Compositions and methods for producing fermented pea proteins

Abstract

Described herein are compositions and methods for producing a pea protein ferment. A starting composition comprising pea protein, a Leuconostoc citreum or Leuconostoc pseudomesenteroides bacterium and sucrose can be fermented for a period of time under conditions suitable to produce a pea protein ferment. The resulting pea protein leavening will have increased viscosity, improved appearance and improved sensory properties relative to an equivalent pea protein composition that has not been contacted with or fermented by the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of European Patent Application No. 22206070.9, filed on November 8, 2022, which is hereby incorporated by reference in its entirety. Background of the Invention

[0003] With the increasing public interest in plant - based proteins, pea protein is finding wider and wider applications in the food and beverage industries. For example, pea protein can be found in many commercially available energy bars, meal replacement shakes, plant - based meat alternatives, breakfast cereal products, supplement products, etc. Despite the many commercially available sources of pea protein and the many commercially available pea - protein - containing products, there is an opportunity to improve not only the flavor and sensory properties of pea protein but also its functional properties.

[0004] Fermentation is an ancient and widely used process for altering the flavor and functional properties of foods. For example, the fermentation of cabbage can produce sauerkraut and kimchi products, the fermentation of milk can produce cheese and yogurt, and the fermentation of fruits, sugars, and grains can produce alcoholic beverages. However, the practice of fermentation still has many widespread applications and potential yet to be discovered and developed.

[0005] Compositions and methods are described herein for fermenting pea protein, resulting in beneficial improvements in both sensory appearance and functional characteristics. Summary of the Invention

[0006] The present disclosure provides a composition containing pea protein, bacteria of Leuconostoc citreum or Leuconostoc pseudomesenteroides, and sucrose. The composition may comprise pea protein between 1 wt% and 20 wt%, between 2 wt% and 18 wt%, or between 4 wt% and 15 wt%, and / or sucrose between 1 wt% and 30 wt%, between 2 wt% and 25 wt%, or between 5 wt% and 20 wt%. The bacteria of Leuconostoc citreum or Leuconostoc pseudomesenteroides are optionally selected from the group consisting of: Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number LMG P-32799), and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195). The composition may be fermented for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; and / or the composition may be fermented at a temperature between 20 °C and 30 °C, between 22 °C and 28 °C, between 24 °C and 26 °C, or about 25 °C.

[0007] The present disclosure also provides a method for fermenting pea protein, the method comprising: (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for a period of time under conditions sufficient to produce a fermented pea protein product. The pea protein composition may comprise between 1 wt% and 20 wt%, between 2 wt% and 18 wt%, or between 4 wt% and 15 wt% pea protein, and / or between 1 wt% and 30 wt%, between 2 wt% and 25 wt%, or between 5 wt% and 20 wt% sucrose. The Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria may be selected from the group consisting of: Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number LMG P-32799), and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195). The composition may be fermented for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; and / or the composition may be fermented at a temperature between 20°C and 30°C, between 22°C and 28°C, between 24°C and 26°C, or at about 25°C. The method may further comprise a step of stirring the fermented pea protein product during or after step (i). The method may be a method for increasing the viscosity of pea protein, and the fermented pea protein product has a higher viscosity compared to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria; and / or the method may be a method for changing the appearance of pea protein, and the fermented pea protein product has a glossier, shinier, stickier, and / or more gel-like appearance compared to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria; and / or the method may be a method for changing one or more sensory properties of the pea protein composition, and the fermented pea protein product has an increased sour taste, a reduced green pea flavor, an increased sweet taste, and / or a reduced grass flavor compared to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria.

[0008] The present disclosure also provides a composition comprising a fermented pea protein product obtained by the method described herein. The composition may comprise fructose, α-glucan, polyols, and / or organic acids, and optionally, wherein the composition does not contain sucrose and / or wherein the composition does not contain starch. The composition may comprise between 1 wt% and 20 wt%, between 2 wt% and 15 wt%, or between 5 wt% and 10 wt% α-glucan.

[0009] The present disclosure also provides the use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for increasing the viscosity of pea protein by fermenting the pea protein with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose.

[0010] The present disclosure also provides a method for obtaining a shiny pea protein product, the method comprising: (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria at a temperature between 20 °C and 30 °C, between 22 °C and 28 °C, between 24 °C and 26 °C, or about 25 °C for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; (ii) stirring the contacted pea protein composition during or after step (i); and (iii) obtaining the stirred fermented pea protein product having a shinier appearance than the pea protein composition before contact with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria.

[0011] The present disclosure also provides isolated Leuconostoc citreum or Leuconostoc pseudomesenteroides bacterial cells selected from the group consisting of: Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number LMG P-32799), and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195). BRIEF DESCRIPTION OF THE DRAWINGS

[0012] This patent or application contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Patent Office upon request and payment of the necessary fee.

[0013] The drawings generally illustrate, by way of example, and not by way of limitation, the various aspects described herein.

[0014] Figure 1 Photographs showing the visual appearance of fermented samples 1.1 - 1.8 before and after stirring, as outlined in Example 1.

[0015] Figure 2 Photographs showing the visual appearance of fermented samples 1.9 - 1.16 before and after stirring, as outlined in Example 1.

[0016] Figure 3 Photographs showing the visual appearance of fermented samples 1.17 - 1.24 before and after stirring, as outlined in Example 1.

[0017] Figure 4 Still frames from a video showing the appearance of comparative samples 1.17 and 1.24.

[0018] Figure 5 Shows the viscosity curves of the fermentates of Leuconostoc citreum B3K7 for samples 1.1 - 1.8.

[0019] Figure 6 Shows the viscosity curves of the fermentates of Leuconostoc citreum C22B11 for samples 1.9 - 1.16.

[0020] Figure 7 Shows the viscosity curves of the fermentates of Leuconostoc pseudomesenteroides C18X24 for samples 1.17 - 1.24.

[0021] Figure 8 Shows the reduction in syneresis for sample 1.2 relative to the control starch and protein suspension samples.

[0022] Figure 9 Shows a comparison of the appearance between the reference vegan dessert formulation made with pea protein, starch, and carrageenan and the appearance of samples 1.2 and 1.10.

[0023] Figure 10 Shows the appearance and consistency of the reference vegan dessert formulation made with pea protein, starch, and carrageenan compared to samples 1.2, 1.10, and 1.18.

[0024] Figure 11 Shows the relative changes in sensory attributes when sample 1.2 is compared to the reference vegan dessert formulation outlined in Example 4.

[0025] Figure 12 Shows the viscosity of the samples outlined in Example 5.

[0026] Figure 13 Shows the viscosity curves of the samples of Scheme A from Example 5.

[0027] Figure 14 Shows the viscosity curves of the samples of Scheme B from Example 5.

[0028] Figure 15 Shows the viscosity curves of the samples of Scheme C from Example 5.

[0029] Figure 16 Shows the residual sucrose concentration of the samples outlined in Example 5.

[0030] Figure 17 Shows the viscosity of the samples outlined in Example 6.

[0031] Figure 18 Shows the viscosity curves of the 4 wt% pea protein fermentation samples from Example 6.

[0032] Figure 19 Shows the viscosity curve of an 8 wt% pea protein fermentation sample from Example 6.

[0033] Figure 20 Shows the viscosity curve of an active wheat gluten fermentation sample from Example 6.

[0034] Figure 21 Shows the viscosity curve of a zein fermentation sample from Example 6. Detailed Description

[0035] Reference will now be made specifically to certain aspects of the subject matter disclosed in the present invention, examples of which are shown, in part, in the accompanying drawings. While the disclosed subject matter will be described in connection with the enumerated claims, it should be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0036] In this document, unless the context clearly dictates otherwise, the terms "a," "an," or "the" are used to include one or more than one. Unless otherwise indicated, the term "or" is used to mean non-exclusive "or." All publications, patents, and patent documents cited in this document are hereby incorporated by reference in their entirety as if each were incorporated by reference individually. If there is an inconsistency in the usage between this document and those documents incorporated by reference in this manner, the usage in the incorporated references shall be regarded as a supplement to the usage in this document; for irreconcilable inconsistencies, the usage in this document shall prevail.

[0037] Values expressed in a range format should be interpreted in a flexible manner to include not only the explicitly recited values that are the limits of the range, but also all individual values or sub-ranges subsumed within that range as if each value and sub-range were explicitly recited. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not only about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the specified range. Unless otherwise indicated, the statement "about X to Y" has the same meaning as "about X to about Y." Similarly, unless otherwise indicated, the statement "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z."

[0038] Unless explicitly specified, ppm (parts per million), percentages, and ratios are based on weight. Percentages by weight are also referred to hereinafter as wt% or (wt)%.

[0039] The present disclosure relates to compositions and methods for producing fermented pea protein. As described herein, fermented pea protein products are characterized by increased viscosity, altered visual appearance, and / or altered one or more sensory attributes relative to the protein prior to fermentation. Generally, pea protein is fermented with Leuconostoc citreum bacteria.

[0040] Fermentate

[0041] The present disclosure relates to compositions comprising pea protein, Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria, and sucrose, and methods of using the compositions to produce fermented pea protein products.

[0042] Generally, the starting composition will include pea protein (i.e., protein extracted and / or derived from the seeds or pod fruit of Pisum sativum). Pea protein can be from any suitable source. Pea protein can be pea protein isolate, pea protein concentrate, or a combination thereof. Suitable pea proteins are commercially available and can include, but are not limited to (Roquette), Pea Protein (COSUCRA TM ), RADIPURE TM Pea Protein Isolate and Pea Protein 870 Suitable pea protein compositions can contain at least 50%, at least 60%, at least 70%, at least 75%, or at least 80% protein. The starting composition can contain between 1 wt% and 20 wt%, between 2 wt% and 18 wt%, or between 4 wt% and 15 wt% pea protein. For example, the starting composition can contain equal to or about 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, or 20 wt% pea protein.

[0043] The starting composition further comprises sucrose. The composition may comprise sucrose in an amount between 1 wt% and 30 wt%, between 2 wt% and 25 wt%, or between 5 wt% and 20 wt%, for example, equal to or about 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25 wt%, 26 wt%, 27 wt%, 28 wt%, 29 wt% or 30 wt% of sucrose. The sucrose can be from any suitable source. Those skilled in the art will recognize suitable sources, including commercial sources or sucrose.

[0044] The starting composition may comprise pea protein and sucrose in a weight ratio between about 3:1 to 1:10, between about 2:1 to 1:8, or between about 1:1 to 1:4, preferably between about 1:1 to 1:4.

[0045] The starting composition further comprises Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria. The Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria can be from any suitable source. Suitable Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria include but are not limited to Leuconostoc citreum strain B3K7 (deposited at the BCCM / LMG Bacteria Collection, Ghent University, K.L. Ledeganckstraat 35, 9000 Gent, Belgium on September 27, 2022, accession number LMG P-32801), Leuconostoc citreum strain C22B11 (deposited at BCCM / LMG, Ghent University, K.L. Ledeganckstraat 35, 9000 Gent, Belgium on September 27, 2022, accession number LMG P-32800), Leuconostoc pseudomesenteroides strain C18X24 (deposited at BCCM / LMG, Ghent University, K.L. Ledeganckstraat 35, 9000 Gent, Belgium on June 21, 2023, accession number LMG P-33195), Leuconostoc citreum strain C18X1 (deposited at BCCM / LMG, Ghent University, K.L. Ledeganckstraat 35, 9000 Gent, Belgium on October 20, 2022, accession number LMG P-32799) and combinations thereof.

[0046] In the priority application EP 22206070.9 filed on November 8, 2022, the strain C18X24 was incorrectly represented as strain C18X1 in the examples and throughout the specification, drawings, and claims. Appropriate corrections are made herein. Accordingly, data using strain C18X24 is provided in application EP 22206070.9, and strain C18X24 was fully supported as of the filing date of the priority application (November 8, 2022). Additional data is provided herein based on strain C18X1, and this strain is different from the incorrectly labeled strain in the priority application.

[0047] The starting composition is fermented for a period of time under conditions sufficient to produce a pea protein fermentate. For example, the pea protein can be contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose at a temperature between 20 °C and 30 °C, between 22 °C and 28 °C, between 24 °C and 26 °C, or at about 25 °C for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours.

[0048] As used herein, "fermented pea protein product" and "pea protein fermentate" are used interchangeably and refer to a composition produced by microbial fermentation of pea protein and include (i) said pea protein; (ii) metabolites produced by the microorganism during pea protein fermentation; (iii) non-living microorganisms used in the fermentation process; and (iv) water. The pea protein fermentate can include metabolites such as, but not limited to, fructose, α-glucan, polyols, organic acids, and combinations thereof. For example, the pea protein fermentate can include between 1 wt% and 20 wt%, between 2 wt% and 15 wt%, or between 5 wt% and 10 wt% α-glucan. The pea protein fermentate can include between 1 wt% and 20 wt%, between 2 wt% and 15 wt%, or between 5 wt% and 10 wt% fructose. The pea protein fermentate can include between 0.1 wt% and 10 wt%, between 0.5% and 8%, or between 1 wt% and 5 wt% polyols. The pea protein fermentate can include between 0.1 wt% and 10 wt%, between 0.5% and 8%, or between 1 wt% and 5 wt% organic acids. The pea protein fermentate can include between 0.01 wt% and 5 wt%, between 0.05 wt% and 2 wt%, or between 0.1 wt% and 1 wt% dietary fiber. In one example, the pea protein fermentate can include between 5 wt% and 10 wt% fructose, between 5 wt% and 10 wt% α-glucan, between 1 wt% and 5 wt% polyols, between 1 wt% and 5 wt% organic acids, between 0.1 wt% and 1 wt% dietary fiber, protein, fat, and water.

[0049] The pea protein fermentate may include α-glucan, which is a linear α-glucan. Generally, the α-glucan may have an average molecular weight of at least 300 kDa, at least 500 kDa, at least 750 kDa, at least 1 MDa, at least 2 MDa, at least 3 MDa, at least 4 MDa, at least 5 MDa, at least 6 MDa, at least 7 MDa, at least 8 MDa, or about 9 MDa. The α-glucan may have an average molecular weight between 300 kDa and 9 MDa.

[0050] The pea protein fermentate can be processed using an inactivation step in which the microorganisms are rendered non-viable. For example, the pea protein fermentate can be pasteurized, heat-inactivated, irradiated, or chemically treated to render any remaining microorganisms non-viable. The pea protein fermentate can additionally or alternatively undergo a physical method by which the microorganisms are separated, such as by filtration.

[0051] Although sucrose is used in the production of the pea protein fermentate, the resulting pea protein fermentate may be free of sucrose. In other words, all of the sucrose present in the initial starting composition can be utilized by Leuconostoc citreum or Leuconostoc pseudomesenteroides during fermentation such that the resulting pea protein fermentate is free of sucrose. In one example, the pea protein fermentate can include between 5 wt% and 10 wt% fructose, between 5 wt% and 10 wt% α-glucan, between 1 wt% and 5 wt% polyols, between 1 wt% and 5 wt% organic acids, between 0.1 wt% and 1 wt% dietary fiber, protein, fat, and water, and be free of sucrose (e.g., less than 1 wt%, less than 0.5 wt%, less than 0.1 wt%, less than 0.01 wt%, or less than the detection level of sucrose). Similarly, the pea protein fermentate can be free of added sucrose, whereby all of the sucrose in the starting composition is depleted and no additional sucrose is added to the resulting pea protein fermentate.

[0052] The pea protein fermentate can be free of added starch. As used herein, "free of added starch" means a composition in which no starch component has been added, but may include starch produced as a result of the fermentation process or reaction. For example, the pea protein fermentate can include starch produced by microorganisms during fermentation, but no other starch component is added.

[0053] Additionally, the pea protein fermentate can be stirred to form a stirred pea protein fermentate. The pea protein fermentate can be stirred manually or mechanically. The pea protein fermentate can be stirred for at least 2 seconds, 5 seconds, 10 seconds, 15 seconds, 30 seconds, 45 seconds, or 60 seconds, and / or until a desired texture is obtained.

[0054] The pea protein fermentate can have a pH between about 4 and 5, between 4.1 and 4.8, or between 4.2 and 4.7. Generally, a higher concentration of sucrose in the starting composition will result in a pea protein fermentate with a slightly higher pH value.

[0055] Generally, the pea protein fermentates described herein are characterized by an increase in viscosity relative to equivalent pea protein compositions that have not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria / have not been fermented with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria. When measured after stirring at 250 rpm and 25 °C for 5 minutes, the viscosity of the pea protein fermentate can be at least 700 cP, 800 cP, 900 cP, 1000 cP, 1100 cP, 1200 cP, 1300 cP, 1400 cP, 1500 cP, 1800 cP, 2000 cP, 2200 cP, 2500 cP, 2800 cP, 3000 cP, 3200 cP, 3500 cP, 3800 cP, or at least 4000 cP. Generally, a higher concentration of pea protein in the starting composition will result in a pea protein fermentate with a higher viscosity. Similarly, a higher concentration of sucrose in the starting composition will result in a pea protein fermentate with a higher viscosity. Thus, as is evident from the data provided herein, one of ordinary skill in the art can customize the starting composition to vary both the pea protein and sucrose concentrations to produce a pea protein fermentate with a particular desired viscosity.

[0056] The pea protein fermentates described herein are characterized by a change in visual and physical appearance relative to equivalent pea protein compositions that have not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria / have not been fermented with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria. For example, compared to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria / has not been fermented with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria, the pea protein fermentate can be glossier, shinier, more viscous, and / or have a more gel-like appearance. A "glossy" appearance of the pea protein fermentate is an appearance in which the pea protein fermentate is smooth (i.e., free of lumps or granules in appearance) and shiny. A "shiny" appearance is an appearance in which the surface of the pea protein fermentate reflects light. A "viscous" appearance is an appearance in which the pea protein fermentate appears to be soft and sticky. A "gel-like" appearance is an appearance in which the pea protein fermentate appears to be thick, slightly sticky, and somewhat firm / solid. The evaluation of the appearance of the pea protein fermentate can be made with the naked eye or can be assisted by mechanically measuring one of the above characteristics. The evaluation of the appearance can be helped by stirring or disturbing the pea protein fermentate (e.g., with a spoon) to observe how the texture and appearance affect the appearance.

[0057] Sensory properties

[0058] The compositions and methods described herein are characterized in that one or more sensory attributes are modulated relative to equivalent pea protein compositions that have not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria / have not been fermented with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria. The modulated sensory attributes can include, but are not limited to, bitterness, sourness, sweetness, pea flavor, green / grassy notes, nutty notes, chalky flavor, acidic notes, and umami flavor.

[0059] For example, relative to equivalent pea protein compositions that have not been contacted with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria / have not been fermented with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria, the pea protein fermentates described herein have increased sweetness, increased sourness, decreased pea flavor, decreased green / grassy notes, increased nutty notes, increased umami flavor, or combinations thereof.

[0060] As used herein, "sensory attribute" refers to the taste, aroma, and / or flavor associated with a given composition that has characteristic properties familiar to a person trained in sensory evaluation. For example, saltiness is associated with sodium chloride, sweetness is associated with sucrose, sourness is associated with citric acid, bitterness is associated with caffeine, and umami is associated with monosodium glutamate (MSG).

[0061] As used herein, "taste" refers to the sensory perception on the tongue. For example, the five basic tastes are sweet, sour, salty, bitter, and umami.

[0062] As used herein, "aroma" refers to the orthonasal olfaction in the nasal cavity.

[0063] As used herein, "flavor" refers to the taste and retronasal olfaction in the nasal cavity.

[0064] As used herein, "off-flavor" refers to a taste or flavor attribute characteristic that is not a characteristic of or not normally associated with the substance or composition described herein, and / or a characteristic taste or flavor associated with an undesirable substance or composition. For example, an off-flavor can be an undesirable taste (such as bitterness), an undesirable mouthfeel (such as astringency, dry mouth), an undesirable flavor (such as rancidity, cardboardy flavor, aftertaste), an inconsistent flavor (e.g., a flavor with uneven onset or intensity, a flavor that may be perceived prematurely or too late), etc.

[0065] As used herein, "plant protein flavor" refers to the characteristic flavor associated with and expected from a plant-based protein when the plant-based protein is used as an ingredient in food and beverage products. For example, plant protein flavors include the beany, pea, corn, hay, green, barnyard, fermented, waxy flavors and combinations thereof that are typically found and expected from plant-based proteins. In general, certain characteristic plant protein flavors can be attributed to certain plant-based proteins. For example, pea protein can be associated with green, pea and hay flavors; soy protein can be associated with beany and hay flavors, corn protein can be associated with corn and hay flavors, and potato protein can be associated with barnyard and fermented flavors.

[0066] A sensory panel can be used to determine, for example, the magnitude of reduction in bitterness or the change in its temporal characteristics. A sensory panel is an essential scientific and reproducible method in the food and beverage industry. A sensory panel involves a group of two or more individual panelists. Panelists are guided by industry-accepted practices to avoid the influence of personal subjectivity and enhance reproducibility. For example, panelists can objectively evaluate the sensory attributes of the product being tested but not provide subjective attributes such as personal preference. In all aspects, a sensory panel can be conducted by two, three, four, five, six or more panelists, where the panelists identify and agree on a sensory attribute lexicon for a given set of samples. After evaluating a particular sample, panelists can assign a numerical intensity score to each attribute using an intensity scale. For example, the intensity scale can range from 0 to 6 (i.e., 0 = not detected, 1 = trace, 2 = slight, 3 = moderate, 4 = distinct, 5 = strong, 6 = extreme), 0 to 9 (i.e., 0 = not detected, 1 = trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = distinct, 7 = strong, 8 = very strong, 9 = extreme), or 0 to 15, where 0 corresponds to the absence of the attribute and 6, 9 or 15 corresponds to the upper extreme occurrence of the attribute, respectively. The panel can use a roundtable consensus approach, or panelists can individually score and evaluate the sensory attributes. Any format may also involve a panel leader who guides the discussion on terminology and guides panelists in evaluating specific products and attributes. In other aspects, a trained sensory panel can be used to evaluate specific attributes using descriptive analysis or temporal intensity methods.

[0067] As used herein, "panelist" refers to highly trained expert tasters, such as those commonly used in sensory methodologies such as descriptive analysis, and / or experienced tasters familiar with the sensory attributes being tested. In some aspects, the panelist can be a trained panelist. A trained panelist has undergone training to understand the terms and sensory phenomena associated with the sensory attributes associated with the test product, and to align on the use of common descriptors (i.e., a sensory lexicon) for those sensory attributes of interest. For example, a trained panelist testing a given composition will understand the terms and sensory attributes associated with that composition, such as saltiness, sourness, bitterness, astringency, mouthfeel, acidity, etc. The trained panelist will be trained on reference samples corresponding to the sensory attributes being tested, and will thus have been calibrated to recognize and quantitatively evaluate such standards. In some aspects, the panelist can be an experienced taster.

[0068] As used herein, "round-robin method" refers to a sensory panel determination methodology in which panelists discuss sensory attributes and intensities and then reach agreement on the intensity scores and attribute characterizations for the specific sensory attributes being determined. A sensory panel using the round-robin method can include two, three, four, five, six or more panelists. The range of the agreement intensity scale can be from 0 to 6 (i.e., 0 = not detected, 1 = trace, 2 = slight, 3 = moderate, 4 = distinct, 5 = strong, 6 = extreme) or 0 to 9 (i.e., 0 = not detected, 1 = trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = distinct, 7 = strong, 8 = very strong, 9 = extreme). For a given set of samples, the panelists will identify and agree on a lexicon of sensory attributes, including (if applicable) reference or standardized samples (also called sensory anchors) for specific sensory attributes. The reference sample for a given sensory attribute will depend on the samples being determined and the sensory attribute lexicon determined by the panel. Those skilled in the art will recognize the appropriate lexicon and reference or standard samples necessary for the sensory evaluation of a given sample.

[0069] In some aspects, after the panelists have agreed on a lexicon for sensory attributes and intensity ratings or have been instructed in the lexicon for sensory attributes and intensity ratings, the panelists independently score and evaluate the samples, including, if applicable, a determination-specific calibration against a reference sample (also referred to as a sensory anchor) for a particular sensory attribute. Examples of common reference samples are described below. The panelists may repeat the evaluation of the samples or may be unaware of the samples they are testing. The samples being tested may be provided to the panelists randomly or in sequential order. In some aspects, the samples may be tested by the panelists using a random balanced order. Then, standard statistical analysis methods are used to evaluate the scores from each panelist to determine the average sensory intensity rating. Those skilled in the art will recognize the appropriate lexicon and reference or standard samples and appropriate statistical analysis methods necessary for the sensory evaluation of a given sample.

[0070] As used herein, "random balanced order" refers to the order in which samples are presented, where the order is random and all possible orders of presenting the samples will be presented among all panelists to eliminate bias for samples tested in a particular order. For example, for a random balanced order of two samples, a given panelist has an equal likelihood of receiving sample 1 before sample 2 and receiving sample 2 before sample 1. In an example with three samples (i.e., sample 1, sample 2, and sample 3), the random balanced order will include an equal likelihood that the panelist receives the samples in the following orders: (i) 1, 2, 3; (ii) 1, 3, 2; (iii) 2, 1, 3; (iv) 2, 3, 1; (v) 3, 2, 1; (vi) 3, 1, 2.

[0071] The sensory attributes of a given composition can be evaluated by comparison with one or more reference or anchor samples. For example, an experienced panelist can use a sodium chloride solution as a saltiness anchor to evaluate the relative intensity of saltiness of a given composition; an experienced panelist can use a sucrose solution as a sweetness anchor to evaluate the relative sweetness intensity of a given composition; an experienced panelist can use a citric acid solution as an acidity anchor to evaluate the relative intensity of acidity of a given composition; an experienced panelist can use a caffeine solution as a bitterness anchor to evaluate the relative bitterness intensity of a given composition; an experienced panelist can use a monosodium glutamate (MSG) solution as an umami anchor to evaluate the relative umami intensity of a given composition. Solutions for evaluating sensory attributes can be provided to experienced panelists, such as 10 mL - 20 mL samples. The experienced panelist dispenses approximately 3 mL - 4 mL of each solution into their own mouth, disperses the solution by moving their tongue, and records the value of the specific sensory attribute being tested. If multiple solutions are to be tested at one time, the panelist can purify their taste buds with water between samples. For example, a round table evaluation of saltiness, sweetness, acidity, umami, etc. can be assigned a scale of 0 to 9, such that a score of 0 indicates no saltiness and a score of 9 indicates extreme saltiness (0 = not detected, 1 = trace, 2 = weak, 3 = slight, 4 = moderate, 5 = medium, 6 = distinct, 7 = strong, 8 = very strong, 9 = extreme). Equivalent scales and methodologies can be used for sweetness, bitterness, acidity, and umami sensory attributes.

[0072] As another example, the saltiness of a composition can be tested by a panel consisting of at least two panelists. The panelists can use a standard range of aqueous sodium chloride solutions corresponding to 0.18 (wt)%, 0.2 (wt)%, 0.35 (wt)%, 0.5 (wt)%, 0.567 (wt)%, 0.6 (wt)%, 0.65 (wt)%, and 0.7 (wt)% having saltiness intensity values corresponding to 2, 2.5, 5, 8.5, 10, 11, 13, and 15, respectively. One of ordinary skill in the art will recognize that the number and range of the standard solutions can vary depending on the sample / composition being tested (e.g., only using solutions corresponding to saltiness intensity values of 2, 2.5, and 5). For each test composition, the panelists will dispense approximately 2 mL - 5 mL (for liquid compositions or solutions prepared with water) or 5 g - 10 g (for solid compositions) of each composition into their own mouths, disperse the composition by moving their tongues / chewing, and record a saltiness intensity value between 0 and 15 for each composition based on comparison with the aforementioned standard sodium chloride solutions. Between tasting compositions, the panelists are able to purify their palates with water. The panelists can also optionally taste the standard 0.18%, 0.2%, 0.35%, 0.5%, 0.567%, 0.6%, 0.65%, and 0.7% sodium chloride solutions between tasting the test solutions to ensure that the recorded saltiness intensity values are accurate relative to the scale of the standard sodium chloride solutions. The temperature at which the test is conducted can be specific to the sample being tested at the start of the test, e.g., the sample can be tested at 22°C (e.g., room temperature), 0°C (e.g., for frozen samples), or between 60°C and 80°C (e.g., for cooked samples of hot food). One of ordinary skill in the art will recognize the appropriate temperature for testing a given sample. This test is referred to herein as the "standardized saltiness intensity test".

[0073] The sourness of the composition can be tested by a panel consisting of at least two panelists. The panelists can use a standard range of aqueous citric acid solutions corresponding to sourness intensity values of 0.035 (wt)%, 0.05 (wt)%, 0.07 (wt)%, 0.15 (wt)%, and 0.2 (wt)% respectively, which correspond to sourness intensity values of 2, 3, 5, 10, and 15. Those skilled in the art will recognize that the number and range of the standard solutions can vary depending on the sample / composition being tested (e.g., only using solutions corresponding to sourness intensity values of 2 and 7). For each test composition, the panelists will dispense approximately 2 mL - 5 mL (for liquid compositions or solutions prepared with water) or 5 g - 10 g (for solid compositions) of each composition into their own mouths, disperse the composition by moving their tongues / chewing, and record a sourness intensity value between 0 and 15 for each composition based on comparison with the aforementioned standard citric acid solutions. Between tasting the compositions, the panelists are able to purify their taste buds with water. The panelists can also optionally taste the standard 0.035%, 0.05%, 0.07%, 0.15%, and 0.2% citric acid solutions between tasting the test solutions to ensure that the recorded sourness intensity values are accurate relative to the scale of the standard citric acid solutions. The temperature at which the test is conducted can be specific to the sample at which the test is initiated, e.g., the sample can be tested at 22°C (e.g., room temperature), 0°C (e.g., for frozen samples), or between 60°C and 80°C (e.g., for cooked samples of hot food). Those skilled in the art will recognize the appropriate temperature for testing a given sample. This test is referred to herein as the "standardized sourness intensity test".

[0074] The bitterness of the composition can be tested by a panel consisting of at least two panelists. The panelists can use a standard range of caffeine solutions corresponding to bitterness intensity values of 2, 3, 4, 5, 10, and 15, namely 0.0125 (wt)%, 0.01875 (wt)%, 0.025 (wt)%, 0.031 (wt)%, 0.07 (wt)%, and 0.12 (wt)%, respectively. Those skilled in the art will recognize that the number and range of the standard solutions can be changed depending on the sample / composition being tested (e.g., only using solutions corresponding to bitterness intensity values of 2, 3, and 5). For each test composition, the panelists will dispense approximately 2 mL - 5 mL (for liquid compositions or solutions prepared with water) or 5 g - 10 g (for solid compositions) of each composition into their own mouths, disperse the composition by moving their tongues / chewing, and record a bitterness intensity value between 0 and 15 for each composition based on comparison with the aforementioned standard caffeine solutions. Between tasting the compositions, the panelists are able to purify their taste buds with water. The panelists can also optionally taste the standard 0.0125%, 0.01875%, 0.025%, 0.031%, 0.07%, and 0.12% caffeine solutions between tasting the test solutions to ensure that the recorded bitterness intensity values are accurate relative to the scale of the standard caffeine solutions. The temperature at which the test is conducted can be specific to the sample at the start of the test. For example, the sample can be tested at 22°C (e.g., room temperature), 0°C (e.g., for frozen samples), or between 60°C and 80°C (e.g., for cooked samples of hot food). Those skilled in the art will recognize the appropriate temperature for testing a given sample. This test is referred to herein as the "standardized bitterness intensity test."

[0075] The sweetness of the composition can be tested by a panel consisting of at least two panelists. The panelists can use a standard range of sucrose solutions at 2 (wt)%, 5 (wt)%, 8 (wt)%, 10% (wt)%, and 15 (wt)% corresponding to sweetness intensity values of 2, 5, 8, 10, and 15, respectively. Those skilled in the art will recognize that the number and range of the standard solutions can be varied depending on the sample / composition being tested (e.g., using only solutions corresponding to sweetness intensity values of 2, 5, and 8). For each test composition, the panelists will dispense approximately 2 mL - 5 mL (for liquid compositions or solutions prepared with water) or 5 g - 10 g (for solid compositions) of each composition into their own mouths, disperse the composition by moving their tongues / chewing, and record a sweetness intensity value between 0 and 15 for each composition based on comparison with the aforementioned standard sucrose solutions. Between tasting compositions, the panelists are able to purify their taste buds with water. The panelists can also optionally taste the standard 2%, 5%, 8%, 10%, and 15% sucrose solutions between tasting the test solutions to ensure that the recorded sweetness intensity values are accurate relative to the scale of the standard sucrose solutions. The temperature at which the test is conducted can be specific to the sample being tested at the start of the test, e.g., the sample can be tested at 22 °C (e.g., room temperature), 0 °C (e.g., for frozen samples), or between 60 °C and 80 °C (e.g., for cooked samples of hot food). Those skilled in the art will recognize the appropriate temperature for testing a given sample. This test is referred to herein as the "standardized sweetness intensity test".

[0076] The umami of the composition can be tested by a panel consisting of at least two panelists. The panelists can use a standard range of 0.75% (by weight) and 0.125% (by weight) monosodium glutamate (MSG) solutions corresponding to umami intensity values of 4 and 6.5, respectively. Those skilled in the art will recognize that the number and range of the standard solutions can be changed according to the sample / composition being tested (e.g., if the expected umami intensity is significantly outside the umami intensity values of 4 - 6.5, additional umami solutions are added). For each test composition, the panelists will dispense approximately 2 mL - 5 mL (for liquid compositions or solutions prepared with water) or 5 g - 10 g (for solid compositions) of each composition into their own mouths, disperse the composition by moving their tongues / chewing, and record an umami intensity value between 0 and 15 for each composition based on comparison with the aforementioned standard MSG solutions. Between tasting the compositions, the panelists are able to purify their palates with water. The panelists can also optionally taste the standard 0.075% and 0.125% MSG solutions between tasting the test solutions to ensure that the recorded umami intensity values are accurate relative to the scale of the standard MSG solutions. The temperature at which the test is conducted can be specific to the sample being tested at the start of the test. For example, the sample can be tested at 22°C (e.g., room temperature), 0°C (e.g., for frozen samples), or between 60°C and 80°C (e.g., for cooked samples of hot food). Those skilled in the art will recognize the appropriate temperature for testing a given sample. This test is referred to herein as the "standardized umami intensity test".

[0077] Control samples are typically used as a reference point or for comparison purposes. The control sample can be a composition such as those described herein, but which has not been fermented or contacted with Leuconostoc citreum bacteria. Similarly, the control sample can be a reference sample having a similar protein composition, sweetness, etc., but made with different ingredients (such as the pea protein ferment described herein). Except for the pea protein ferment, the control sample is otherwise identical and should contain the same components and other ingredients at the same relevant concentrations. Other standard samples are commonly used by sensory panels, such as standard samples for evaluating the intensity of sensory attributes as outlined above.

[0078] This disclosure is not limited to sensory testing by experienced or trained panelists. For example, untrained and inexperienced panelists can be utilized. However, in the case of untrained and inexperienced panelists, a larger number of panelists are required to provide reproducible results, which will typically focus on subjective attributes such as preference or overall liking. Similarly, untrained and inexperienced panelists can be asked to evaluate the relative change in a given sensory attribute between two samples. For example, if a particular sample is more or less salty, more or less sweet, more or less bitter, etc., than a reference sample.

[0079] Exemplary sensory assays and test criteria for additional sensory attributes are described in the embodiments provided in this disclosure.

[0080] Example

[0081] The present invention is further described in detail by reference to the following experimental embodiments. These embodiments are provided for illustrative purposes only and are not intended to be limiting, unless otherwise stated. Accordingly, the present invention should in no way be construed as limited to the following embodiments, but should be construed to cover any and all variations that become apparent from the teachings provided herein.

[0082] Example 1 – Pea protein fermentate with Leuconostoc citreum or Leuconostoc pseudomesenteroides

[0083] Three Leuconostoc citreum strains isolated from different ecosystems were used to ferment pea protein isolate. Each of the three strains B3K7, C22B11, and C18X24 was identified by MALDI-TOF_MS fingerprinting and assigned to the species Leuconostoc citreum (B3K7 and C22B11) or Leuconostoc pseudomesenteroides (C18X24). The MALDI spectra obtained were identical, and the spectrum from B3K7 was used as the representative of the cluster. The B3K7 strain was subjected to 16s rRNA gene sequencing and whole genome sequencing to confirm the species assignment.

[0084] Table 1 outlines the combinations of pea protein cultures and Leuconostoc citreum or Leuconostoc pseudomesenteroides strains used for pea protein fermentation. Each fermentation was inoculated with 3.8 wt% of the indicated Leuconostoc strain. Fermentation was carried out in water. The cultures outlined in Table 1 were fermented at 25 °C for 24 hours without agitation. The syneresis, appearance, taste, and texture of the resulting samples were then evaluated. Syneresis was evaluated by centrifuging the fermented samples at 5,000 g for 10 minutes. The comparison of the syneresis observed in the control starch and protein suspensions with the absence of syneresis observed in Sample 1.2 is shown in Figure 8 in. Appearance was evaluated visually.

[0085] Assays were conducted to characterize the sensory attributes of the samples. The sensory attributes (taste and texture) were evaluated by a panel of 12 individuals experienced in plant protein sensory testing. The experienced panelists evaluated sensory attributes such as, but not limited to, green pea flavor, sourness, chalkiness, nutty flavor, sweetness, and texture. To test each sample, the experienced panelists placed a portion of each sample in their own mouths, dispersed the sample around their mouths, and recorded their observations. Visual evaluations were also made of the texture and appearance observations.

[0086] Photographs of the samples outlined in Table 1 before and after manual stirring for about 2 seconds to 5 seconds are provided in Figure 1 , Figure 2 and Figure 3 .

[0087] Overall, although there is some strain-to-strain variability in the fermented samples, all strains showed a significant viscosity effect, acidification of the samples, and flavor modification that reduced the grassy and green pea flavors of the samples. The fermentate produced using Leuconostoc citreum B3K7 is a starter dough with a very smooth, shiny, and glistening texture, rich in acidic and umami notes. Leuconostoc citreum C22B11 started to thicken faster (about 4 hours to 5 hours), produced less sourness but more nutty notes, and the texture seemed to be thicker, with very firm peaks, and required manual stirring. The sample fermented by Leuconostoc pseudomesenteroides C18X24 has a very viscous and slippery texture, similar to melted cheese (see Figure 4 ).

[0088]

[0089]

[0090] Example 2 - Viscosity and acidification

[0091] Together with a blank pea protein sample containing pea protein isolate, sucrose, and water but not inoculated with any Leuconostoc citreum or Leuconostoc pseudomesenteroides strains, the viscosities of the samples outlined in Table 1 were tested. Viscosity (cP) was measured using a Rapid Visco Analyzer (RVA) at 25 °C and 250 rpm. As shown in Figures 5 to 7 and Table 2, all samples showed a significant increase in viscosity relative to the unfermented pea protein blank. However, there was some strain variability in the absolute increase in viscosity compared to the blank.

[0092] The samples were also subjected to two consecutive freeze-thaw cycles, and the viscosity was measured after freeze-thawing, and it was found to be consistent with the samples before freezing. Visual analysis also showed that there was no change or loss in thickness after freezing and thawing.

[0093] The pH level of each fermentate varied based on the strain, protein concentration, and sucrose concentration. Overall, the pH increased with increasing protein concentration (decreased acidification). This effect was most obvious for the B3K7 strain. For strains C22B11 and C18X24, although the pH increased with increasing protein concentration, the pH of the samples produced with more than 6% protein was less significant.

[0094] Table 2 .

[0095]

[0096]

[0097] Example 3 – Composition analysis

[0098] Samples 1.2 were selected for further compositional analysis based on appearance, scoopability, and taste. Sample 1.2 was analyzed by UPLC-RI to quantify simple carbohydrates and by HPLC-RI to quantify sugar alcohols and organic acids. The sucrose (15%) present at the start of fermentation was completely utilized by the end of fermentation, and fructose, α-glucan, and mannitol were produced. Lactic acid and acetic acid were also present in the resulting fermentation. Although the α-glucan concentration was not quantified independently, this value was obtained by subtracting the other identified carbohydrates from the total carbohydrates present. The compositional analysis of sample 1.2 is reported in Table 3.

[0099] Table 3 .

[0100] Component % Sample 1.2 Protein 3.3 Carbohydrate 16.1 Glucose 0.0 Fructose 6.0 Sucrose 0.0 α-Glucan 6.5 Polyol 2.0 Dietary fiber 0.2 Organic acid 1.4 Fat 0.3 Ash 0.2 Water 79.1 Total solids 19

[0101] Example 4 – Pea protein dessert

[0102] The appearance and consistency of the pea protein fermentate compositions of samples 1.2, 1.10, and 1.18 were compared to the reference vegan dessert formulations outlined in Table 4. The compositional analysis of the reference vegan dessert formulations is provided in Table 5. For comparison, the reference vegan dessert was formulated to match the protein content and sweetness of the pea protein fermentate of sample 1.2. The reference vegan dessert formulation was prepared by mixing the dry ingredients (pea protein, sucrose, starch, and carrageenan) and adding the mixed dry ingredients to water at room temperature (about 25°C) while stirring at 800 rpm. After hydrating the mixed dry ingredients, the entire mixture was pasteurized by heating to 95°C. The pasteurized mixture was cooled to between 50°C and 70°C and then refrigerated (4°C) overnight. The sensory attributes and aspects of the samples were analyzed the next day.

[0103] A visual comparison of the appearance of the reference vegan dessert with the vegan desserts prepared with samples 1.2, 1.10, and 1.18 is provided in Figure 9 and Figure 10 . Overall, compared to the reference vegan dessert samples, the vegan desserts prepared with samples 1.2 and 1.10 had a smoother texture and a more glossy and shiny appearance.

[0104] In addition, the sensory attributes of a reference vegan dessert and Sample 1.2 were compared. Relative to the reference vegan dessert, when sampled by a panel of 4 individuals experienced in sensory evaluation, Sample 1.2 was found to be spicier, less bitter, less metallic, less beany, less astringent, brighter, and have a better mouthfeel. See Figure 11 .

[0105] Table 4 .

[0106] Ingredient Weight % Pea protein isolate 4.0 Sucrose 7.0 Hydroxypropylated starch 5.0 Carrageenan 0.015 Water 84.0

[0107] Table 5 .

[0108] Component % Reference vegan dessert Protein 3.3 Carbohydrate 12.5 Sucrose 7.0 Dietary fiber 0.2 Fat 0.3 Ash 0.2 Water 84 Total solids 16

[0109] Example 5 – Comparative pea protein fermentation

[0110] Fermentation of pea protein by three Leuconostoc citreum strains and one Leuconostoc pseudomesenteroides strain was compared to fermentation by a dairy-derived lactic acid bacteria culture (trade name “YO-MIX 433”) containing Streptococcus thermophilus and Lactobacillus bulgaricus. This bacterial culture is known in the art as a yogurt culture and is sold by . Fermentation of pea protein using YO-MIX 433 was previously described in US2020 / 296982.

[0111] In this example, pea protein fermentation was carried out using four different strains / cultures and three different fermentation protocols, as outlined in Tables 6 and 7.

[0112] The appearance, texture, and syneresis of the resulting samples from the cultures outlined in Table 7 were evaluated. Appearance was evaluated visually. The results are summarized in Tables 8 and 9 and Figures 12 to 16 .

[0113] Overall, compared to the YO-MIX 433 culture, the C18X1, B3K7, C22B11, and C18X24 strains produce pea protein fermentates with higher viscosities. Under the experimental conditions of Example 1 and the experimental conditions of US2020 / 296982, the C18X1, B3K7, C22B11, and C18X24 strains produce products with higher viscosities and lower residual sucrose. When the fermentation conditions of US2020 / 296982 are used together with the 25°C fermentation temperature of Example 1, considering the lower temperature preference of the current strains, the C18X1, B3K7, C22B11, and C18X24 strains again show products with higher viscosities compared to the YO-MIX 433 culture. Additionally, the products fermented with the C18X1, B3K7, C22B11, and C18X24 strains have significantly different appearances from the products of the YO-MIX 433 culture. For example, samples 5.1 - 5.4 are smooth, delicate, thick, or elastic and stretchable, but sample 5.5 shows high syneresis and becomes thin when stirred. Similarly, samples 5.11 - 5.14 are low-syneresis gels that become smoother and more delicate with stirring, while sample 5.15 has high syneresis and becomes thin when stirred. Although the bacteria in the YO-MIX433 culture are characterized as lactic acid bacteria, they do not produce pea protein fermentates with a smooth, delicate texture, low syneresis, and high viscosity like the Leuconostoc citreum and Leuconostoc pseudomesenteroides bacteria described herein.

[0114] Table 6 .

[0115]

[0116]

[0117] Table 7 .

[0118]

[0119] Table 8 .

[0120]

[0121] Table 9 .

[0122]

[0123]

[0124] Example 6 – Comparative plant protein fermentation

[0125] In this example, nine different bacterial strains were used to ferment three different plant-based proteins: pea protein, vital wheat gluten, and corn protein. Five additional publicly available strains that were tested, in addition to the four strains described herein, are outlined in Table 10. Each fermentation condition was inoculated with 3.8 wt% of the indicated bacterial strain. Fermentation was carried out in water. The cultures outlined in Table 9 were fermented at 25 °C for 24 hours without agitation. Then the viscosity of the resulting samples was evaluated. The results are shown in Table 11 and Figures 17 to 21 in.

[0126] Table 10 .

[0127] Species Strain Deposit collection code Leuconostoc citreum TCV-482 NRRL B742 / ATTC 13146 Leuconostoc mesenteroides TCV-447 NRRL B21297 Leuconostoc mesenteroides TCV-474 NRRL B30821 Leuconostoc mesenteroides TCV-464 NRRL B512-F Leuconostoc mesenteroides TCV-487 NRRL B1299 / ATTC 11449

[0128] Table 11 .

[0129]

[0130]

[0131] Terms describing the present invention

[0132] Clause 1. A composition comprising:

[0133] pea protein

[0134] Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria; and

[0135] sucrose.

[0136] Clause 2. A method for fermenting pea protein, the method comprising:

[0137] (i) fermenting the composition according to Clause 1 for a period of time under conditions sufficient to produce a fermented pea protein product.

[0138] Clause 3. A method for fermenting pea protein, the method comprising:

[0139] (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for a period of time under conditions sufficient to produce a fermented pea protein product.

[0140] Clause 4. The composition or method according to any one of Clauses 1 to 3, wherein the composition comprises between 1 wt% and 20 wt%, between 2 wt% and 18 wt%, or between 4 wt% and 15 wt% of pea protein, and / or between 1 wt% and 30 wt%, between 2 wt% and 25 wt%, or between 5 wt% and 20 wt% of sucrose.

[0141] Clause 5. The composition or method according to any one of Clauses 1 to 4, wherein the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria are selected from the group consisting of: Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number LMG P-32799), and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195).

[0142] Clause 6. The method according to any one of Clauses 2 to 5, wherein the composition is fermented for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; and / or the composition is fermented at a temperature between 20 °C and 30 °C, between 22 °C and 28 °C, between 24 °C and 26 °C, or at about 25 °C.

[0143] Clause 7. The method according to any one of Clauses 2 to 6, the method further comprising the step of stirring the fermented pea protein product during or after step (i).

[0144] Clause 8. The method according to any one of Clauses 2 to 7, wherein

[0145] the method is a method for increasing the viscosity of pea protein, and the fermented pea protein product has a higher viscosity compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc bacteria; and / or

[0146] the method is a method for changing the appearance of pea protein, and the fermented pea protein product has a more shiny, more lustrous, more viscous, and / or more gel-like appearance compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc bacteria; and / or

[0147] or

[0148] the method is a method for changing one or more sensory attributes of a pea protein composition, and the fermented pea protein product has an increased sour taste, a reduced green pea flavor,

[0149] an increased sweet taste, and / or a reduced grassy flavor compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc bacteria.

[0150] Clause 9. A composition comprising a fermented pea protein product obtained by the method according to any one of Clauses 2 to 8.

[0151] Clause 10. The composition according to Clause 9, wherein the composition comprises fructose, α-glucan, polyol and / or organic acid, and optionally, wherein the composition does not contain sucrose and / or wherein the composition does not contain starch.

[0152] Clause 11. The composition according to Clause 10, wherein the composition comprises α-glucan in an amount between 1% and 20% by weight, between 2% and 15% by weight, or between 5% and 10% by weight.

[0153] Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for increasing the viscosity of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose.

[0154] Clause 13. A method for obtaining a shiny pea protein product, the method comprising:

[0155] (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria at a temperature between 20°C and 30°C, between 22°C and 28°C, between 24°C and 26°C, or at about 25°C for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours;

[0156] (ii) stirring the contacted pea protein composition during or after step (i); and

[0157] (iii) obtaining the stirred fermented pea protein product, which has a shinier appearance than the pea protein composition before contact with the Leuconostoc bacteria.

[0158] Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for increasing the glossiness of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose.

[0159] Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for altering one or more sensory properties of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose, wherein the sensory properties are selected from the group consisting of sour taste, green pea flavor, sweet taste, and grassy flavor.

[0160] Clause 16. The use or method according to any one of Clauses 12 to 15, wherein the pea protein concentration is between 1% and 20% by weight, between 2% and 18% by weight, or between 4% and 15% by weight, and / or the sucrose concentration is between 1% and 30% by weight, between 2% and 25% by weight, or between 5% and 20% by weight.

[0161] Clause 17. A composition comprising:

[0162] Fermented pea protein; and

[0163] Inactive Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria;

[0164] wherein the composition does not contain added sucrose and / or wherein the composition does not contain added starch.

[0165] Clause 18. Isolated Leuconostoc citreum or Leuconostoc pseudomesenteroides bacterial cells selected from the group consisting of Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number P-32799), and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195).

Claims

1. A composition, the composition comprising: pea protein Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria; and sucrose.

2. A method for fermenting pea protein, the method comprising: (i) fermenting the composition according to claim 1 for a period of time under conditions sufficient to produce a fermented pea protein product.

3. The composition or method according to any one of claims 1 to 2, wherein the composition comprises between 1% and 20% by weight, between 2% and 18% by weight, or between 4% and 15% by weight of pea protein, and / or between 1% and 30% by weight, between 2% and 25% by weight, or between 5% and 20% by weight of sucrose.

4. The composition or method according to any one of claims 1 to 3, wherein the Leuconostoc citreum bacteria are selected from the group consisting of: Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BMCC accession number LMG P-32799) and Leuconostoc pseudomesenteroides C18X24 (BCCM accession number LMG P-33195).

5. The method according to any one of claims 2 to 4, wherein the composition is fermented for at least 6 hours, at least 12 hours, at least 18 hours or at least 24 hours; and / or the composition is fermented at a temperature between 20°C and 30°C, between 22°C and 28°C, between 24°C and 26°C or at about 25°C.

6. The method according to any one of claims 2 to 5, the method further comprising the step of stirring the fermented pea protein product during or after step (i).

7. The method according to any one of claims 2 to 6, wherein the method is a method for increasing the viscosity of pea protein, and the fermented pea protein product has a higher viscosity compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc citreum bacteria; and / or the method is a method for changing the appearance of pea protein, and the fermented pea protein product has a more shiny, more viscous and / or more gel-like appearance compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria; and / or the method is a method for changing one or more sensory properties of a pea protein composition, and the fermented pea protein product has an increased sour taste, a reduced green pea flavor, an increased sweet taste and / or a reduced grassy flavor compared to an equivalent pea protein composition that has not been contacted with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria.

8. A composition preferably obtained by the method according to any one of claims 2 to 7, the composition comprising fermented pea protein and Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria.

9. The composition according to claim 8, wherein the composition comprises fructose, α-glucan, polyol and / or organic acid, and optionally, - wherein the composition does not contain sucrose; and / or - wherein the composition does not contain added starch.

10. The composition according to claim 9, wherein the composition comprises α-glucan between 1% and 20% by weight, between 2% and 15% by weight, or between 5% and 10% by weight.

11. The composition according to claim 8, 9 or 10, wherein the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria are non-viable.

12. Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for increasing the viscosity of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose.

13. Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for increasing the glossiness of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose.

14. Use of Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria for altering one or more sensory properties of pea protein by fermenting the pea protein with the Leuconostoc citreum or Leuconostoc pseudomesenteroides bacteria in the presence of sucrose, wherein the sensory properties are selected from the group consisting of sour taste, green pea flavor, sweet taste and grass flavor.

15. The use according to any one of claims 12 to 14, wherein the pea protein concentration is between 1% and 20% by weight, between 2% and 18% by weight, or between 4% and 15% by weight, and / or the sucrose concentration is between 1% and 30% by weight, between 2% and 25% by weight, or between 5% and 20% by weight.

16. Isolated Leuconostoc citreum or Leuconostoc pseudomesenteroides bacterial cells, which are selected from the group consisting of Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), Leuconostoc citreum C18X1 (BCCM accession number LMG P-32799) and Leuconostoc citreum C18X24 (BCCM accession number LMG P-33195).

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