Leuconostoc citreum, use thereof for producing fermented pure vegetarian dessert, in particular comprising fermented pea protein, and pure vegetarian dessert comprising fermented pea protein and said strain

Abstract

Described herein are compositions and methods for producing pure vegetarian dessert compositions comprising a pea protein ferment. A starting composition comprising pea protein, leuconostoc citreum bacteria and sucrose can be fermented for a period of time under conditions suitable for producing a pea protein ferment. The resulting pea protein leavening will have increased viscosity, improved appearance and improved sensory properties relative to equivalent pea protein compositions that have not been contacted with or fermented by the leuconostoc citrinum bacteria. The pea protein ferment is incorporated into a pure vegetarian dessert composition.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of European Priority Application No. 22206229.1, filed on November 8, 2022, which is incorporated herein by reference in its entirety. Background of the invention

[0003] With the increasing public interest in plant - based proteins, pea protein has found an increasingly wide range of applications in the food and beverage industry. 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 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 that can alter 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 properties. Summary of the invention

[0006] The present disclosure provides a vegan dessert composition comprising a fermented pea protein composition; wherein the fermented pea protein composition is made by contacting pea protein with Leuconostoc citreum B3K7 or Leuconostoc citreum C22B11 in the presence of sucrose; and optionally, a sweetener, a flavorant, or a combination thereof. The vegan dessert composition can be free of added starch. The vegan dessert composition can comprise a sweetener, which includes steviol glycosides, mogrosides, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

[0007] The present disclosure also provides a method for producing a vegan dessert composition, the method comprising: (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum B3K7 or Leuconostoc citreum C22B11 for a period of time under conditions sufficient to produce a fermented pea protein product; (ii) optionally, inactivating the Leuconostoc citreum bacteria in the fermented pea protein (e.g., by pasteurization, heat killing, irradiation, or chemical treatment); and (iii) adding a sweetener, a flavorant, or a combination thereof to the fermented pea protein product, thereby producing a vegan pudding composition. 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% 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. The pea protein 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 pea protein 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 additionally comprise the step of stirring the contacted pea protein composition during or after step (i). The sweetener may comprise steviol glycosides, mogrosides, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

[0008] The present disclosure also provides an isolated Leuconostoc citreum B3K7 bacterial cell and an isolated Leuconostoc citreum C22B11 bacterial cell.

[0009] The present disclosure also provides the use of Leuconostoc citreum B3K7 bacterial cells and / or Leuconostoc citreum C22B11 bacterial cells for producing a vegan dessert composition.

[0010] The present disclosure also provides a composition comprising fermented pea protein, inactivated Leuconostoc citreum B3K7 and / or Leuconostoc citreum C22B11 bacteria; wherein the composition is free of added sucrose and / or wherein the composition is free of added starch. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] This patent or application contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Patent Office upon request and payment of the necessary fee.

[0012] The drawings generally illustrate, by way of example, and not by way of limitation, the various aspects described herein.

[0013] Figure 1Pictures showing the visual appearance of fermentation samples 1.1 - 1.8 before and after agitation, as outlined in Example 1.

[0014] Figure 2 Pictures showing the visual appearance of fermentation samples 1.9 - 1.16 before and after agitation, as outlined in Example 1.

[0015] Figure 3 Pictures showing the visual appearance of fermentation samples 1.17 - 1.24 before and after agitation, as outlined in Example 1.

[0016] Figure 4 Still frames from a video showing the appearance of comparative samples 1.17 and 1.24.

[0017] Figure 5 Graph showing the viscosity curve of Leuconostoc citreum B3K7 fermentates of samples 1.1 - 1.8.

[0018] Figure 6 Graph showing the viscosity curve of Leuconostoc citreum C22B11 fermentates of samples 1.9 - 1.16.

[0019] Figure 7 Graph showing the viscosity curve of Leuconostoc citreum C18X24 fermentates of samples 1.17 - 1.24.

[0020] Figure 8 Graph showing the reduction in syneresis for sample 1.2 relative to a control starch and protein suspension sample.

[0021] Figure 9 Graph showing a comparison of the appearance of a reference vegan dessert formulation made with pea protein, starch, and carrageenan with the appearance of samples 1.2 and 1.10.

[0022] Figure 10 Graph showing the appearance and consistency of a reference vegan dessert formulation made with pea protein, starch, and carrageenan compared to samples 1.2, 1.10, and 1.18.

[0023] Figure 11 Graph showing the relative change in sensory attributes when sample 1.2 is compared to the reference vegan dessert formulation outlined in Example 4.

[0024] Figure 12 Graph showing the viscosity of the samples outlined in Example 5.

[0025] Figure 13 Graph showing the viscosity curve of the Sample A of Scheme A from Example 5.

[0026] Figure 14Shows the viscosity curve of the sample of Scheme B from Example 5.

[0027] Figure 15 Shows the viscosity curve of the sample of Scheme C from Example 5.

[0028] Figure 16 Shows the residual sucrose concentration of the samples outlined in Example 5.

[0029] Figure 17 Shows the viscosity of the samples outlined in Example 6.

[0030] Figure 18 Shows the viscosity curve of the 4 wt% pea protein fermentation sample from Example 6.

[0031] Figure 19 Shows the viscosity curve of the 15 wt% pea protein fermentation sample from Example 6.

[0032] Figure 20 Shows the viscosity curve of the vital wheat gluten fermentation sample from Example 6.

[0033] Figure 21 Shows the viscosity curve of the zein fermentation sample from Example 6. Detailed Description

[0034] Certain aspects of the subject matter disclosed by the present invention will now be specifically referred to, examples of which are partially shown in the accompanying drawings. Although the disclosed subject matter will be described in connection with the recited claims, it should be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0035] 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 incorporated by reference individually. If there is an inconsistency in the usage between this document and those incorporated by reference in this way, 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.

[0036] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also all individual numerical values or sub-ranges subsumed within that range as if each numerical 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 individual values within the specified range (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%). 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".

[0037] Unless expressly indicated otherwise, ppm (parts per million), percentages, and ratios are based on weight. Weight-based percentages are also referred to hereinafter as weight % or (wt) %.

[0038] 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 alteration of one or more sensory attributes relative to the protein prior to fermentation. Generally, pea protein is fermented with Leuconostoc citreum bacteria.

[0039] Fermentate

[0040] The present disclosure relates to compositions comprising pea protein, Leuconostoc citreum bacteria, and sucrose, and methods of using such compositions to produce fermented pea protein products.

[0041] Generally, the starting composition will include pea protein (i.e., protein extracted and / or derived from the seeds or pod fruits of Pisum sativum). The pea protein can be from any suitable source. The 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 A suitable pea protein composition 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.

[0042] The starting composition further contains sucrose. The composition can contain between 1 wt% and 30 wt%, between 2 wt% and 25 wt%, or between 5 wt% and 20 wt% sucrose. 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% sucrose. The sucrose can be from any suitable source. Those skilled in the art will recognize suitable sources, including commercially available sources or sucrose.

[0043] The starting composition can contain a weight ratio of pea protein and sucrose 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.

[0044] 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.

[0045] In the priority application EP 22206229.1 filed on November 8, 2022, strain C18X24 was incorrectly represented as strain C18X1 in the examples and throughout the specification, drawings, and claims. The appropriate correction is made herein. Accordingly, data using strain C18X24 are 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 are provided herein based on strain C18X1, and this strain is different from the incorrectly labeled strain in the priority application.

[0046] 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 bacteria for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours 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 in the presence of sucrose.

[0047] In this text, "fermented pea protein product" and "pea protein fermentate" are used interchangeably and refer to a composition produced by the microbial fermentation of pea protein and include (i) the pea protein; (ii) metabolites produced by the microorganism during the fermentation of pea protein; (iii) non-living microorganisms used in the fermentation process; and (iv) water. The pea protein fermentate may include metabolites such as, but not limited to, fructose, α-glucan, polyols, organic acids, and combinations thereof. For example, the pea protein fermentate may include α-glucan in an amount between 1 wt% and 20 wt%, between 2 wt% and 15 wt%, or between 5 wt% and 10 wt%. The pea protein fermentate may include fructose in an amount between 1 wt% and 20 wt%, between 2 wt% and 15 wt%, or between 5 wt% and 10 wt%. The pea protein fermentate may include polyols in an amount between 0.1 wt% and 10 wt%, between 0.5% and 8%, or between 1 wt% and 5 wt%. The pea protein fermentate may include organic acids in an amount between 0.1 wt% and 10 wt%, between 0.5% and 8%, or between 1 wt% and 5 wt%. The pea protein fermentate may include dietary fiber in an amount between 0.01 wt% and 5 wt%, between 0.05 wt% and 2 wt%, or between 0.1 wt% and 1 wt%. In one example, the pea protein fermentate may include fructose between 5 wt% and 10 wt%, α-glucan between 5 wt% and 10 wt%, polyols between 1 wt% and 5 wt%, organic acids between 1 wt% and 5 wt%, dietary fiber between 0.1 wt% and 1 wt%, protein, fat, and water.

[0048] 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.

[0049] The pea protein fermentate can be processed using an inactivation step in which the microorganisms are made 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 may additionally or alternatively undergo a physical method by which the microorganisms are separated, such as by filtration.

[0050] Although sucrose is used to produce 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 may be utilized by Leuconostoc citreum during fermentation such that the resulting pea protein fermentate is free of sucrose. In one example, the pea protein fermentate can include fructose between 5 wt% and 10 wt%, alpha-glucan between 5 wt% and 10 wt%, polyols between 1 wt% and 5 wt%, organic acids between 1 wt% and 5 wt%, dietary fiber between 0.1 wt% and 1 wt%, 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 such that all of the sucrose in the starting composition is depleted and no additional sucrose is added to the resulting pea protein fermentate.

[0051] 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 the microorganism during fermentation, but no other starch component ingredients are added.

[0052] Additionally, the pea protein fermentate can be agitated to form an agitated pea protein fermentate. The pea protein fermentate can be agitated manually or mechanically. The pea protein fermentate can be agitated 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.

[0053] 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 produce a pea protein fermentate with a slightly higher pH value.

[0054] Generally, the pea protein ferment described herein is characterized by an increase in viscosity relative to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum bacteria / has not been fermented with Leuconostoc citreum bacteria. When measured after stirring at 250 rpm and 25 °C for 5 minutes, the viscosity of the pea protein ferment 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 ferment with a higher viscosity. Similarly, a higher concentration of sucrose in the starting composition will result in a pea protein ferment with a higher viscosity. Thus, as is apparent 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 ferment with a particular desired viscosity.

[0055] The pea protein ferment described herein is characterized by a change in visual and physical appearance relative to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum bacteria / has not been fermented with Leuconostoc citreum bacteria. For example, the pea protein ferment can be glossier, shinier, stickier, and / or have a more gel-like appearance compared to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum bacteria / has not been fermented with Leuconostoc citreum bacteria. A "glossy" appearance of the pea protein ferment is an appearance in which the pea protein ferment is smooth (i.e., no lumps or granules are visible in appearance) and shiny. A "shiny" appearance is an appearance in which the surface of the pea protein ferment reflects light. A "sticky" appearance is an appearance in which the pea protein ferment appears to be soft and sticky. A "gel-like" appearance is an appearance in which the pea protein ferment appears to be thick, slightly sticky, and somewhat firm / solid. The appearance of the pea protein ferment can be evaluated with the naked eye, or it can be assisted by mechanically measuring one of the above traits. Evaluation of the appearance can be helped by stirring or disturbing the pea protein ferment (e.g., with a spoon) to observe how the texture and appearance affect the appearance.

[0056] Sensory property

[0057] The compositions and methods described herein are characterized by the modulation of one or more sensory attributes relative to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum bacteria / has not been fermented with Leuconostoc citreum 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.

[0058] For example, relative to an equivalent pea protein composition that has not been contacted with Leuconostoc citreum bacteria / has not been fermented with Leuconostoc citreum bacteria, the pea protein fermentate described herein has an increased sweetness, an increased sourness, a reduced pea flavor, a reduced green / grassy note, an increased nutty note, an increased umami flavor, or a combination thereof.

[0059] 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).

[0060] 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.

[0061] As used herein, "aroma" refers to the retronasal olfaction in the nasal cavity.

[0062] As used herein, "flavor" refers to the taste and retronasal olfaction in the nasal cavity.

[0063] As used herein, "off - flavor" refers to a taste or flavor attribute characteristic that is not a characteristic or not normally associated with the substance or composition as described herein, and / or a characteristic taste or flavor associated with an undesired substance or composition. For example, an off - flavor can be an undesired taste (such as bitterness), an undesired mouthfeel (such as astringency, dry mouth), an undesired flavor (such as rancidity, cardboardy taste, aftertaste), an inconsistent flavor (e.g., a flavor with uneven onset or intensity, a flavor that may be perceived too early or too late), etc.

[0064] 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, barny, fermented, waxy flavors, and combinations thereof that are typically found and expected from plant - based proteins. Generally, 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. Soybean 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 barny and fermented flavors.

[0065] A sensory panel can be used to determine, for example, the magnitude of a reduction in bitterness or a 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 according to 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 preferences. 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 score and evaluate the sensory attributes individually. Any form can also involve a panel leader who guides discussions 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 time-intensity methods.

[0066] 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, panelists can be trained panelists. Trained panelists have 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., sensory lexicon) for those sensory attributes of interest. For example, trained panelists testing a given composition will understand the terms and sensory attributes associated with that composition, such as saltiness, sourness, bitterness, astringency, mouthfeel, acidity, etc. Trained panelists will be trained on reference samples corresponding to the sensory attributes being tested and have thus been calibrated to recognize and quantitatively evaluate such standards. In some aspects, panelists can be experienced tasters.

[0067] 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 ratings and attribute characterizations for the specific sensory attributes being determined. A sensory panel using the round-robin method may 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 from 0 to 9 (i.e., 0 = not detected, 1 = trace, 2 = weak, 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 dictionary of sensory attributes, including, if applicable, reference or standardized samples (also called sensory anchors) for the specific sensory attributes. The reference sample for a given sensory attribute will depend on the samples being determined and the dictionary of sensory attributes determined by the panel. Those skilled in the art will recognize the appropriate dictionary and reference or standard samples necessary for the sensory evaluation of a given sample.

[0068] In some aspects, after the panelists have reached agreement on or been instructed in a dictionary of sensory attributes and intensity ratings, the panelists independently score and evaluate the samples, including, if applicable, determination-specific calibration against a reference sample (also called a sensory anchor) for the specific sensory attribute. Examples of common reference samples are described below. The panelists may repeat the evaluation of the samples or 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. The scores from each panelist are then evaluated using standard statistical analysis methods to determine the average sensory intensity rating. Those skilled in the art will recognize the appropriate dictionary and reference or standard samples and appropriate statistical analysis methods necessary for the sensory evaluation of a given sample.

[0069] 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 will receive 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.

[0070] 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 the experienced panelists, such as 10 mL - 20 mL samples. The experienced panelists dispense approximately 3 mL - 4 mL of each solution into their own mouths, disperse the solution by moving their tongues, and record the values of the specific sensory attributes being tested. If multiple solutions are to be tested at one time, the panelists 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 from 0 to 9, where a score of 0 indicates no saltiness, 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.

[0071] 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)% with saltiness intensity values corresponding to 2, 2.5, 5, 8.5, 10, 11, 13, 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., only using solutions corresponding to the 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 taste buds 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). Those skilled 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".

[0072] 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 0.035 (wt)%, 0.05 (wt)%, 0.07 (wt)%, 0.15 (wt)%, and 0.2 (wt)% of citric acid with sourness intensity values corresponding to 2, 3, 5, 10, and 15, respectively. Those skilled in the art will recognize that depending on the sample / composition being tested, the number and range of the standard solutions can be changed (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".

[0073] The bitterness of the composition can be tested by a panel consisting of at least two expert panelists. The expert panelists can use a standard range of caffeine solutions corresponding to 0.0125 (wt)%, 0.01875 (wt)%, 0.025 (wt)%, 0.031 (wt)%, 0.07 (wt)%, and 0.12 (wt)% of bitterness intensity values of 2, 3, 4, 5, 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., only using solutions corresponding to bitterness intensity values of 2, 3, and 5). For each test composition, the expert 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 a comparison with the aforementioned standard caffeine solutions. Between tasting the compositions, the expert panelists are able to purify their taste buds with water. The expert 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."

[0074] 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 2 (wt)%, 5 (wt)%, 8 (wt)%, 10% (wt)%, and 15 (wt)% sucrose solutions corresponding to sweetness intensity values of 2, 5, 8, 10, and 15, respectively. One 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 a comparison with the aforementioned standard sucrose solutions. Between tasting the 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 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). One 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".

[0075] The umami of the composition can be tested by a panel consisting of at least two panelists. The panelists can use the standard ranges 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 depending on 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 taste buds 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".

[0076] 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). The control sample is otherwise the same except for the pea protein ferment 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.

[0077] The present 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 greater 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.

[0078] Examples of additional sensory attributes and test criteria for sensory determination are described in the examples provided in this disclosure.

[0079] Dessert composition

[0080] The pea protein ferment described herein can be used to prepare dessert compositions. The dessert compositions can be yogurt, pudding, cheesecake, or similar creamy spoonable desserts. The dessert compositions can be vegan dessert compositions. As used herein, "vegan" refers to a composition that does not contain any products or ingredients of animal or animal origin.

[0081] Generally, the dessert compositions include the pea protein ferment described herein. The dessert compositions can additionally contain one or more additional ingredients, including but not limited to sweeteners, flavorings, seasonings, lipid compositions, water, fiber, starch, hydrocolloids, lecithin, preservatives, acids, and combinations thereof.

[0082] The dessert compositions described herein can contain one or more lipid compositions, such as fats, oils, or combinations thereof. Generally, fat refers to a lipid composition that is solid at room temperature, while oil is liquid at room temperature. The lipid composition can contain saturated fatty acids (also referred to as "saturated fats"), unsaturated fatty acids (also referred to as "unsaturated fats"), or combinations thereof. The lipid composition can include but is not limited to vegetable oil, coconut oil, palm oil, sunflower oil, soybean oil, rapeseed oil, or combinations thereof. Depending on the type of dessert, the dessert composition can additionally contain between 1 wt% and 80 wt%, between 1 wt% and 70 wt%, between 1 wt% and 10 wt%, between 1 wt% and 5 wt%, between 5 wt% and 30 wt%, between 10 wt% and 25 wt%, between 10 wt% and 75 wt%, or between 15 wt% and 70 wt% of the lipid composition. One of ordinary skill in the art will understand the appropriate lipid composition inclusion rate for a given dessert composition.

[0083] The dessert compositions can contain water. For example, depending on the type of dessert composition, the dessert composition can additionally contain between 1 wt% and 80 wt%, between 5 wt% and 75 wt%, between 15 wt% and 70 wt%, between 45 wt% and 65 wt%, between 50 wt% and 60 wt%, between 1 wt% and 20 wt%, or between 5 wt% and 15 wt% of water. Those skilled in the art will recognize the appropriate amount of water (if any) included in a given dessert composition.

[0084] The dessert composition may contain fiber. The fiber may include but is not limited to pectin, apple fiber, psyllium, flax fiber, rice bran extract, konjac flour, and the like. The dessert composition may contain fiber in an amount between 0.01 (weight)% and 3 (weight)%, between 0.05 (weight)% and 2 (weight)%, or between 0.1 (weight)% and 2 (weight)%. The dessert composition may contain fiber in an amount of up to 0.5 (weight)%, up to 1 (weight)%, up to 1.5 (weight)%, up to 2 (weight)%, up to 2.5 (weight)%, or up to 3 (weight)%.

[0085] The dessert composition may contain starch. The starch may include pregelatinized starch, modified starch, or a combination thereof. The starch may include but is not limited to corn starch, potato starch, tapioca starch, and the like. The dessert composition may contain starch in an amount between 0.5 (weight)% and 25 (weight)%, between 1.0 (weight)% and 20 (weight)%, or between 2 (weight)% and 18 (weight)%. Alternatively, the dessert composition may be free of starch. Without wishing to be bound by any particular theory, method, or mode of action, it is believed that including the pea protein ferment described herein in the dessert composition will reduce or eliminate the need to include starch while maintaining the same or similar physical properties. Thus, by having the same viscosity and / or other physical properties, the dessert composition may contain at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% less starch compared to an equivalent dessert composition lacking the pea protein ferment.

[0086] The dessert composition may contain hydrocolloids. For example, the dessert composition may contain guar gum, xanthan gum, locust bean gum, carrageenan, cellulose, konjac gum, and combinations thereof. The dessert composition may contain hydrocolloids in an amount between 0.01 weight% and 5 weight%, between 0.05 weight% and 4.5 weight%, between 0.1 weight% and 4.0 weight%, or between 0.5 weight% and 3.8 weight%. The dessert composition may contain hydrocolloids in an amount of up to 5 weight%, up to 4.5 weight%, up to 4.0 weight%, up to 3.8 weight%, up to 3.5 weight%, up to 2.5 weight%, up to 2.0 weight%, or up to 1.0 weight%. Alternatively, the dessert composition may be free of hydrocolloids. Without wishing to be bound by any particular theory, method, or mode of action, it is believed that including the pea protein ferment described herein in the dessert composition will reduce or eliminate the need to include hydrocolloids while maintaining the same or similar physical properties. Thus, by having the same viscosity and / or other physical properties, the dessert composition may contain at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% less hydrocolloids compared to an equivalent dessert composition lacking the pea protein ferment.

[0087] The dessert composition may contain lecithin. The dessert composition may contain lecithin in an amount between 0.01% and 10% by weight, between 0.05% and 8.0% by weight, or between 0.1% and 5% by weight.

[0088] The dessert composition may contain a preservative. For example, the dessert composition may contain a preservative such as, but not limited to, potassium sorbate. The dessert composition may contain a preservative in an amount up to 0.1% by weight, up to 0.5% by weight, or up to 1.0% by weight of the dessert composition.

[0089] The dessert composition may contain a flavoring agent or a condiment. For example, the dessert composition may contain a natural or artificial flavoring agent and / or a condiment. The condiment and / or flavoring agent may include, but not be limited to, a sweetening agent, salts (such as sodium chloride, potassium chloride, etc.), cocoa, chocolate, cinnamon, vanilla, nutmeg, coconut, almond, combinations thereof, and the like. The dessert composition may contain a sweetening agent in an amount between 1% and 20%, between 1.5% and 10%, between 5% and 20%, or between 2% and 18%. The dessert composition may contain no sweetening agent. The dessert composition may contain a salt in an amount between 0.001% and 3.0%, between 0.01% and 2.0%, or between 0.025% and 1.75%. The dessert composition may contain no salt.

[0090] The dessert composition may contain a sweetening agent. Suitable sweetening agents are known and described in the art. The sweetening agent may be at least one of a calorie-free sweetening agent or a calorie-containing sweetening agent. The sweetening agent may be any type of sweetening agent, for example, a sweetening agent obtained from plants or plant products, or a physically modified or chemically modified sweetening agent obtained from plants, or a synthetic sweetening agent. Exemplary sweetening agents include steviol glycosides, mogrosides, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, tagatose, trehalose, galactose, rhamnose, cyclodextrins (such as α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin), ribulose, threose, arabinose, xylose, lyxose, allose, altrose, mannose, idose, lactose, maltose, invert sugar, isotrehalose, neotrehalose, palatinose or isomaltulose, erythrose, deoxyribose, gulose, idose, talose, erythrulose, xylulose, allulose, turanose, cellobiose, glucosamine, mannosamine, fucose, fucoidan, glucuronic acid, gluconic acid, glucono-δ-lactone, abequose, galactosamine, xylo-oligosaccharides (such as xylotriose, xylobiose, etc.), gentiobiose-oligosaccharides (such as gentiobiose, gentiotriose, gentiotetraose, etc.), galacto-oligosaccharides, sorbose, dihydroxyacetone, glyceraldehyde, Aspergillus niger oligosaccharides, fructo-oligosaccharides (such as kestose, nystose, etc.), maltotetraose, maltotriitol, tetrasaccharides, manno-oligosaccharides, malto-oligosaccharides (such as maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, etc.), dextrin, lactulose, melibiose, raffinose, rhamnose, ribose, sucralose, acesulfame K, aspartame, saccharin, conjugated sugars, soybean oligosaccharides, and combinations thereof. When applicable, the D-configuration or L-configuration may be used.

[0091] The dessert composition may contain an acid. Suitable acids include, but are not limited to, citric acid, lactic acid, sorbic acid, malic acid, combinations thereof, and the like. The dessert composition may contain an acid in an amount of up to 0.001%, up to 0.005%, up to 0.01%, up to 0.1%, up to 1.0%, up to 1.5%, or up to 2.0% of the dessert composition. The dessert composition may contain an acid between 0.0001% by weight and 2.0% by weight, between 0.0002% by weight and 1.5% by weight, between 0.0003% by weight and 1.0% by weight.

[0092] Example

[0093] The present invention is further described in detail by reference to the following experimental examples. Unless otherwise specified, these examples are provided for illustrative purposes only and are not intended to be limiting. Accordingly, the present invention should in no way be construed as limited to the following examples, but should be construed to cover any and all variations that become apparent as a result of the teachings provided herein.

[0094] Example 1 – Fermented pea protein with Leuconostoc mesenteroides

[0095] 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.

[0096] 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 h 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 min. The syneresis observed in the control starch and protein suspensions compared to the absence of syneresis observed in Sample 1.2 is shown in Figure 8 in. Appearance was evaluated visually.

[0097] 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 into their own mouths, dispersed the sample around their mouths, and recorded their observations. Visual evaluations were also made of the texture and appearance observations.

[0098] Photographs of the samples outlined in Table 1 before and after manual agitation for approximately 2 s to 5 s are provided in Figure 1 、 Figure 2 and Figure 3 in.

[0099] Overall, despite some strain-to-strain variability in the fermented samples, all strains showed significant viscosity effects, 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 was a starter dough with a very smooth, shiny, and glistening texture, rich in acidic and umami notes. Leuconostoc citreum C22B11 started to thicken faster (around 4 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 had a very viscous and slippery texture, similar to melted cheese (see Figure 4 ).

[0100] Table 1 .

[0101]

[0102]

[0103] Example 2 - Viscosity and acidification

[0104] The viscosity of the samples outlined in Table 1 was tested together with a blank pea protein sample containing pea protein isolate, sucrose, and water but not inoculated with any Leuconostoc strains. Viscosity (cP) was measured using a Rapid Visco Analyzer (RVA) at 25 °C and 250 rpm. As Figures 5 to 7 and shown in 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.

[0105] 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 no change or loss in thickness after freezing and thawing.

[0106] 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 pronounced for strain B3K7. 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.

[0107] Table 2 .

[0108]

[0109]

[0110] Example 3 – Composition analysis

[0111] 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 fermentate. 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.

[0112] Table 3 .

[0113]

[0114]

[0115] Example 4 – Pea protein dessert

[0116] 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 (approx. 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.

[0117] 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.

[0118] 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 .

[0119] Table 4 .

[0120] Ingredient Weight % Pea protein isolate 4.0 Sucrose 7.0 Hydroxypropylated starch 5.0 Carrageenan 0.015 Water 84.0

[0121] Table 5 .

[0122] 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

[0123] Example 5 – Comparative pea protein fermentation

[0124] Fermentation of pea protein by three Leuconostoc citreum strains and one Leuconostoc pseudomesenteroides strain was compared to fermentation by a dairy-isolated lactic acid bacteria culture containing Streptococcus thermophilus and Lactobacillus bulgaricus (trade name “YO-MIX 433”). 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.

[0125] 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.

[0126] 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 .

[0127] Generally, compared with the YO-MIX 433 culture, the strains C18X1, B3K7, C22B11, and C18X24 produce pea protein fermentates with higher viscosities. Under the experimental conditions of Example 1 and the experimental conditions of US2020 / 296982, the strains C18X1, B3K7, C22B11, and C18X24 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 strains C18X1, B3K7, C22B11, and C18X24 again show products with higher viscosities compared to the YO-MIX 433 culture. Additionally, the products fermented with the strains C18X1, B3K7, C22B11, and C18X24 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.

[0128] Table 6 .

[0129]

[0130] Table 7 .

[0131]

[0132]

[0133] Table 8 .

[0134]

[0135] Table 9 .

[0136]

[0137]

[0138] Example 6 – Comparative plant protein fermentation

[0139] 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.

[0140] Table 10 .

[0141] 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

[0142] Table 11 .

[0143]

[0144]

[0145] Terms describing the present invention

[0146] Clause 1. A vegan dessert composition, the vegan dessert composition comprising:

[0147] A fermented pea protein composition; wherein the fermented pea protein composition is made by contacting pea protein with Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) or Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) in the presence of sucrose; and

[0148] Optionally, a sweetener, a flavorant, or a combination thereof.

[0149] Clause 2. The composition according to Clause 1, wherein the vegan dessert composition does not contain added starch.

[0150] Clause 3. The composition according to Clause 1 or Clause 2, wherein the vegan dessert composition comprises a sweetener.

[0151] Clause 4. The composition according to Clause 3, wherein the sweetener comprises steviol glycosides, mogrosides, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

[0152] Clause 5. A method for producing a vegan dessert composition, the method comprising:

[0153] (i) contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum B3K7

[0154] (BCCM accession number LMG P - 32801) or Leuconostoc citreum C22B11 (BCCM accession number LMG P - 32800) for a period of time under conditions sufficient to produce a fermented pea protein product;

[0155] (ii) optionally, inactivating the Leuconostoc citreum bacteria in the fermented pea protein (e.g., by pasteurization, heat killing, irradiation, or chemical treatment); and

[0156] (iii) adding a sweetener, a flavorant, or a combination thereof to the fermented pea protein product,

[0157] thereby producing a vegan pudding composition.

[0158] Clause 6. The method according to Clause 5, wherein the pea protein 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.

[0159] Clause 7. The method according to Clause 5 or 6, wherein the pea protein composition is fermented for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; and / or the pea protein 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.

[0160] Clause 8. The method according to any one of Clauses 5 to 7, the method further comprising a step of stirring the contacted pea protein composition during or after step (i).

[0161] Clause 9. The method according to any one of Clauses 5 to 8, wherein the sweetener comprises stevioside, mogroside, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

[0162] Clause 10. A composition comprising Leuconostoc citreum B3K7 (BCCM accession number LMG P - 32801), sucrose, and pea protein.

[0163] Clause 11. Isolated Leuconostoc citreum B3K7 (BCCM accession number LMG P - 32801) bacterial cells.

[0164] Use of Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) for the production of a vegan dessert composition.

[0165] Clause 13. A composition comprising Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), sucrose, and pea protein.

[0166] Clause 14. Isolated bacterial cells of Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800).

[0167] Use of Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) for the production of a vegan dessert composition.

[0168] Clause 16. A composition comprising:

[0169] Fermented pea protein; and

[0170] Inactive bacterial cells of Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) and / or Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800);

[0171] wherein the composition does not contain added sucrose and / or wherein the composition does not contain added starch.

Claims

1. A vegan dessert composition, the vegan dessert composition comprising: A fermented pea protein composition, wherein the fermented pea protein composition is made by contacting pea protein with Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) or Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) in the presence of sucrose; and Optionally, a sweetener, a flavorant, or a combination thereof.

2. The composition according to claim 1, wherein the vegan dessert composition does not contain added starch.

3. The composition according to claim 1 or claim 2, wherein the vegan dessert composition comprises a sweetener.

4. The composition according to claim 3, wherein the sweetener comprises stevioside, mogroside, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

5. A method for producing a vegan dessert composition, the method comprising: (i) Contacting a pea protein composition comprising pea protein and sucrose with Leuconostoc citreum B3K7 (BCCM accession number LMGP-32801) or Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) for a period of time under conditions sufficient to produce a fermented pea protein product; (ii) Optionally, inactivating the Leuconostoc citreum bacteria in the fermented pea protein (e.g., by pasteurization, heat killing, irradiation, or chemical treatment); and (ii) Adding a sweetener, a flavorant, or a combination thereof to the fermented pea protein product, thereby producing a vegan pudding composition.

6. The method according to claim 5, wherein the pea protein 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.

7. The method according to claim 5 or 6, wherein the pea protein composition is fermented for at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours; and / or The pea protein 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.

8. The method according to any one of claims 5 to 7, the method further comprising a step of stirring the contacted pea protein composition during or after step (i).

9. The method according to any one of claims 5 to 8, wherein the sweetener comprises stevioside, mogroside, sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, sucralose, acesulfame potassium, aspartame, saccharin, or a combination thereof.

10. A composition, said composition comprising Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801), sucrose and pea protein.

11. Isolated Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) bacterial cells.

12. Use of Leuconostoc citreum B3K7 (BCCM accession number LMG P-32801) for the production of a vegan dessert composition.

13. A composition, said composition comprising Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800), sucrose and pea protein.

14. Isolated Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) bacterial cells.

15. Use of Leuconostoc citreum C22B11 (BCCM accession number LMG P-32800) for the production of a vegan dessert composition.

Images