Germination of sprouts / seedlings and use of a mixture of dough with germinated sprouts / seedlings

Description

[0001] The present invention relates to the field of food production, namely a method for the sterilization of sprouts / germinates contaminated with microorganisms, in particular with pathogenic microorganisms (pathogens), suitable for the production of baked goods and / or their precursors, as well as their use.

[0002] Health-conscious consumers are increasingly aware of the importance of a nutrient-rich and balanced diet. Due to their high nutrient density and protein content, combined with low calorie density and high fiber content, sprouted seeds of edible plants (sprouts / seedlings) are becoming increasingly popular.

[0003] The benefits of consuming sprouted seeds as food and medicine were recognized in East Asia, particularly in China, as early as 5000 years ago. With the spread of Asian cuisine and medicine, sprouts (also known as shoots or sprouting grains) from cereals, legumes, and oilseeds became known in the Western Hemisphere from the 18th century onward. Only relatively recently have these sprouts been used in the commercial production of baked goods.

[0004] "Disinfection" within the meaning of the present invention is understood to mean the process of bringing microorganisms into a state in which they are no longer capable of division and growth and / or are infectious, through appropriate treatment.

[0005] The terms "seed, seed or grain" are used synonymously for the purposes of the present invention.

[0006] For the purposes of the present invention, "plant germs" or "germs" are understood to be all embryos (plant germs) resulting from the fertilization of plant egg cells, including shoot and leaf germs (plumula) and root germs (radicle).

[0007] "Seedlings" within the meaning of the present invention are understood to be all seeds from the first germination stage onwards, the so-called soaked seed (water-soaked seed), in particular all seeds whose plant germs are in the active growth phase and have already formed root and seedlings that are still dependent on the nutritive tissue of the seed (endosperm).

[0008] "Sprouts" within the meaning of the present invention are understood to be all shoot axes of seedlings growing out of the germinating seeds or already grown out, some of which are already covered with leaves.

[0009] In botany, germination refers to the first stage of ontogeny in seeds. The nutritionally beneficial nutrients and vital substances that arise during plant germination have brought these seedlings / sprouts into focus in recent years for the commercial production of baked goods, where they are increasingly used.

[0010] Before germination begins, the seed, as a storage organ, is in a state of dormancy characterized by very low metabolic activity. Germination is initiated by the supply of oxygen, water, and a suitable germination temperature. At the beginning of the germination process (swelling), the grain is given time to swell (soaking); the water content in the seed rises to approximately 70%, and the dormant embryo begins to develop. During this phase, metabolism increases, and numerous biochemical transformations take place. For example, the moist seed coat becomes breathable, it splits open, and rhizomes emerge from the seedling, followed by the shoot from the cotyledon sheath, which will form the developing plant.By activating existing enzymes and / or synthesizing new ones, plant substances in the seed are transformed and the seed contents are partially broken down, so that they can be used by the growing seedling to generate energy.

[0011] During germination, various processes of breakdown, transformation, and synthesis take place. The increased metabolism during germination is also associated with an increased synthesis of nutrients, such as vitamins, especially vitamin C. Furthermore, there is a noticeable increase in the quantity of essential amino acids, particularly lysine. The complex starch is broken down into less complex polysaccharides and monosaccharides in a process known as predigestion. Compared to the original seed, this predigestion results in an increase in essential fatty acids and an overall improvement in digestibility.

[0012] Consumers, who are increasingly aware of the higher and more easily digestible nutrient content of sprouts / germinated seeds, are also becoming more aware of this, which makes the use of sprouts / germinated seeds increasingly interesting for the food industry.

[0013] Once the sprouts have reached approximately the length of grains, they can be harvested or used for further purposes. However, excessively long germination times can lead to vitamin loss and a negative impact on flavor, which should be avoided.

[0014] The numerous biochemical processes during germination increase the digestibility of the sprouts / germinations and lead to the formation of various physiologically valuable secondary plant compounds. For example, the germination process results in a significant increase in vitamins B1, B2, B6, C, E, biotin, and folic acid. While the baking process inevitably leads to a loss of overall nutrient content, adding sprouts / germinations to the final product can still generate a significantly higher vitamin content, which can be verified using appropriate measurement methods, compared to baked goods made with unsprouted grains of the same variety.

[0015] As a natural product, a certain spectrum of microorganisms and pathogens is normal and naturally occurring. However, when cultivating sprouts / seedlings from seed, particular care and attention are required regarding pre-contamination with pathogens, such as microorganisms, especially pathogenic microorganisms. Natural pre-contamination with pathogens, especially microorganisms such as bacteria, which are naturally ubiquitous, can occur particularly through the air, rain, or fertilization with manure, through the harvesting itself (e.g., through contaminated harvesting equipment or transport containers), through drying in storage areas, through the conditions during storage, transport, and human and machine processing, e.g., through smear infection.

[0016] Typically, oxidative substances such as hydrogen peroxide (H₂O₂) or hypochlorite salts, such as sodium hypochlorite (NaClO), are used to disinfect the seed surface. These are usually applied as aqueous solutions before the seeds have swelled. The seeds are exposed to the oxidative substances in sterile containers for the appropriate exposure time. This is followed by several rinses with sterile water, possibly with additional or intermediate rinses, e.g., with alcohol. In some cases, this process is carried out using ultrasound for mechanical cleaning of the seeds.

[0017] H2O2-based disinfectants often have silver ions added to enhance their microorganism-killing effect, which permanently pollutes wastewater and thus the environment.

[0018] Disadvantages of chlorine-based disinfection include the fact that some of the chlorine forms organic halogen compounds, which are difficult to break down and consequently increasingly pollute wastewater and thus the environment.

[0019] In addition, a purely physical method has recently become available for disinfecting seeds, utilizing the germicidal effect of accelerated electrons. This method was developed by the Fraunhofer Institute for Organic Electronics, Electron Beam and Plasma Technology FEP. However, in Germany, according to the Food Irradiation Ordinance of December 14, 2000, and EU Directives 1999 / 2 / EC and 1999 / 3 / EC, the treatment of foodstuffs other than dried aromatic herbs and spices with accelerated electrons is not permitted.

[0020] A particular danger therefore exists during seed germination, which usually occurs without the presence of disinfectants. Due to the warm, humid conditions prevalent during germination, which are essential for the germination of cereal seeds, the risk of proliferation increases, even from a small initial population of pathogenic microorganisms, especially bacteria, salmonella, molds, viruses, and yeasts, which thrive, multiply, and spread very well under such germination conditions.

[0021] In seedling production, it is common practice to wash harvested sprouts / seedlings to remove any remaining adhering substrate and reduce the surface pathogen load. For example, citric acid is added to the wash water for mung bean or soybean sprouts / seedlings to give them a more appealing, lighter appearance for the customer. The citric acid also lowers the pH of the wash solution, reducing the number of germs on the sprouts / seedlings in contact with it. The sprouts / seedlings are then rinsed from the wash solution and stored in a cold storage facility at a maximum of +6°C until further processing. However, this process does not guarantee the complete and effective removal of pathogens or the sterilization of the product, especially of resistant pathogens such as EHEC or Salmonella.

[0022] Document WO 96 / 23528 describes a method for the surface disinfection of plant material or plant tissue material not present as a permanent form (seed) and intended for vegetative plant propagation by means of combined treatment of the plant tissues with ultrasound in the presence of an aqueous solution of oxidative disinfectant, such as sodium hypochlorite, in which the plant tissue in question is located.

[0023] However, even with sufficient hygienic working methods up to the germination of the grain seeds, the microbiological hygiene and shelf life of foodstuffs, such as baked goods, pose considerable problems for the manufacturers of these goods during further processing, especially since the consumer wants to avoid the use of chemical disinfectants on the seeds.

[0024] Contamination of the sprouts / germinated seeds used in baked goods with microorganisms can occur, particularly during packaging and transport. Besides the air, equipment, machinery, and personnel are the most significant potential vectors for pathogens to reach the sprouts / germinated seeds.

[0025] In fact, since a major epidemic with a so-called enterohemorrhagic Escherichia coli (EHEC) strain that occurred in Japan in 1996, sprouts have been considered possible vectors of pathogenic germs, such as EHEC.

[0026] However, since the major hemolytic uremic syndrome (HUS) epidemic, which occurred primarily in Germany in May 2011 but also in other parts of Europe, and during which sprouts were considered suspected carriers of the pathogen, which is life-threatening to humans, according to the report at the time by the Federal Institute for Risk Assessment (BfR), the Federal Office of Consumer Protection and Food Safety (BVL), and the Robert Koch Institute (RKI), there has been considerable uncertainty among the population regarding the safe consumption of sprouts / germinated seeds, especially in their raw form. At that time, a total of 855 people contracted HUS and 2,987 contracted gastroenteritis, with 53 people dying from the infection.

[0027] The cause was assumed to be so-called enterohemorrhagic Escherichia coli (EHEC), a pathogenic strain of the naturally occurring intestinal bacterium. Escherichia coli ( E. coli ).

[0028] E. coliIt belongs systematically to the family of Enterobacteriaceae, which also includes the "classic" diarrheal pathogens, such as Salmonella and Shigella (dysentery pathogens). In contrast to the naturally occurring non-pathogenic bacteria... E. coli EHEC strains are pathogenic compared to strains that perform important nutritional functions in the intestine, such as nutrient processing and defense against pathogens as part of the normal intestinal biota. E. coli Strains characterized by the production of toxins. These so-called Shiga or Verotoxins are potent cytotoxins that can cause serious intestinal diseases in humans, including HUS.

[0029] EHEC bacteria are naturally found in the intestines of ruminants, such as cattle, goats, or sheep, and are excreted in the animals' feces. They can be transmitted to humans directly or indirectly via fecal-oral contact and can cause life-threatening illnesses, such as the described HUS or other diseases of the gastrointestinal tract.

[0030] Furthermore, there is a potential risk of salmonella infection from sprouts / germinates contaminated with salmonella. Salmonella poisoning, also known as salmonellosis, is typically endemic and, along with acute respiratory illnesses, is one of the most common infectious diseases in humans. In recent years, for example, 90% of foodborne infections in Germany have been attributed to salmonella. Inadequate refrigeration of food, in particular, promotes the spread of salmonella. This can lead to rapid bacterial growth in food, especially during the summer, until a critical infectious dose is reached. The infectious dose is a measure of the amount of pathogen required to cause illness. Furthermore, the individual susceptibility of consumers is a crucial factor in determining the level of the infectious dose.The resistance of salmonella to harmful environmental influences and its high reproduction rate have contributed to salmonella diseases being the most common foodborne illness in humans.

[0031] It is noteworthy that, unlike many other bacterial infectious diseases, only a few pathogens (depending on the source, just under to just over 100 EHEC bacteria; Salmonella: 10,000, in fatty foods about 100) are sufficient to cause illness, especially in particularly susceptible individuals such as infants, young children and the elderly; that is, the infectious dose for EHEC, but also for Salmonella, is comparatively low.

[0032] Heating the food to a core temperature of approximately 70°C for about 2-10 minutes kills pathogenic organisms such as EHEC and Salmonella and inactivates any toxins that may be present. In contrast, EHEC and Salmonella are relatively resistant to other environmental influences, such as cold (even deep freezing), temporary acidification (like the natural acidification caused by stomach acid during the passage of orally ingested food through the stomach), or dehydration. Therefore, baked goods containing sprouts / germinated seeds that may be contaminated with pathogenic organisms are safe from an infectious standpoint, as a sufficient core temperature is reached within the necessary timeframe during baking. Likewise, any bacterially produced toxins are heat-inactivated by baking at a sufficiently high and long core temperature (as described above).Therefore, microbial spoilage is the decisive factor here, leading to a significant reduction in nutrients and vital substances in the sprouts / germinations affected by the microorganisms, and is also undesirable from a hygienic point of view for the consumer.

[0033] The situation is different for the use of unbaked and insufficiently cooked baked goods or pre-ferments and sourdoughs, e.g. as an addition to muesli or cornflakes or baked goods or toppings that are not cooked all the way through to the core.

[0034] Equally problematic and highly problematic can be hygiene issues, especially due to the spread of smear infections, when sprouts / germinates contaminated with microorganisms are used in the production facilities of so-called baked goods with unbaked fillings, such as custard pastries, or in facilities where baked goods and cream products, e.g., profiteroles or cream cakes, are manufactured.

[0035] Nevertheless, as already mentioned, sprouts / germinated seeds are among the highly perishable foods whose production process carries the possibility of rapid microbial spoilage and the risk of contamination with pathogenic germs, such as E. coli, Caused by Listeria, Salmonella or viruses such as norovirus or hepatitis A virus.

[0036] Even with typically cool storage, pre-packaged sprouts contaminated with microorganisms can multiply rapidly within a few days, resulting in a very high microbial load. If contamination with pathogenic organisms such as EHEC or Salmonella is present, there is an increased risk of serious, potentially fatal, illness in humans.

[0037] Against this background, the invention is based on the objective of at least partially overcoming the disadvantages known in the prior art of the transfer of contaminants and / or pathogenic microorganisms to plant products to be processed. In particular, it is to be recognized as a success of the inventors of the present invention that they provide a method for the sterilization of sprouts / seedlings contaminated with microorganisms without the use of chemical and / or oxidative seed / seedling disinfectants known in the prior art, so that such sprouts / seedlings have a hygienically sound and defined quality for the production and further processing in food and / or baked goods. Document DE 69517442T2 discloses a method for the fermentation of cereal germs with lactic acid bacteria, which are subsequently used for baked goods.

[0038] The inventive method for sterilizing sprouts / germinates contaminated with microorganisms is particularly suitable for the production of baked goods and / or their pre-products, in particular pre-doughs and / or sourdoughs, using water and at least one grain milling product, which is acidified by the addition of inoculant containing lactic acid bacteria and optionally yeast.

[0039] The method according to the invention is characterized by the features of claim 1.

[0040] The inventors have surprisingly discovered that the inventive method efficiently sterilizes sprouts / germinates, for example, in pre-products of baked goods, particularly in preferments and / or sourdoughs. Only a coarse washing process is required to remove substrate residues from the sprouts / germinates, rather than an extensive washing process with multiple rounds / steps. This significantly simplifies production and improves quality in baked goods because the sterilization process takes place directly in the pre-products, especially in preferments and / or sourdoughs. Recontamination is thus essentially eliminated, and process and cleaning water with the corresponding additives are saved, thereby conserving energy and protecting wastewater and the environment.

[0041] According to the invention, the inoculum used in the process consists of lactic acid bacteria and / or other microorganisms, or can be a sourdough starter and / or finished sourdough. The sourdough thus serves as a sterilization medium in which pathogenic microorganisms are displaced, overgrown, or inactivated by acidification. Hereinafter, the general term "dough" is used synonymously with sourdough, sourdough starter, and sourdough-containing mixtures of cereal products and water.

[0042] After the sterilization process according to the invention, the resulting acidified mixture can be baked directly as dough with sterilized sprouts / germinates. This simplifies the manufacturing process for baked goods containing sterilized sprouts / germinates.

[0043] In particular, the germ-free mixture according to the invention is characterized by the fact that this mixture, in other words, the dough with sprouts / germinations is in pasty form and has been advantageously improved by the use of the hydrocolloids described below.

[0044] The use of at least one gel-forming hydrocolloid in a specific ratio to the remaining components significantly determines the flow properties of the mixture and simultaneously ensures an even distribution of the sprouts / germinations in the dough.

[0045] It has been shown that, particularly in the pasty mixture, the even distribution of the sprouts / seedlings and simultaneously the even distribution of the added lactic acid bacteria and / or other microorganisms is enabled, resulting in significantly improved sterilization, since there are no "dead spaces" in which the lactic acid bacteria are not or insufficiently active, e.g. due to the clumping of sprouts / seedlings.

[0046] A further advantage is that this dough with sprouts / germinates remains pumpable, ensuring a stable and even distribution of the sprouts / germinates within the dough even after storage and transport. According to the present invention, the term "pumpable" implies that the dough with sprouts / germinates is still fluid, and thus potentially so viscous that there would be a risk of settling and consequently a loss of even distribution of the sprouts / germinates. From the customer's perspective, this is particularly crucial for maintaining consistent dough consistency and quality. It also ensures that downstream processing plants can pump out the dough using a low, constant pumping rate. This is especially important with dough containing sprouts / germinates that are not permanently evenly distributed, which can occur during storage or transport.The transport process, caused by gravity, would cause the dough to sink to the bottom of the dough container and accumulate there. This may not be the case, as a higher pumping capacity would be needed to pump out the more consistent dough fraction at the bottom of the dough container, which is due to the higher number / density of accumulated sprouts / germination in the dough at the bottom of the dough container.

[0047] According to one embodiment, the sprouts / seedlings according to the invention are obtainable from malted seed and / or leaf sprouts / root sprouts from seed, wherein the malted seed and / or leaf sprouts / root sprouts are separated and dried after germination.

[0048] "Malted seed" within the meaning of the present invention is seed that has already germinated but still has certain seed reserve substances.

[0049] By separating the malted seed and the leaf / root sprouts from each other and subsequently drying them separately, the sprouts / seedlings according to the invention achieve a particularly high quality.

[0050] The varieties of sprouts / germinated seeds to be used in the inventive process are wheat, spelt, rye, barley, oats, rice, millet, quinoa, lettuce, onion (seed), chickpeas, lentils, leeks, anise, caraway, fennel, fenugreek, corn, beans, peas, soybeans, alfalfa, radish, daikon radish, cress, nasturtium, chia, broccoli, arugula, alfa-alpha, amaranth, pumpkin, mustard, sesame, flaxseed, sunflower, and tomatoes, or mixtures thereof. The added, at least one gel-forming hydrocolloid, is preferably a hydrocolloid selected from the group consisting of xanthan gum, alginate, carrageenan, hydroxypropyl methylcellulose, inulin, and highly esterified pectin, or mixtures thereof. The gel-forming hydrocolloids according to the invention are characterized by the fact that they form gels when cool without a heat-mediated activation step.

[0051] According to a preferred embodiment, the non-heat-resistant hydrocolloid is a pectin and / or inulin or mixtures thereof.

[0052] In addition to the gel-forming hydrocolloids according to the invention, it may be appropriate to use thickeners such as starch, guar gum or locust bean gum.

[0053] Due to the advantageous and modulatable flow properties of the mixture adjusted using gel-forming hydrocolloids, it has also become possible, according to a further embodiment, to apply this mixture to the sprouts / seedlings by spraying. It has been found that this allows for a finer distribution of the mixture of lactic acid bacteria and / or other microorganisms on the sprouts / seedlings. At the same time, it was shown that this fine distribution forms a fully enclosing layer, which also exhibits a correspondingly advantageous and improved germination effect.

[0054] The advantageous and modulatable flow properties of the pasty pumpable dough (germination mixture) according to the invention are adjusted via a specific ratio of the at least one gel-forming hydrocolloid to the total mass of the pasty dough (essentially the mixture of water, grain milling product and inoculation material).

[0055] According to a further embodiment, wherein the at least one gel-forming hydrocolloid is selected from the group consisting of xanthan gum, alginate, carrageenan, hydroxypropyl methylcellulose and esterified pectin, i.e., with a degree of esterification > 50% or mixtures of the aforementioned, the gel-forming hydrocolloid is used in the specified ratio of approximately 0.2 to approximately 5%, preferably approximately 0.5 to approximately 3%, and particularly preferably approximately 0.6 to approximately 2% based on the total mass of the pasty dough.

[0056] According to a preferred embodiment, the gel-forming hydrocolloid according to the invention is inulin, and the specific ratio of inulin to the total mass of the pasty dough is approximately 2–12%, preferably approximately 4–10%, and particularly preferably approximately 5.3–8.7%.

[0057] The incubation of the mixture of dough and sprouts / germinated seeds takes a total of approximately 1 to 10 days, preferably approximately 2 to 8 days, and particularly preferably approximately 3 to 5 days or approximately 2 to 3 days. Sterilization takes place at an acidification pH of ≤ approximately 4.3, preferably approximately pH 3.0 to 4.5, and particularly preferably approximately pH < 4.2, and at a temperature of approximately 12 to 30°C, preferably approximately 15 to 25°C, and particularly preferably approximately 18 to 22°C.

[0058] According to the invention, the sterilization of the mixture of sprouts / germinates contaminated with microorganisms is achieved without the addition of additional acidifying agents such as lactic acid, acetic acid, citric acid or other organic or mineral acids.

[0059] The inoculation material according to the invention contains an adapted mixed flora of at least one strain of homo- and / or heterofermentative lactic acid bacteria, wherein the strain of homo- and heterofermentative lactic acid bacteria is selected from the strains L. acidifarinae, L. acidophilus, L. alimentarius, L. amylovorus, L. brevis, L. buchneri, L. cellobiosus, L. coleohominis, L. collinoides, L. crispatus, L. crustorum, L. curvatus, L. delbrueckki, L. diolivorans, L. farciminis, L. fermentum, L. fructivorans, L. frumenti, L. gallinarum, L. gasseri, L. hammesii, L. helveticus, L. hilgardii, L. homohiocchi, L. johnsonii, L. kefiri, L. kimchi, L. kunkeei, L. linderi, L. mali, L. mindensis, L. mucosae, L. nagelii, L. nantensis, L. namurensis, L. nodensis, L. oris, L. panis, L. paraalimentarius, L. parabuchneri, L. paracasei, L. pentosus, L. perolens, L. plantarum, L. pontis, L. reuteri, L. rossiae, L. sakei, L. sanfranciscensis, L. secaliphilus, L. siliginis, L. spicheri, L. vaginalis, L. zymae, ua Lactococcus lactis, Leuconostoc citreum, Lc. argentinum, Lc. suntoryeus, L. gelidum, Lc. mesenteroides, Pediococcus acidilactici, P. damnosus, P. parvulus, P.pentosaceus, Weissella cibaria, Weissella confusa, Weissella kandleri, Weissella paramesenteroides, Weissella viridescens. and mixtures thereof.

[0060] According to one example, the mixture is enriched with a sourdough starter culture, which preferably - but not exclusively - consists of lactobacilli selected from the group Lactobacillus plantarum, L.fermentum, L.paracasei, L.paralimentarius, L.helveticus, Leuconostoc argentinum and Saccharomyces pastorianus contains.

[0061] The lactic acid bacteria must be suitable for generating the necessary low pH for sterilization. Suitable lactic acid bacteria are known to those skilled in the art and are included within the scope of the invention.

[0062] Optionally, the inoculation material according to the invention can additionally contain at least one acid-tolerant yeast strain.

[0063] The at least one yeast strain must be suitable for tolerating the necessary low pH for sterilization. Suitable yeast strains are known to those skilled in the art and are included within the scope of the invention. In particular, one yeast strain is selected from the strains Candida milleri (Kazachstania milleri), Candida humilis, Kazachstania exigua, Saccharomyces cerevisiae, Debaryomyces hansenii, Dekkera bruxellensis, Kazachstania unispora, Kluyveromyces lactis, S. bayanus, Saccharomyces pastorianus, Torulaspora delbrueckii, T. pretoriensis, Wickerhamomyces anomalus, Pichia anomala, Hansenula anomala, Pichia kudriavzevii, Issatschenkia orientalis, Candida krusei and mixtures thereof.

[0064] According to one embodiment of the method according to the invention, the method kills approximately 98% of the pathogenic microorganisms that contaminate the sprouts / seedlings. A reduction, inactivation, and / or killing of 99%, and furthermore of 95%, 90%, 85%, 80%, and 75% of the microorganisms on the contaminated sprouts / seedlings, particularly the microorganisms of the following, is entirely sufficient and achievable in any case by using the present invention. E. coli Strains of EHEC and / or Salmonella. In particular, the method according to the invention aims to and achieves a reduction of the respective pathogenic microorganisms to values ​​below the respective critical infectious dose for humans.

[0065] Even after just one day of exposure in the sourdough starter according to the invention, the number of microorganisms that contaminate the sprouts / seedlings, in particular the microorganisms of the E. coli Strains of EHEC and / or Salmonella were reduced by approximately 3 log steps, in particular to approximately 0.2% of the initial number at day 0, i.e., more than approximately 99.8% of the initial number of microorganisms was eliminated.

[0066] The inventive method, in particular, does not require the application of heat to inactivate the unwanted microorganisms, and especially not a pasteurization step. This ensures that the lactic acid bacteria in the inoculation material remain alive and continuously active. Only during the extended sterilization process, lasting several days, can the mixed flora be inactivated or killed by the low pH itself.

[0067] The sprouts / germinated seeds obtained using the inventive method are also described. They possess the required hygienically impeccable quality necessary for use in, for example, baked goods.

[0068] According to the description, it is also possible to sterilize undried sprouts / germinations, i.e., fresh or raw sprouts / germinations, using the inventive method and then process them further, for example, for baking or for raw consumption. This is particularly advantageous because fresh raw sprouts / germinations cannot currently be sterilized to a sufficiently hygienic standard using the sterilization methods available in the prior art.

[0069] The use of the inventive method, e.g. by spraying, is particularly preferred for the sterilization of sprouts / germination seeds for raw consumption. The sterilized sprouts / germination seeds can then be washed for raw consumption.

[0070] Furthermore, the present invention relates to the use of an acidified mixture of dough with sterilized sprouts / germinates produced according to the inventive method for the production of baked goods.

[0071] Baked goods made from a sourdough mixture of dough with sterilized sprouts / germinates are also part of the description. List of characters:

[0072] Figure 1 :Significant reduction in the viable count of EHEC and Salmonella in the sourdough mixture according to the invention, consisting of sourdough (40%), inulin (8%), and sprouts / germinated seeds (52%), during incubation at different times after preparation of the sourdough mixture (growth curve); (black solid line) EHEC culture according to the invention; (dark gray solid line) Salmonella typhimurium of the invention - Approach; (light gray dashed line) Control approach (E. coli, Cultivated under standard ideal conditions (calculated literature values). Measured values ​​according to Table 1; x-axis: days after preparation of the sourdough starter, y-axis: live cell count [colony-forming unit (CFU) / g], log plot Figure 2 and 3 :Photographic comparison of the change in the uniform distribution of sprouts / germinates mixed in the sourdough mixture and also the retention of water during the resting / storage of the recipe according to the invention (indicated by "Experiment 2"; right half of the photograph) in comparison to a prior art recipe (control, indicated by "Experiment 1"; left half of the photograph) Examples Example 1: Production of the inventive mixture from sourdough with inulin and cereal sprouts / germinates contaminated with EHEC or Salmonella

[0073] For the sourdough starter, milled grain products and water were mixed in a 1:1 ratio with the inoculant to form a dough-like mass (sourdough). The inoculant dosage was 10% of the amount of milled grain products. Fermentation then took place at 26-28 °C for approximately 24 hours.

[0074] Next, an inulin compound containing 8% inulin was added to the sourdough. Immediately afterwards, the wheat sprouts / germinated grains were added and thoroughly mixed into the sourdough.

[0075] The prepared mixture of sourdough, hydrocolloid (inulin) and cereal sprouts / germinates was further incubated with intermittent stirring and adjusted to a pumpable final consistency.

[0076] The example recipe is: Sourdough: 40% Inulin: 8% Sprouts / germinated seeds: 52% Example 2 Significant reduction in the number of live EHEC and Salmonella bacteria

[0077] The procedure for determining the influence of the sourdough starter according to the invention (so) on the viable cell count of cereal sprouts / germinates pre-contaminated with EHEC or Salmonella was as follows: First, pre-cleaned wheat grains were soaked in drinking water at 18 °C for 12 hours. Excess water was then removed, and germination was carried out at approximately 20 °C. Water was added as needed. After 3 days, the sprouts / germinates were placed in a suspension of E. coli O157:H7 was converted to achieve a cell count of 105 colony-forming units (CFU) / g. A corresponding approach was used instead of with Escherichia coli O157:H7 with a Salmonella strain (Salmonella typhimurium)The following steps were carried out: Sprouts / germinates and sourdough (dough yield 200) were mixed in a 1:1 ratio, divided into individual portions (50 g units), and packaged in polyethylene bags. At the beginning of the experiment (0 days) and at various subsequent time points, live cell counts were determined using the spatula method. E. coli O157:H7 were grown on Sorbitol McConkey (SMAC) agar and the number of CFU / g was recorded. Salmonella typhimurium accordingly on XLD agar.

[0078] For very low bacterial counts, an enrichment of EHEC / Salmonella was carried out in order to also detect damaged cells.

[0079] At the beginning (day 0) of the experiment, the measured initial output number was E. coliO157:H7 2.5 * 10⁵ < CFU / g. This number was significantly reduced to 5.0 * 10² < CFU (day 1) and to 1.0 * 10² < CFU / g (day 2) after incubation for one day in the sourdough according to the above recipe (see Figure 1 , Table 1). After 5 days, the EHEC bacteria in the sample were almost completely reduced to 5.0 * 10 1< CFU / g (day 5). After 7 days of incubation, only slightly more than 1.0 * 10 1< CFU / g were detectable (day 7). After 13 days of incubation (day 13), the last measurement was taken. No further EHEC enrichment could be detected. Derived from the growth curve ( Figure 1 However, it can be seen that even with an incubation period of less than 7 days, all living EHEC pathogens were reduced to undetectable quantities, i.e., completely eliminated, and could no longer be detected even after enrichment.

[0080] The results show that after just one day of exposure in the sourdough starter according to the invention (day 1), the EHEC viable count was reduced by approximately 3 log units to approximately 0.2% of the initial count at day 0, i.e., more than approximately 99.8% of the initial number of EHEC microorganisms was eliminated. As described above, the infectious dose for EHEC, depending on the source, is just under 100 to just over 100 EHEC pathogens to cause a disease-causing infection. After two days of incubation according to the method according to the invention, the EHEC pathogens were reduced to a total of approximately 100 (1.0 * 10²) bacteria, i.e., very advantageously, possibly below or close to the critical infectious dose threshold.

[0081] This is all the more surprising since it is known from the literature that the number of E. coli under standard ideal conditions, it grows exponentially: According to literature, the generation time of E. coli,EHEC also divides under optimal incubation conditions for approximately 20 minutes, meaning that an EHEC bacterium divides every 20 minutes under optimal conditions. With an initial count of 2.5 * 10⁵ < CFU / g, this would theoretically result in approximately 1.18059 E + 27 CFU / g after one day, 5.57519 E + 48 CFU / g after two days, and so on (see Figure 1 However, this is only a theoretical value, since no more than a maximum of 1013 cells can fit in 1 g. The inventive method, using the inventive composition, not only stops this fatal exponential "growth boom" of pathogenic microorganisms, but also significantly and advantageously reduces their number in a relatively short time to very low values ​​relative to their initial number, down to a number below the critical infectious dose and even lower.

[0082] The parallel approach in the sourdough starter according to the invention instead E. coli O157:H7 Salmonella typhimuriumThe initial number of [unclear text] used at the beginning of the experiment (day 0) was [unclear text]. Salmonella typhimurium of 5.0 * 10⁴ < CFU. The infectious dose described in the literature for Salmonella is approximately 10,000 bacteria; the initial bacterial count was therefore approximately five times the infectious dose. After a corresponding incubation of the culture for 1 day, the number of CFU / g Ω was already reduced by more than 3 log units to < 1.0 * 10¹ < CFU / g (day 1), thus significantly below the infectious dose. After 2 days, it was possible to S. typhimuriumSalmonella bacteria could only be detected after enrichment (day 2, detectable but not countable); after 5 days (day 5), the enrichment was negative (see Table 1). This means that it can be assumed that by day 5 at the latest, all initially present Salmonella bacteria were killed by incubation in the sourdough starter according to the invention. This represents a very promising positive finding with regard to hygienically safe sprouts / germinates, especially for the food industry.

[0083] Without being bound to the theory, it is assumed that the difference between the result with the Salmonella strain used and the EHEC strain can be attributed to a generally known higher acid tolerance of EHEC pathogens compared to Salmonella despite the same treatment.

[0084] Nevertheless, the inventors of the present invention have surprisingly and remarkably discovered that the generally quite acid-resistant Salmonella strain Salmonella typhimurium and the highly acid-resistant EHEC strain E. coliO157:H7 can be significantly reduced to viable cell counts below the respective pathogen-specific critical infectious dose or to undetectable viable cell counts in a relatively short period of time using the inventive method and the inventive composition. Accordingly, it can be assumed that other microorganisms, in particular acid-resistant pathogenic microorganisms such as other Salmonella strains and EHEC strains, but also Listeria or viruses such as noroviruses or hepatitis A viruses, can also be advantageously eliminated or reduced to low numbers below the respective pathogen-dependent infectious dose using the inventive method and the inventive composition. Table 1: Salmonella typhimurium E. coli Determination of the viable cell count of the dough mixtures according to the invention with contaminated or O157:H7 (EHEC) contaminated sprouts / germinates at various incubation times. Day 0 denotes the day immediately following the preparation of the respective test mixtures. day Salmonella typhimurium: Viable cell count on XLD agar E. coli O157:H7 (EHEC): Viable cell count on SMAC agar 0 5.0 * 10⁴ < CFU / g 2.5 * 10⁵ < CFU / g 1 Approx. 1.0 x 10¹ < CFU / g Approx. 5.0 x 10² < CFU / g 2 Enrichment positive Approx. 1.0 x 10² < CFU / g 5 Enrichment negative Approx. 5.0 x 10¹ < CFU / g 7 - Approx. 1.0 x 10¹ < CFU / g 13 - Enrichment negative Example 3 The composition (recipe) according to the invention containing inulin enables a better even distribution of the sprouts / germinations mixed therein and also a better retention of water than prior art recipes.

[0085] In a further experiment, the inventive recipe described above, i.e., sourdough 40%, inulin 8%, sprouts / germinated seeds 52%, was prepared, mixed in equal proportions, and then left to stand. The even distribution of the sprouts in the composition and the water retention in the sourdough mixture were visually observed and evaluated over time. A corresponding "prior art" composition without inulin served as a control.

[0086] Over time, the control approach (in Figure 2 (Indicated with "Attempt 1"; left half of the photograph) a watery solution was applied as a top layer over the dough mixture. After resting for 4 days, this layer had already grown to approximately 0.6 cm in the control batch (see Figure 2 In sharp contrast to this, the approach of the composition according to the invention (in Figure 2(indexed with "Attempt 2"; right half of photograph) homogeneous, i.e. the aqueous solution remains retained in the dough itself after the dough mixture has rested for 4 days (see Figure 2 ) and 5 days (see Figure 3 ).

[0087] In addition to the settling of a supernatant, the reduced uniformity of the sprouts / germinations in the dough can be observed in the control batch; that is, the sinking of the sprouts / germinations due to gravity is evident in the form of an increasing accumulation of sprouts / germinations towards the bottom of the container (at least to some extent), which was absent in the batch according to the invention. Therefore, in the composition according to the invention, the sprouts / germinations remained uniformly distributed in the dough, in stark contrast to the control batch. Since, as shown in Example 2, Figure 2Since the proportion of sprouts / germinations relative to the total composition of each batch was very high (52%), the aforementioned improved effect of the even distribution of sprouts / germinations in the dough according to the present invention was not very pronounced compared to the control batch. Therefore, further experiments with various lower proportions of sprouts / germinations relative to the total composition of the batch were subsequently investigated for this effect. These experiments showed the difference in the distribution of sprouts / germinations between the control batch and the corresponding batch according to the invention even more clearly (not shown).

Claims

1. A method for sterilizing sprouts / seedlings contaminated with microorganisms, especially pathogenic microorganisms, for the production of baked goods and / or their precursors, especially pre-doughs and / or sourdoughs, using water and at least one grain mill product, which is acidified by adding inoculum containing lactic acid bacteria, the inoculum optionally also containing yeast, characterized in that sterilization is achieved by the following steps: - mixing a paste-like dough from water, grain mill product, inoculum and at least one gel-forming hydrocolloid in a specific ratio; - mixing the resulting pumpable paste-like dough with the sprouts / seedlings, thereby producing a mixture of dough and sprouts / seedlings distributed essentially uniformly in a colloidal form; - incubating the mixture of dough and sprouts / seedlings to sterilize the sprouts / seedlings, the incubation being carried out for a total of 1-10 days and at a pH of 3.0-4.5 and a temperature of 12-30°C; - providing the mixture of dough with sterilized sprouts / seedlings.

2. The method of claim 1, characterized in that the sprouts / seedlings are from malted seeds (seed bodies) and / or plumules / radicles from seeds, wherein the malted seeds and / or plumules / radicles are separated and dried after germination.

3. The method of one of claims 1 or 2, characterized in that the varieties of sprouts / seedlings are wheat, spelt, rye, barley, oats, rice, millet, quinoa, lettuce, onion (seeds), chickpeas, lentils, leeks, aniseed, caraway, fennel, fenugreek, corn, beans, peas, soybeans, alfalfa, radishes, cress, nasturtium, chia, broccoli, arugula, alfalfa, amaranth, pumpkin, mustard, sesame, linseed, sunflower, and tomatoes, or mixtures thereof.

4. The method of any preceding claim, characterized in that prior to the mixing step, there is a step of applying the paste-like, pumpable dough by spraying onto the sprouts / seedlings.

5. The method of any preceding claim, characterized in that the at least one gel-forming hydrocolloid is selected from the group consisting of xanthan, alginate, carrageenan, hydroxypropyl methylcellulose, inulin, and highly esterified pectin, or mixtures thereof.

6. The method of claim 5, characterized in that the specific at least one gel-forming hydrocolloid is selected from the group consisting of xanthan, alginate, carrageenan, hydroxypropyl methylcellulose, and highly esterified pectin or mixtures thereof, and wherein the gel-forming hydrocolloid is used in the specific ratio of approximately 0.2 to approximately 5% based on the total mass of the paste-like dough.

7. The method of claim 5, characterized in that the specific at least one gel-forming hydrocolloid is inulin, and wherein the specific ratio of inulin to the total mass of paste-like dough is approximately 2 to approximately 12%.

8. The method of any preceding claim, characterized in that the pH value of the mixture required for the sterilization of the sprouts / seedlings contaminated with microorganisms is achieved without the addition of additional acidifying agents such as lactic acid, acetic acid, citric acid, or other organic or mineral acids.

9. The method of any preceding claim, characterized in that the inoculum contains an adapted mixed flora of at least one strain of homofermentative and / or heterofermentative lactic acid bacteria, wherein the strain of homofermentative and heterofermentative lactic acid bacteria is selected from the strains L. acidifarinae, L. acidophilus, L. alimentarius, L. amylovorus, L. brevis, L. buchneri, L. cellobiosus, L. coleohominis, L. collinoides, L. crispatus, L. crustorum, L. curvatus, L. delbrueckki, L. diolivorans, L. farciminis, L. fermentum, L. fructivorans, L. frumenti, L. gallinarum, L. gasseri, L. hammesii, L. helveticus, L. hilgardii, L. homohiocchi, L.johnsonii, L. kefiri, L. kimchi, L. kunkeei, L. linderi, L. mali, L. mindensis, L. mucosae, L. ncigelii, L. nantensis, L. namurensis, L. nodensis, L. oris, L. panis, L. porolimentorius, L. parabuchneri, L. porocosei, L. pentosus, L. perolens, L. plantarum, L. pontis, L. reuteri, L. rossiae, L. sakei, L. sanfranciscensis, L. secaliphilus, L. siliginis, L. spicheri, L. vaginalis, L. zymae, including, among others, Lactococcus lactis, Leuconostoc citreum, Lc. argentinum, Lc. suntoryeus, L. gelidum, Lc. mesenteroides, Pediococcus acidilactici, P. damnosus, P. parvulus, P. pentosaceus, Weissella cibaria, Weissella confusa, Weissella kandleri, Weissella paramesenteroides, Weissella viridescens, and mixtures thereof, and optionally there is at least one yeast strain included selected from the strains Candida milleri (Kazachstania milleri), Candida humilis, Kazachstania exigua, Saccharomyces cerevisiae, Debaryomyces hansenii, Dekkera bruxellensis, Kazachstania unispora, Kluyveromyces lactis, S. bayanus, Saccharomyces pastorianus, Torulaspora delbrueckii, T. pretoriensis, Wickerhamomyces anomalus, Pichia anomala, Hansenula anomala, Pichia kudriavzevii, Issatschenkia orientalis, Candida krusei, and mixtures thereof.

10. The method of any preceding claim, characterized in that after the step of providing the mixture of dough with sterilized sprouts / germs, there is a step of further processing the mixture of dough with sterilized sprouts / germs.

11. The method of one of claims 1 to 10, characterized in that after the step of providing the mixture of dough with sterilized sprouts / seedlings, there is a step of isolating the sterilized sprouts / seedlings.

12. The use of a mixture of dough with sterilized sprouts / seedlings produced according to the method of any of claims 1 to 11 for producing baked goods.