WO2022249212A1 - Levain et sa composition pour la préparation de produits alimentaires à base de plantes - Google Patents

Levain et sa composition pour la préparation de produits alimentaires à base de plantes Download PDF

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Publication number
WO2022249212A1
WO2022249212A1 PCT/IT2022/050145 IT2022050145W WO2022249212A1 WO 2022249212 A1 WO2022249212 A1 WO 2022249212A1 IT 2022050145 W IT2022050145 W IT 2022050145W WO 2022249212 A1 WO2022249212 A1 WO 2022249212A1
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WIPO (PCT)
Prior art keywords
sourdough
gluten
mixture
flour
plant
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PCT/IT2022/050145
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English (en)
Inventor
Andrea RAIMONDI
Original Assignee
Felsineoveg S.R.L. - Società Benefit
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Publication of WO2022249212A1 publication Critical patent/WO2022249212A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/045—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with a leaven or a composition containing acidifying bacteria
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D13/00—Finished or partly finished bakery products
    • A21D13/04—Products made from materials other than rye or wheat flour
    • A21D13/045—Products made from materials other than rye or wheat flour from leguminous plants
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D13/00—Finished or partly finished bakery products
    • A21D13/06—Products with modified nutritive value, e.g. with modified starch content
    • A21D13/064—Products with modified nutritive value, e.g. with modified starch content with modified protein content
    • A21D13/066—Gluten-free products

Definitions

  • the present invention relates to a sourdough and a composition thereof for preparing plant-based food products, in particular a sourdough and a composition thereof suitable for decreasing the gluten content in plant-based food products.
  • These foods include naturally gluten-free cereal-based foods such as rice, corn, buckwheat, millet, amaranth, quinoa, cassava, teff, sorghum.
  • the meat substitute food products currently on the market are products based on proteins derived from legumes such as soy or grasses such as wheat.
  • wheat-based meat substitute food products generally consist of a gluten dough.
  • a typical example is toan, a product of oriental origin that is obtained by extracting gluten from wheat flour and then boiling the mixture obtained in water containing soy sauce, kombu seaweed and other flavors.
  • the patent EP 0 821 882 B1 in the name of Green Live Vegetable Meat S.r.l. discloses a method for producing a plant-based meat substitute product consisting of a mixture of gluten and preferably organic, cereal and/or legume flours, which is amalgamated with vegetable broth or water, possibly flavored and spiced, kneaded, molded and then boiled.
  • the final product can be commercialized as a substitute for steaks, stews, sausages, minced meat, hamburgers, ham, bresaola, mortadella, pizzaiola and bacon.
  • the mixture described by the patent consists of a percentage by weight between 45% and 99% of gluten and between 0.1% and 55% of a vegetable flour derived from cereals and/or legumes.
  • the meat substitutes on the market generally have the disadvantage of containing a high percentage by weight of gluten, a protein complex that is hydrolyzed into peptides by the transglutaminase enzyme during digestion in the intestinal tract.
  • a problem encountered in the use of these products concerns people predisposed to celiac disease, who can develop anti-tranglutaminase antibodies causing an inflammatory process that leads the T lymphocytes to "attack" the intestinal villous cells.
  • gliadin a protein present in gluten, activated by the transglutaminase enzyme, activates the T lymphocytes present in the lamina intestinal mucosa, which migrate to the subepithelial site and produce cytokines causing apoptosis of the intestinal villous cells.
  • Patent application WO 2006/097949 A1 in the name of Actial Farmaceutica Lda et Al. describes a sourdough and a leavening composition comprising a mixture composed of at least six species of lactic bacteria and/or Bifidobacteria.
  • the sourdough or leavening composition is used in the preparation of a baked product, which in turn includes the probiotic mixture present in the sourdough or leavening composition.
  • the document also describes the process for preparing the baked product, which includes adding the sourdough or leavening composition to the other ingredients.
  • the aim of the present invention is to solve the aforementioned problems, devising a sourdough and a composition thereof for preparing plant-based food products capable of carrying out proteolytic activity on gluten proteins and decreasing the gluten content of the finished product.
  • a further object of the present invention is a sourdough and a composition thereof for preparing plant-based food products capable of operating synergistically to favor the fermentation processes inside the sourdough, enhancing the proteolytic activity on gluten proteins.
  • a still further object of the present invention is to provide a sourdough and a composition thereof for preparing plant-based food products capable of degrading the allergenic fragments of gluten proteins, achieving a significant qualitative and quantitative reduction of these fragments.
  • a further object of the present invention is to provide a sourdough and a composition thereof for preparing plant-based food products capable of giving the finished food product a taste, consistency and aroma similar to those of meat.
  • a still further object of the present invention is to provide a sourdough and a composition thereof for preparing plant-based food products capable of giving the finished food product a consistency such that the sliceability of the product is improved maintaining the integrity of the product in the packaging step.
  • a further object of the present invention is to provide a sourdough and a composition thereof for preparing plant-based food products capable of improving the shelf life of the food by increasing the shelf-life of the finished product.
  • a further object of the present invention is to provide a plant-based food product with a reduced gluten content.
  • a further object of the invention is to provide a plant-based food product capable of replacing meat and its derivatives, which can be digestible and not harmful to consumers with a predisposition to celiac disease.
  • Another object of the invention is to provide an entirely vegetable-based food product capable of replacing meat and its derivatives while allowing the consumer a varied diet.
  • a still further object of the invention is to provide a plant-based food product containing exclusively plant-based proteins.
  • An even further object of the invention is to provide a plant-based food product with high nutritional properties and of high quality, but relatively low cost.
  • the sourdough for preparing plant-based food products object of the invention is composed of a mixture of flour and water acidified by a complex of lactic bacteria and yeast capable of starting a natural fermentation.
  • the fraction of said lactic bacteria contained within said sourdough is composed of lactic bacteria of the Lactobacillus plantarum species, while the fraction of said yeasts contained within said sourdough consists of yeasts of the Saccharomyces cerevisiae species.
  • the basic dough of said sourdough consists of a mixture of water and flours selected from gluten, cereal and legume flour.
  • the amount of water required to obtain said basic dough of said sourdough varies from 300g to 500g per an overall 1Kg of finished product.
  • said sourdough object of the invention, has shown a particular ability to digest gluten proteins.
  • a composition for formulating said sourdough suitable for decreasing the gluten content in plant-based food products comprises at least one strain of Lactobacillus plantarum and at least one strain of Saccharomyces cerevisiae.
  • Said at least one strain of Lactobacillus plantarum has shown a particular proteolytic capacity of gluten proteins, in particular in the degradation of allergenic fragments of said proteins, leading to a significant qualitative and quantitative reduction of these fragments.
  • the proteolytic activity of said at least one strain of Lactobacillus plantarum is promoted by the presence within the composition of said at least one strain of Saccharomyces cerevisiae, which assists the fermentation processes of the bacterium by hydrolyzing the carbohydrates present in the substrate into simple molecules such as glucose and fructose, more usable by the bacterium, consequently increasing its cellular yield and enhancing its proteolytic activity.
  • Said sourdough is used for formulating and producing plant-based food products, in particular for plant products with a low gluten content.
  • said plant-based food products added with said sourdough show a low gluten content and high digestibility even for those predisposed to celiac disease.
  • said plant-based food products added with said sourdough are composed of gluten in a percentage by weight of from 20% to 40%.
  • said plant-based food products added with said sourdough are composed of at least one type of cereal flour in a percentage by weight from 5% to 15% and/or at least one type of legume flour in a percentage by weight of 10 % to 30%, or with a mixture of at least one type of cereal flour and at least one type of legume flour in a percentage by weight from 10% to 30%.
  • said cereal flours are selected from hard wheat flours and soft wheat flours.
  • Said cereal flours can be bran, wholemeal, fine, extra-fine flours.
  • said cereal flours are organic flours.
  • said legume flours are selected from chickpea, pea, lupine, lentil flours.
  • said legume flours are organic flours.
  • Said mixture of gluten, flours and sourdough is preferably flavored by means of a mixture of spices.
  • said spices present in said mixture are selected from a group comprising pepper, oregano, nutmeg, ginger, chilli and others.
  • Said mixture of cereal and/or legume flours added with said mixture of spices is mixed with water and/or vegetable broth.
  • the quantity of said required water and/or said vegetable broth varies from 200 g to 600 g for each kg of finished product obtained from said mixture of cereal and/or legume flours used.
  • said vegetable oil is selected from extra virgin olive oil and sunflower oil.
  • the process for producing plant-based food products with low gluten content includes the steps of: a) mixing said gluten with at least one said type of cereal flour, at least one said type of legume flour or one said mixture of at least one said type of cereal flour and at least one said type of legume flour; b) adding said mixture obtained with said sourdough; c) flavoring said mixture of gluten, flours and sourdough with a mixture of spices; d) adding water and/or vegetable broth to said mixture and kneading until a homogeneous mixture is obtained; e) stuffing said mixture into a substantially cylindrical or possibly flattened shape, using suitable shapes, with typical sausage casings (fibrous, cellulose, etc.); f) boiling the sausage in water and/or vegetable broth and let it cool. [59]
  • at least one vegetable oil is added to said mixture to give greater elasticity to said mixture.
  • said molded dough is boiled in water and/or vegetable broth for 60-90 minutes and left to cool overnight to obtain a consistency such as to improve the sliceability of the product allowing to maintain the integrity of the product during the packaging step.
  • Figure 1 shows the result of the ITS amplification of the yeasts extracted from the first yeast sample analyzed
  • Figure 2 shows the result of the ITS amplification of the lactic bacteria extracted from the first yeast sample analyzed
  • Figure 3 represents an example of development of the MM6 strain on Gluten Media
  • Figure 4 shows the results of the electrophoretic run of the yeast strains extracted from the second sample of yeast analyzed and of the clone MW1 obtained from the isolation of the first sample of the sourdough;
  • Figure 5 shows the results of the electrophoretic runs of the strains of lactic bacteria extracted from the second sample of yeast analyzed and of the LP1 clone obtained from the isolation of the first sample of the sourdough;
  • Figures 6-10 are chromatograms representing the results of the mass analysis of the sample called "Delicate Yeast” using 70% Ethanol for protein extraction;
  • Figures 11-14 are chromatograms representing the results of the mass analysis of the sample called "Delicate Yeast” using DMF 1.5M for protein extraction;
  • Figures 15, 16 are chromatograms representing the results of the mass analysis of the sample called "Delicate No Yeast” using 70% Ethanol for protein extraction;
  • Figures 17-19 are chromatograms representing the results of the mass analysis of the sample called "Delicate No Yeast” using DMF 1 5M for protein extraction;
  • Figures 20-22 show the separations on gel electrophoresis with optimized conditions of the samples of soft wheat flour, hard wheat flour, gluten, flour mixture, Delicate after TTS and Veghiamo® with the analysis obtained on the densitometer on the side;
  • Figures 23-27 are HPLC chromatograms representing the results of the mass analysis of the samples of gluten extracted with 70% Ethanol, LG4 flour mixture, LG8 flour mixture, Veghiamo® extracted with 70% Ethanol, Veghiamo® after gel extraction electrophoresis;
  • Figures 28-57 are chromatograms representing the results of the mass analysis of the samples of gluten, flour mixture, Delicate after TTS, Veghiamo®.
  • the sourdough for preparing plant-based food products object of the invention is composed of a mixture of flour and water acidified by a complex of lactic bacteria and yeast capable of starting a natural fermentation.
  • the fraction of lactic bacteria contained within the sourdough is composed of lactic bacteria of the Lactobacillus plantarum species, while the fraction of yeast contained within the sourdough consists of yeasts of the Saccharomyces cerevisiae species.
  • the basic dough of the sourdough consists of a mixture of water and flours selected from gluten, cereal and legume flour.
  • the amount of water required to obtain the basic dough of the mother yeast varies from 300g to 500g per overall 1Kg of yeast.
  • the sourdough object of the invention has shown a particular ability to digest gluten proteins.
  • the analysis focused on detecting the presence of lactic bacteria and yeasts, known to be the microorganisms most involved in the fermentation processes useful in the context of food biotransformations.
  • the isolated yeasts all belong to the Saccharomyces cerevisiae species.
  • Lactic acid bacteria were inoculated on Gluten Media, an agar medium containing gluten as the sole source of nitrogen.
  • the six strains analyzed were first developed in MRS culture medium and placed in an incubator at 37°C for 48 hours under anaerobic conditions.
  • the analysis focused on detecting the presence of lactic bacteria and yeasts, known to be the microorganisms most involved in the fermentation processes useful in the context of food biotransformations.
  • Multilocus Sequence Typing was implemented for yeasts, a method aimed at the amplification of microsatellites, while Random Amplification of Polymorphic DNA (RAPD-PCR) was implemented for lactic bacteria.
  • the three isolated clones of the second sample show a profile identical to the clone MW1 purified during the first investigation and are confirmed to belong to the Saccharomyces cerevisiae species.
  • RDM1106 Delicate Yeast, extraction with 70% Ethanol
  • RDM1107 Delicate Yeast, extraction with DMF 1 5M;
  • RDM 1108 Delicate No Yeast, extraction with 70% Ethanol
  • RDM1109 Delicate No Yeast, extraction with DMF 1.5M.
  • P.M. 3909.019 represents the repetitive unit of gliadin-a (33-Mer). Sequence: LQLQPFPQPQLPYPQPQLYPQPQLPYPQPQPF;
  • P.M. 1526.74. 13 particularly immunogenic a-gliadin fragment.
  • Q transformed into E for a total of 4E, P.M. 1530.74.
  • Figures 20-22 show the separations of the samples on gel electrophoresis with optimized conditions with the analysis obtained by the densitometer at the side. The data have been reported through histograms: on the abscissa there are the ranges of molecular weights (PM) in KDa while on the ordinates the average value of the density read by the densitometer.
  • PM molecular weights
  • the areas of interest for the gliadins are between 50-31 KDa. As we can see, the density of gliadins in flour is very high in all the histograms shown.
  • the bands were colored with Coomassie Blue for viewing on the densitometer, the bands were subjected to an initial decolorization step using a 100mM solution of ammonium bicarbonate/acetonitrile (1: 1, V / V), incubated at room temperature and vortexed every 30 minutes. More acetonitrile was added and left to incubate until the bands turned white and folded. The bands were stored at -20 ° C.
  • ⁇ FSQPQQQFPQPQ 12-mer range 1458.678 and fragments.
  • sequences may also be of diagnostic interest:
  • the first analysis concern the extraction of gliadins with EtOH before undergoing separation with gel electrophoresis.
  • the a/b-gliadins elute from 7 min to 16 min.
  • the g-gliadins from 16 min to 18 min.
  • the w-gliadins elute from 18 min to 30 min.
  • LG4, LG8 and “Veghiamo®” are the samples after gel electrophoresis. The presence of high and narrow peaks can be observed due to lipophilic substances remaining in the column.
  • the "Delicate” sample has a greater quantity of toxic fragments than the sample called “Veghiamo®”, but the “Delicate” has only the 10-mer and 12-mer fragments, which are inactive fractions while the “Veghiamo® ’’ contains as inactive fractions the 9-mer, 10-mer and 12-mer fragment of 33-mer.
  • the "Veghiamo®” sample shows a lower quantity of toxic gliadins than the "Delicate after TTS” sample, therefore the fresh yeast is able to further degrade the gluten proteins, resulting in a significant qualitative and quantitative reduction of the allergenic fragments.
  • Figures 28-57 show all the mass analysis indicating the toxic and inactive fragments found.
  • the composition for formulating the sourdough suitable for decreasing the gluten content in plant-based food products object of the present invention comprises at least one strain of Lactobacillus plantarum and at least one strain of Saccharomyces cerevisiae.
  • the at least one strain of Lactobacillus plantarum present in the composition object of the invention has shown a particular proteolytic capacity of gluten proteins, in particular in the degradation of allergenic fragments of said proteins, leading to a significant qualitative and quantitative reduction of these fragments.
  • the proteolytic activity of the at least one strain of Lactobacillus plantarum is promoted by the presence within the composition of the at least one strain of Saccharomyces cerevisiae, which assists the fermentation processes of the bacterium by hydrolyzing the carbohydrates present in the substrate into simple molecules such as glucose and fructose, more usable by the bacterium, consequently increasing its cellular yield and enhancing its proteolytic activity.
  • composition of Lactobacillus plantarum and Saccharomyces cerevisiae for the formulation of a sourdough object of the invention has shown a particular ability to digest gluten proteins.
  • the sourdough containing this composition is used for the formulation and production of plant-based food products, in particular for plant products with a low gluten content.
  • Plant-based food products added with said sourdough show a low gluten content and high digestibility even for those predisposed to celiac disease.
  • the ability of the sourdough to degrade gluten proteins and to reduce allergenic fragments derived from the degradation of these proteins is demonstrated by the aforementioned analysis.
  • the plant-based food products that are added with the sourdough are composed of gluten in a percentage by weight from 20% to 40%, at least one type of cereal flour in a percentage by weight from 5% to 15% and/or at least one type of legume flour in a percentage by weight from 10% to 30%, or with a mixture of at least one type of cereal flour and at least one type of legume flour in a percentage by weight from 10% to 30%.
  • the cereal flours are selected from hard wheat and soft wheat flours.
  • Said cereal flours can be bran, wholemeal, fine, extra-fine flours.
  • said cereal flours are organic flours.
  • the legume flours are selected from chickpea, pea, lupine, lentil flours.
  • said legume flours are organic flours.
  • the mixture of gluten, flours and sourdough is preferably flavored with a mixture of spices.
  • the spices present in said mixture are selected from a group comprising pepper, oregano, nutmeg, ginger, chilli and others.
  • the mixture of cereal and/or legume flours added with the mixture of spices is mixed with water and/or vegetable broth.
  • the quantity of water and/or vegetable broth required varies from 200 g to 500 g for each kg of finished product.
  • the vegetable oil is selected from extra virgin olive oil and sunflower oil.
  • Examples of plant-based food products obtained from the processing of the components described above are the vegetable cold sliced products of the Veghiamo® line.
  • a first example is the lupine vegetable sliced product, which contains water, 25% lupine flour, 22% wheat gluten, sunflower oil, wheat starch, natural flavorings, salt, apple cider vinegar, a mixture of spices and aromatic plants, chickpea flour, durum wheat flour, acidity regulator: sodium bicarbonate, sourdough.
  • a second example is the spiced vegetable sliced product, which contains water, 32% wheat gluten, 8% durum wheat, chickpea flour, extra virgin olive oil, natural flavors, soft wheat flour, salt, a mixture of spices and aromatic plants, sourdough, sunflower oil.
  • a third example is the vegetable sliced product with turmeric and ginger, which contains water, 32% wheat gluten, 9% durum wheat, chickpea flour, sunflower oil, natural flavors, salt, 2% ginger powder, 0.4% turmeric powder, chili powder, sourdough, pepper powder, garlic powder.
  • Gluten is mixed with at least one type of cereal flour, at least one type of legume flour or a mixture of at least one type of cereal flour and at least one type of legume flour.
  • the mixture of gluten, flours and sourdough is flavored with a mixture of spices.
  • the dough is modeled in a substantially cylindrical shape with rounded ends obtaining the typical shape of sausages such as bresaola or salami.
  • the modeled dough is boiled in water and/or vegetable broth for 60-90 minutes and left to cool overnight until a consistency is obtained that improves the sliceability of the product maintaining the integrity of the product during the packaging step.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Botany (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Preparation Of Fruits And Vegetables (AREA)

Abstract

Le levain pour la production de produits alimentaires à base de plantes à faible teneur en gluten comprend une pâte de base comprenant un mélange d'eau et de farine servant de substrat métabolique pour au moins une souche de bactéries lactiques pouvant mettre en œuvre des processus de fermentation et dégrader les protéines gluten et au moins une souche de levure pouvant mettre en œuvre des processus de fermentation et coopérant avec ladite souche de bactéries lactiques. La composition pour la formulation du levain appropriée pour diminuer la teneur en gluten dans des produits alimentaires à base de plantes comprend au moins une souche de Lactobacillus plantarum et au moins une souche de Saccharomyces cerevisiae.
PCT/IT2022/050145 2021-05-24 2022-05-24 Levain et sa composition pour la préparation de produits alimentaires à base de plantes WO2022249212A1 (fr)

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IT102021000013433A IT202100013433A1 (it) 2021-05-24 2021-05-24 Lievito madre per la preparazione di prodotti alimentari a base vegetale
IT102021000013433 2021-05-24

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006097949A1 (fr) * 2005-03-16 2006-09-21 Actial Farmacêutica, Lda. Melange d’au moins 6 especes de bacteries de l'acide lactique et/ou bifidobacteria dans la facrication du levain
WO2010073283A2 (fr) * 2008-12-23 2010-07-01 Giuliani S.P.A. Procédé de biotechnologie biomicrobienne pour dégrader complètement le gluten dans des farines
DE202016101614U1 (de) * 2016-03-23 2016-05-02 Ernst Böcker Gmbh & Co. Kg Glutenfreie Backwaren

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT1284198B1 (it) 1996-07-30 1998-05-08 Beniamino Anzalone Carne vegetale

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006097949A1 (fr) * 2005-03-16 2006-09-21 Actial Farmacêutica, Lda. Melange d’au moins 6 especes de bacteries de l'acide lactique et/ou bifidobacteria dans la facrication du levain
WO2010073283A2 (fr) * 2008-12-23 2010-07-01 Giuliani S.P.A. Procédé de biotechnologie biomicrobienne pour dégrader complètement le gluten dans des farines
DE202016101614U1 (de) * 2016-03-23 2016-05-02 Ernst Böcker Gmbh & Co. Kg Glutenfreie Backwaren

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