analysis): rating of the intensity of each parameter on a 10-point scale (0 to 10). The parameters are: White flour odor and aroma; Fermented odor and aroma; Cracker odor and aroma; Malty odor and aroma; Umami; Salty; Cracker crust aroma.

[0125] The results of [Fig.2] reveal that: - The GABA-rich yeast extract according to the invention is significantly more malty; - The flavor profile of this bread is very different from the control. This solution provides significant flavors (malt and / or crackers) but no unpleasant taste. 3-4 / GABA content

[0126] The GABA content was measured in samples of crumb from the final loaf.

[0127] Free γ-aminobutyric acid (GABA) is quantified by reversed-phase HPLC-UV at 260 nm after derivatization using the AccQ-Tag Ultra kit (Waters, USA). An injection volume of 3 pL is injected into the chromatographic system equipped with a pre-column (XBridge BEH C18; 2.1 x 5.0 mm; 3.5 pm, Waters, USA) and a column (XBridge BEH C18; 3.0 x 150 mm; 3.5 pm, Waters, USA). The mobile phase consists of four lanes, which are respectively: Eluent A (A), Water / Eluent B 90 / 10 (V / V) (B), Ultrapure Water (C), and Eluent B (D).

[0128] The preparation of the different samples was carried out according to the following steps: - Dilution of 5 g of sample in 20 ml of Ultra pure water; - Homogenization of the solution by Ultra-turrax; - Addition of 5 ml of internal standard; - Centrifugation and filtration.

[0129] The results are presented in the table below:

[0130] [Table 5] GABA (free) content measured in the crumb and GABA (free) content calculated on the basis of the initial raw material input (theoretical) (mg / 10g) Measured GABA Difference (Theoretical x Measured) Test 1 - Control 2.6 1.2: 0 x - / 0 Test 2 - Springer Reveal Extract 292 PW 3.2 18 52% lest 3 - Pure GABA 24.2 19.3 -20% Test 4 - GABA-rich Extract .* - / 24.4 -12%

[0131] As expected, the standard (control) condition contains a low value of GABA in the final bread. The same is true for the bread with only Springer Reveal 292 PW yeast extract.

[0132] The theoretical calculated value (considering the GABA contribution of the different ingredients to the kneading) and the actual GABA content, measured within the framework of this invention, are similar.

[0133] The GABA content remains stable during breadmaking.

[0134] In the present invention, the incorporation of pure GABA (Sigma Aldrich - Gamma Aminobutyric acid, BioXtra > 99%) or of yeast extract rich in GABA makes it possible to obtain a bread product comprising more than 0.015 (corresponding to 150 ppm) or even more than 0.02% (corresponding to 200 ppm) by weight of GABA, the percentage by weight being expressed in relation to the weight of the product.

[0135] In conclusion, an extract according to the invention offers a natural ingredient which combines the benefits of GABA (for health) and a yeast extract (for breadmaking), and is also fully compatible with breadmaking processes.

Claims

Demands

1. Natural extract, preferably extracted from yeast, rich in gamma-aminobutyric acid (GABA), advantageously comprising at least 5% by weight of GABA, the percentage by weight being expressed in relation to the dry weight of the extract.

2. Extract according to claim 1, characterized in that it is low in glutamic acid, advantageously comprising less than 10% by weight of glutamic acid, advantageously less than 5%, even more advantageously less than 1%.

3. Extract according to claim 1 or 2, characterized in that it is a water-soluble extract, advantageously in dry form.

4. Extract according to any one of claims 1 to 3, characterized in that it is obtained from a yeast strain capable of synthesizing glutamic acid, for example from the species Saccharomyces cerevisiae.

5. Extract according to claim 4, characterized in that it is obtained by incubating the yeast extract in the presence of a glutamate decarboxylase (GAD) enzyme.

6. Extract according to claim 4, characterized in that it is obtained by incubating the yeast extract in the presence of a bacterium producing a glutamate decarboxylase (GAD) enzyme, advantageously carrying the GadA and / or GadB or even GadC genes, or of a lysate of such a bacterium.

7. Extract according to claim 6, characterized in that the bacterium belongs to the family of lactobacillaceae, advantageously chosen from the group consisting of: Levilactobacilus brevis, Lentilactobacilus buchneri and Lactococcus lactis.

8. Food product, in particular bread dough or bakery product, comprising an extract according to any one of claims 1 to 7, wherein the extract is advantageously present in an amount between 100 ppm and 3000 ppm, preferably between 150 ppm and 500 ppm, for example 200 ppm.

9. Food product according to claim 8 for use to improve the state of well-being and health, and / or to reduce the risk of disease in humans or animals.

10. Use of an extract according to any one of claims 1 to 7 in a breadmaking process, advantageously for the preparation of a GABA-enriched bread.

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