Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
  • A23L7/10—Cereal-derived products
  • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
  • A23L29/065—Microorganisms
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system

Definitions

• the present invention relates to a consumable product containing probiotics and to a process for obtaining it .
• Probiotic micro-organisms are micro-organisms which beneficially affect a host by improving its intestinal microbial balance. In general, it is believed that these bacteria inhibit or influence the growth and/or metabolism of pathogenic bacteria in the intestinal tract . It is also assumed that via probiotic micro-organisms the immune function of the host is activated. For this reason, there have been many different approaches to include probiotic micro-organisms into food-stuffs.
• WO98/10666 discloses a process for manufacturing a dehydrated food composition containing live probiotic acid bacteria, in which a food composition and a culture of probiotic lactic acid bacteria sensitive to oxygen are sprayed conjointly under a stream of hot air.
• EP0862863 discloses a dried, ready-to-eat cereal product comprising a gelatinised starch matrix which includes a coating or filling containing a probiotic microorganism.
• US4943437 discloses a process for supply of biologically active materials to base food materials, in which th biologically active material is slurried in an inert carrier, where it is insoluble, to form a homogeneous suspension, after which the suspension is applied to the base material .
• GB2205476 (UNILEVER) discloses a supported bacterial composition comprising an inert subdivided support, which is flour, and an aqueous suspension of viable microflora. This mixture is then dried and is suitable as inoculum of lactic acid bacteria for the preparation of sour-dough bread.
• probiotics probiotic micro-organisms
• One first goal to reach is to have an adequate number of cfu (colony forming unit) per day. If the concentration of the probiotics in the product does not exceed a certain threshold value, the beneficial effect is not provided.
• an effective dose is in the range of 10 9 cfu per human per day, and, supposing, that the consumer has to take them within his/her daily intake, it is the objective to deliver this amount of cfu within one to three servings.
• the approach has been to use probiotics that have been dried, either per se or together with a supporting substance.
• the probiotics are usually concentrated, for example by centrifugation or filtration, and are then dried by spray- drying, fluidized-bed drying or freeze-drying. From the drying process, another, serious problem arises. That is, the probiotics sustain substantial loss in the range of 60, more frequently 90 to 99 % of cfu depending on the applied technology, unless special measurements are taken. It goes without saying that these drying steps are very energy- expensive. But the high temperature drying process has other disadvantages. It may destroy or impair metabolites that are present in the probiotics them-selves or in the fermented medium where they were cultivated. Such metabolites may therefore lose their beneficial effects.
• the disadvantage of a concentration step is the loss of metabolites that were present in the fermented medium.
• the powder obtained by drying may then be applied to the desired food-product .
• the dried probiotics are mixed with a liquid carrier substance, which is either oil, water or a protein digest. Then this substance is sprayed onto the food-product .
• probiotics Due to the need of a relatively high number of cfu within a single meal and the high losses during drying, it is a problem to have a food-product with an effective number of cfu.
• Another concern is the viability of the probiotics in the stomach and the intestine.
• the probiotics must be sufficiently resistant to the acid environment in the stomach and to the bile acids in order to successfully colonize the intestine.
• the food-product comprising probiotics must be palatable to the consumer. There is a need to apply probiotics to a food-product without notably influencing its organoleptic properties.
• the present invention provides a consumable product comprising probiotics, wherein the probiotics were freshly applied to it.
• the present invention provides a consumable product comprising metabolites produced by probiotics wherein the metabolites were comprised in a fermented medium that was separated from the probiotics cultivated therein.
• the process for obtaining a consumable product comprising probiotics comprises producing a fresh biomass of probiotics by fermentation in a liquid medium and directly applying the fresh biomass to the consumable product .
• the process for obtaining a consumable product comprising metabolites produced by probiotics comprises cultivating probiotics in a liquid medium, separating the liquid medium from the probiotics and directly applying the liquid medium to the consumable product.
• biomass derived from a fermentation process can be directly and freshly applied to a consumable product without high temperature drying.
• a consumable product containing probiotics is obtained, which has an excellent storage stability and which has an appearance and organoleptic properties similar to the appearance and the organoleptic properties of a similar consumable product not containing probiotics.
• the consumable product if consumed in the expected or reasonable amount, contains an amount of cfu that is sufficient to exert a beneficial effect.
• metabolites and micro-organisms are no longer lost due to drying process and concentration.
• consumer product means a product which is consumable by humans and/or by pets such as dogs or cats, for example.
• fresh probiotics or “freshly applied biomass” refers to probiotics that, after the fermentation process, are not dried, for example by spray-, fluidized bed or freeze-drying.
• fresh probiotics is not intended to be understood as biomass that is applied within a certain time limit to the consumable product. It is easily possible to store the "fresh biomass” for a certain time without loss. If the biomass can also be frozen for a certain time and thawed out without substantial loss, this is still considered as fresh.
• probiotics for the purpose of the present invention, the term "probiotics", “probiotic micro-organism” or probiotic biomass is understood to include any micro-organisms, cell content or metabolites from micro-organisms, having beneficial effects to its host. Therefore, yeasts, moulds and bacteria may be included.
• EP 0862863 lists some examples for probiotics presently known. For example, strains of Lactobacillus j ohnsonii (CNCM 1-1225) , Bifidobacterium lactis (DSM20215), Streptococcus thermophilus (TH4, Chr. Hansen, DK) , or Lactobacillus paracasei (CNCM 1-2116) may be used.
• a selection of different probiotic strains is offered by Christian Hansen BioSystems A/S (CHL) , 10-12 Boge Alle, P.O Box 407, DK-2970 Horsholm, Denmark.
• probiotics furthermore is intended to include the metabolites generated by the micro-organisms during a fermentation process, if they are not separately indicated. These metabolites may be released to the medium of fermentation or they may be stored within the microorganism. It may well be, that such metabolites are responsible for part or all of the beneficial effects of a particular probiotic micro-organism.
• probiotics need not necessarily be concentrated and don't need to be dried at high temperatures but can be directly and freshly applied to a food product.
• the present invention has therefore also the big advantage, that there isn't a high temperature treatment that may impair or even destroy the effectivity of metabolites produced by the probiotics.
• concentration step can be omitted has the advantage that effective metabolites present in the fermented medium are not lost, for example by filtration. CQ 1 >.
• probiotics in high abundance to the consumable product.
• a relatively small amount of slurry or liquid from the fermentor comprising probiotics has to be applied to the consumable product .
• a comparatively slight abundance of probiotics still may be applied to the consumable product in order to compensate for the inevitable losses during storage as well as passage through the digestive tract of the product .
• probiotics applied to a food product show, depending on the species and strain of the probiotic organism, sufficient resistance to the environment of the stomach and the gastric and bile acids ⁇ in vi tro tests) .
• At least one protective agent may be added to the probiotics prior to their application to the consumable product, for example.
• the probiotics according to the invention may be obtained by fermentation and they may be stored after fermentation and before application to the consumable product for a time and at a temperature that prevents substantial loss of probiotic cfu, for example. It is clear that the biomass, after termination of the fermentation or cultivation, may be stored for a certain time. In experiments, the biomass of different probiotics was stored for 4 days at 5°C without detectable loss. Furthermore, also the resistance to gastric or bile acid ⁇ in vi tro tests) was not influenced by storage time.
• the probiotics may be fermented until a final concentration of 10 6 to 5 xlO 10 , preferably 10 7 to 3 xlO 10 , more preferably 1.5 xlO 7 to 10 10 , even more preferably 10 8 to 9.5 xlO 9 , in particular 2 to 9 xlO 9 cfu per ml of fermented medium is achieved, for example .
• the probiotics to be applied to the food product are concentrated to a final concentration of 10 7 to 10 12 , preferably 10 8 to 5 xlO 11 , more preferably 1.5 xlO 8 to 10 11 , even more preferably 10 9 to 5 xlO 10 cfu per ml of fermented medium, for example .
• any probiotic micro-organism may be used.
• a probiotic strain or strains may be selected from a group comprising yeasts, preferably the genus Saccharomyces, moulds, preferably the genus Aspergillus, bacteria, preferably the genus Lactobacillus , Bifidobacterium, Streptococcus , Enterococcus, and a mixture thereof.
• yeasts preferably the genus Saccharomyces, moulds, preferably the genus Aspergillus
• bacteria preferably the genus Lactobacillus , Bifidobacterium, Streptococcus , Enterococcus, and a mixture thereof.
• Lactobacillus johnsonii Bifidobacterium lactis, Streptococcus thermophilus, or, Lactobacillus paracasei may be used.
• strains may be selected from the geni Lactobacillus , Streptococcus , Bifidobacterium, Bacteroides , Clostridium, Fusobacterium, Melissococcus , Propionibacterium, Enterococcus, Lactococcus, Staphylococcus , Peptostreptococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus .
• a probiotic strain or strains may be selected from a group comprising Bifidobacterium lactis (DSM20215) , Lactobacillus johnsonii (1-1225 CNCM) , Lactobacillus paracasei (1-2116 CNCM) , Streptococcus thermophilus (TH4 , Chr. Hansen, DK) , mixtures thereof, and a mixture also comprising other probiotic micro-organisms, for example.
• the percentage of fresh biomass of probiotics added to the consumable product may be 0.05 to 4%, preferably 0.1 to 1.5%, most preferably 0.2 to 1% by weight of the consumable product, for example.
• the final concentration of the probiotics applied to the consumable product may be 10 s to 10 9 , more preferably, 10 7 to 10 8 , most preferably 2 to 8 xlO 7 cfu/g with respect to the total weight of the consumable product, for example .
• the fermented medium may have been directly applied to the consumable product .
• the fermentation may be kept ongoing until a final concentration of 10 s to 5 xlO 10 , preferably 10 7 to 3 xlO 10 , more preferably 1.5 xlO 7 to 10 10 , even more preferably 10 8 to 9.5 xlO 9 , in particular 2 to 9 xlO 9 probiotic cfu per ml of fermented medium is achieved, for example.
• the process of the present invention may comprise, before applying the fresh biomass to a consumable product, concentrating the biomass to a final concentration of 10 7 to 10 12 , preferably 10 8 to 5 xlO 11 , more preferably 1.5 xlO 8 to 10 11 , even more preferably 10 9 to 5 xlO 10 cfu per ml of fermented medium, for example.
• the Aw of the consumable product at at the beginning and/or during shelf life is below 0.5.
• the Aw of the consumable product is below 0.4 and more preferably it is smaller than 0.3.
• the Aw of the consumable product is below 0.2.
• the Aw is in the range of 0.005 to 0.3, or 0.01 to 0.15 during the shelf life of the consumable product.
• the consumable product has a packaging that substantially limits the water uptake from the environment .
• the 0 2 permeation rate of the packaging of the consumable product is preferably below 4.2ml/m 2 d, preferably below 3.8ml/m 2 d.
• the water vapor transmission rate (WVTR) of the packaging of the consumable product is preferably below 3.5g/m 2 d, more preferably below 3 g/m 2 d.
• the packaging may comprise co-extruded cross-linked oriented low density polyethylene (LDPE) .
• the bags may be hermetically sealed, for example heat-sealed.
• the purpose of the packaging as characterized above is to maintain the preferred Aw values during the shelf life of the consumable product.
• the shelf life of the product may be up to 6 months, preferably up to 12 months, more preferably up to 18 months and most preferably up to two years .
• the process may further comprise, after fermentation, storing the fresh biomass for a time and at a temperature that prevents substantial loss of probiotic cfu, for example.
• the process may further comprise, before, during or after producing fresh biomass of probiotics, adding of at least one protective agent to the medium of fermentation or to the fresh probiotic biomass, for example.
• the fermentation process according to the present invention may be kept ongoing for 6 hours to 3 days, preferably 6 to 20 hours, more preferably 7 to 17 hours, depending on the strain of probiotic micro-organism used, for example.
• the same strain or strains may be used as described above with respect to a consumable product comprising probiotics, for example.
• the percentage of fresh biomass of probiotics added to the consumable product may be 0.05 to 4%, preferably 0.1 to 1.5%, most preferably 0.2 to 1% by weight of the consumable product, for example.
• the final concentration of the probiotics applied to the consumable product may be 10 s to 10 9 , more preferably, 10 7 to 10 8 , most preferably 2 to 8 xlO 7 , in particular 5 xlO 7 cfu/g of the consumable product.
• the biomass freshly derived from the fermenting process be concentrated.
• concentration can be achieved by centrifugation or filtration.
• the level of concentration allows dosing accurately the amount of cfu per gram of consumable products.
• the concentration may also take into account the subsequent loss of cfu during shel - life of the food-product or during passage of the digestive tract.
• a high-temperature drying process can be avoided by spraying or otherwise applying not concentrated or- relatively little concentrated biomass to the consumable product, so that the water activity of the overall product does not decisively increase.
• a high temperature drying process is not necessary due to "absorbtive drying"; the already dried food product absorbs rapidly the water accompanied by and contained in the probiotic biomass. The exposure to room temperature during the process of application is sufficient to prevent a decisive increase of water activity of the final product.
• the supernatant obtained thereby need not be discarded.
• the medium after the fermentation with probiotics usually contains metabolites having similar beneficial effects as the probiotics them-selves. Therefore, the supernatant medium may, after concentration of the biomass, also be applied to a consumable product .
• all kind of starting consumable products may be used.
• Food and beverages for humans as well as pet food may be enriched by probiotics.
• nutritional formulas for each and every purpose may be supplied with probiotics.
• nutritional formulas for example for sportsmen or athletes, for people with special nutritional needs such as people allergic to certain natural food components or people with gastrointestinal disorders and so forth.
• chocolate or other sweet products may be supplied with probiotics.
• all kind of extruded or cooked or otherwise prepared food products may be furnished with probiotics.
• dried products may be used, such as dried pet food or other dried food products, like for example powders, flours, milk or cereal powders or cereal flakes.
• Probiotics may be used to be applied to all kind of ⁇ . ⁇ 4-1
• strains used for the examples are the following:
• Table 1 Composition and production of consumable products referred to in the examples
• Example 1 Bifidobacterium lactis biomass applied to different products :
• Bifidobacterium lactis was fermented and then concentrated by centrifugation. Details of the fermentation are given in tables 2 and 3 below. Standard protective agents were added to the concentrate. This biomass was added in bench-scale to different commercial available cereal products (see table 1 above) .
• Table 2 Medium composition for Bifidobacterium lactis (example 1) .
• Example 2 Bifidobacterium lactis , Lactobacillus johnsonii , Lactobacillus paracasei . Streptococcus thermophilus biomass applied to a junior cereal product
• Table 7 Medium composition for Lactobacillus johnsonii (example 2 ) .
• Table 9 Medium composition for Streptococcus thermophilus (example 2) .
• Table 11 Medium composition for Lactobacillus paracasei (example 2) .
• Table 13 Results of application trials on a junior cereal product .
• Example 3 Shelf life data on a junior cereal product with Lactobacillus johnsonii
• Lactobacillus johnsonii was fermented and then concentrated by centrifugation (for fermentation details see tables 14 and 15) . Standard protective agents were added to the concentrate. This biomass was added in pilot-scale to a junior cereal product.
• Table 14 Medium composition for Lactobacillus johnsonii (example 3 )
• Table 16 Results of application and shelf life on a junior cereal product .
• the shelf-life study reveals that storage for up to one year does not substantially reduce the number of cfu on the product .
• Example 4 Addition of concentrated and non-concentrated Bifidobacterium lactis , direct and after 4 days storage of biomass to a junior cereal product
• Bifidobacterium lactis was fermented (details are given in tables 17 and 18) , a part of the biomass was used directly and a second part was concentrated by centrifugation with addition of standard protective agents. Both biomasses were added bench-scale to a junior cereal product. A second series of trials was conducted with the same biomasses stored at 5°C for 4 days prior to application.
• For the bench-scale application 2 kg of cereal product was put into a rotating batch coating drum and the biomass was sprayed on top of the rotating cereals using a commercial spray pistol with a two-phase (air/liquid) nozzle. In all cases 0.5% of the total cereal amount was added.
• the biomass was used directly after fermentation and in the other case it was concentrated and then the same steps were repeated with biomasses stored for 4 days (5°C) prior to application.
• the finished products were also analysed In vi tro for gastric tract resistance.
• Table 17 Medium composition for Bifidobacterium lactis (example 4 ) .
• Table 19 Results of application trials on a junior cereal product .

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• 2002
  • 2002-02-12 US US10/468,645 patent/US8263146B2/en not_active Expired - Fee Related
  • 2002-02-12 RU RU2003128070/13A patent/RU2302747C2/ru not_active IP Right Cessation
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  • 2002-02-12 EP EP02726108A patent/EP1408760B1/en not_active Revoked
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