Sweet umami plant-protein product and process of producing the same and uses thereof

Abstract

The present invention relates to a process for producing sweet umami plant- proteins, as well as fermented food or feed containing said plant-proteins, offering a sweet-umami sensory profile, improved nutritional value, and with multiple textural functionalities. Specifically, the invention involves a process of fermenting plant-proteins through solid-state fermentation (SSF), enzymatic treatment and submerged fermentation (SMF). The invention also encompasses the resulting fermented products having sweet-umami- sensory profile, improved nutritional value and beneficial texture attributes in various product use cases as well as various applications thereof. Even further, the invention also related to sweet tasting plant-protein product produced by fermentation, wherein the plant-protein product has umami, volatile aroma compounds of honey and / or caramel, and polyalcohols such as erythritol, mannitol and glycerol, and the plant-protein product comprises at least 30 % protein.

Description

[0001] P85424PC01

[0002] Sweet umami plant-protein product and process of producing the same and uses thereof

[0003] Technical field of the invention

[0004] The present invention relates to a process for producing sweet umami plantproteins, as well as fermented food or feed containing said plant-proteins, offering a sweet-umami sensory profile, improved nutritional value, and with multiple textural functionalities. Specifically, the invention involves a process of fermenting plant-proteins through solid-state fermentation (SSF), enzymatic treatment and submerged fermentation (SMF). The invention also encompasses the resulting fermented products having sweet-umami sensory profile, improved nutritional value and beneficial texture attributes in various product use cases as well as various applications thereof. The invention furthermore relates to a sweet tasting plant-protein product produced by fermentation, wherein the plant-protein product has umami and volatile aroma compounds that contribute to honey, fruity and / or caramel notes and the plant-protein product comprises at least 30 % protein, whereby the resulting plant-protein product ingredient can replace up to 60% sugar in food / feed applications without comprising the sensory satisfaction.

[0005] Background of the invention

[0006] Sweetness is deeply ingrained in human experience, both biologically and psychologically. Carbohydrates, naturally linked to sweetness, are essential energy sources, and the taste of sweetness evokes powerful hedonic responses. These positive emotions associated with sweet tastes are not just a product of individual preference but a result of complex biological processes. Sweet taste receptors on the tongue detect sugars, which trigger a cascade of neural events that stimulate the brain's reward system, creating a pleasurable sensation (Beauchamp, 2016). This response is evolutionarily conserved, meaning that it has persisted across species as a beneficial mechanism to encourage the consumption of energy-dense foods.P85424PC01

[0007] Sugars, such as glucose, sucrose, fructose, maltose, and lactose, are particularly potent in inducing sweetness and are crucial sources of glucose, the brain's primary metabolic fuel. The brain relies heavily on glucose for optimal functioning, and the consumption of sweet foods can create strong motivation due to the direct and pleasurable link between taste and energy provision. However, this natural drive for sweetness, while beneficial in evolutionary contexts of scarcity, becomes problematic in modern environments where sugars are highly available and frequently consumed in excess (von Molitor et al., 2021).

[0008] Recommendations from health organizations, like the World Health Organization (WHO), suggest reducing free sugar intake to less than 10 % of daily energy and further to 5 % for greater health benefits. However, many populations, particularly younger groups, exceed these guidelines (Gillespie et al., 2023a).

[0009] While sugars are essential for energy, excessive consumption has increasingly become a global health issue. Diets high in sugar, especially added sugars found in processed foods and beverages, are linked to a range of serious health conditions. The overconsumption of sugar is a major contributing factor to obesity, which affects about 13 % of the global population. Obesity, in turn, increases the risk of developing noncommunicable diseases (NCDs) such as type 2 diabetes, cardiovascular disease, and certain cancers (Gillespie et al., 2023b). These NCDs account for approximately 74 % of global deaths annually, with 85 % of premature deaths occurring in low-and middle-income countries (LMICs) (World Health Organization, 2023).

[0010] While sweetness plays a vital role in human biology by providing energy and triggering positive emotional responses, the modern prevalence of high sugar consumption poses significant health risks. This imbalance between evolutionary biology and contemporary dietary habits has led to a global rise in preventable, sugar-related diseases.

[0011] Additionally, the connection between sweetness and umami involves the roles of T1R taste receptors, specifically the T1R1 / T1R3 heterodimer for umami and the T1R2 / T1R3 heterodimer for sweetness. These receptors, part of the TAS1R family,P85424PC01

[0012] allow animals to detect a variety of flavors - amino acids for umami and sugars for sweetness - through related but distinct signalling mechanisms. This functional similarity within the TAS1R family illustrates an evolutionary adaptation, enabling animals to recognize essential nutrients and energy sources, thereby linking the perception of sweetness and umami in sensory experience.

[0013] Umami compounds, such as monosodium glutamate (MSG) and specific gammaglutamyl peptides like glutamate-glutamate (Glu-Glu) and glutamate-aspartate (Glu-Asp), interact with sweet taste receptors (T1R2 / T1R3). Rather than directly enhancing sweetness, this interaction primarily modulates sweetness perception. When these umami compounds are co-applied with sweet substances like sucrose, they inhibit the activation of sweet taste receptors, particularly those sweeteners that bind to the extracellular domain of T1R2. This suggests that umami compounds may alter sweetness perception allosterically, especially when combined with certain sweeteners (Shim et al., 2015).

[0014] US20220295846A1 describes a process involving the enzymatic hydrolysis of rice protein, followed by the separation and recovery of the supernatant. The resulting ingredient enhances sweetness, improves mouthfeel, and masks off-notes in food and beverages. Additionally, it helps inhibit the degradation of sweeteners, improving taste retention and enabling lower-calorie formulations.

[0015] WO2015 / 012465A1 describes a process using Aspergillus spp. and Corynebacterium spp. in a two-step fermentation process. The method utilizes grains such as corn, wheat, soybean, rice, and wheat gluten, along with glutamic acid and inosine-5'-monophosphate (IMP), to create a kokumi flavor. After fermentation, the reaction is followed by activated carbon treatment and filtration to refine the flavor, enhancing the richness and mouthfeel of food products naturally.

[0016] EP3259998B1 describes flavor compositions containing tripeptides, including pyroglutamic acid and specific amino acids such as valine, leucine, and cysteine. These compositions enhance or modify the taste, flavor, and mouthfeel of various food products by increasing saltiness or umami intensity. The invention aims to provide clean saltiness and umami taste at low concentrations, reducing the need for sodium chloride and enhancing flavor in both sweet and savory foods.P85424PC01

[0017] However, any known processes or ingredients provide either sweetness or umami, not combination of both sweet and umami while also supporting high protein content claim (>20 %).

[0018] Hence, a new and improved process for producing sweet umami plant-proteins, as well as fermented food or feed containing said plant-proteins, offering a sweetumami sensory profile, improved nutritional value, and with multiple textural functionalities, would be advantageous.

[0019] Despite growing advances in sweetness modulation, kokumi enhancement, and umami-rich ingredients, current solutions remain fundamentally fragmented. Existing approaches typically target either sweetness perception (e.g., enzymatic hydrolysates or sweetness enhancers) or umami / kokumi enhancement (e.g., glutamate-rich fermentations or peptide-based flavor modulators), but rarely both in an integrated manner. Moreover, many of these solutions rely on low-protein matrices, refined additives, or flavor-active fractions used at trace levels, which limits their nutritional contribution and constrains their application in products positioned for high-protein claims (>20 %). As a result, manufacturers are often forced to combine multiple ingredients— sweeteners, umami enhancers, texturizers, and protein isolates— leading to complex formulations, compromised clean-label positioning, suboptimal sensory coherence, and reduced consumer satisfaction due to mismatched taste, mouthfeel, and nutritional expectations.

[0020] The process of the invention addresses these drawbacks by enabling the production of intrinsically sweet-umami plant-protein ingredients in which flavor, nutrition, and functionality are co-developed rather than combined at the formulation stage. By integrating fermentation-driven flavor formation with protein-rich plant substrates, the approach aligns sweetness modulation, umami depth, and kokumi-like mouthfeel within a single biomass-based ingredient. This creates a synergistic sensory effect that enhances palatability while supporting high protein content, improved nutritional quality, and multiple textural functionalities. Importantly, this strategy responds directly to contemporary challenges in sugar reduction, clean-label formulation, and protein -forward foods, offering a biologically grounded and technologically scalable pathway to improveP85424PC01

[0021] consumer satisfaction without relying on excessive sugars, artificial sweeteners, or isolated flavor additives.

[0022] Summary of the invention

[0023] An object of the present invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows: (a) in the solid-state fermentation (SSF) step; heating of a soaked and rinsed grain and / or legume substrate rich (above 20 % w / w) in protein and sweet amino acids, followed by; cooling and subsequent inoculation with Aspergillus spp., followed by; a first incubation thereby obtaining koji, followed by; mixing said koji with grain or legume rich in protein and amino acids and water, followed by; a second incubation, followed by; the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either: directly after cooling or in dry powder form after drying (b) in the enzymatic treatment step; mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by; adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by; inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by; the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either: directly after cooling or in dry powder form after drying; (c) in the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by; inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; second fermentation with non-alcoholic or alcoholic yeast under nonstatic conditions, followed by pasteurization, followed by; drying to obtain a sweet-umami plant protein product.

[0024] Another aspect of the present invention is to provide a food product comprising a fermented sweet-umami plant-protein ingredient containing at least 30% protein and providing sweet and umami taste together with fruity, honey, caramel, nutty, and / or roasted notes and further comprising one or more polyalcohols selected from erythritol, mannitol, and glycerol, wherein the food product (i) shows sameP85424PC01

[0025] or better flavor characteristics compared to a control product without the sweetumami plant-protein ingredient and / or (ii) provides increased mouthfeel at an equivalent sweet-taste level without requiring higher sugar content and / or (iii) masks the off / bitter taste of plant-protein.

[0026] Brief description of the figures

[0027] Figure 1 shows an overview of the cross-fermentation process with one submerged fermentation (SMF) step according to the present invention.

[0028] Figure 2 shows an overview of the cross-fermentation process with two submerged fermentation (SMF) steps including B12 and sugar alcohols production according to another aspect of the invention.

[0029] Figure 3 shows aroma compounds, esters, alcohols, aldehydes and ketones, in samples fermenting with yeasts, Saccharomyces pastorianus, Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans, following the process shown in figure 1 (1st submerged fermentation).

[0030] Figure 4 shows polyalcohols, such as erythritol, maltitol, threitol, d-arabitol, and glycerol, before and after fermentation with Saccharomyces pastorianus, at 25 °C, for 16 h, non-static anaerobic fermentation, following the process shown in figure 1 (1st submerged fermentation).

[0031] Figure 5 shows amino acids concentration raw material (without fermentation) and sweet umami plant protein (final ingredient) with fermentation with Saccharomyces pastorianus at 25 °C, for 16 h, non-static and anaerobic fermentation, following the process shown in figure 1 (1st submerged fermentation).

[0032] Figure 6 shows glutamic acids concentration raw material (without fermentation) and sweet umami plant protein (final ingredient) with fermentation with Saccharomyces pastoruanus at 25 °C, for 16 h, non-static and anaerobic fermentation, following the process shown in figure 1 (1st submerged

[0033] fermentation).P85424PC01

[0034] Figure 7 shows gamma-glutamyl peptides responsible for kokumi, formed from bitter amino acids through glutamylation, whereby a gamma-glutamyl (y-Glu) group from a donor molecule (such as glutamine) is attached to bitter amino acids including valine, leucine, and phenylalanine. Raw material (without fermentation) and sweet-umami plant protein (final ingredient) after fermentation Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions) were obtained following the process shown in Figure 1 (1stsubmerged fermentation).

[0035] Figure 8 shows all the gamma-glutamyl peptides responsible for kokumi. Raw material (without fermentation) and sweet-umami plant protein (final ingredient) after fermentation with Saccharomyces pastorianus at 25 °C for 16 h under nonstatic and anaerobic conditions were obtained following the process shown in Figure 1 (1stsubmerged fermentation).

[0036] Figure 9 shows all the vitamins (vitamins Bl, B2, B5 and B7) produced during the fermentation process. Raw material (without fermentation) and sweet-umami plant protein (final ingredient) after fermentation with Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions, were obtained following the process shown in Figure 1 (1stsubmerged fermentation).

[0037] Figure 10 A) and B) show all the postbiotic compounds before and after fermentation. Raw material (without fermentation) and sweet-umami plant protein (final ingredient) after fermentation with Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions, were obtained following the process shown in Figure 1 (1stsubmerged fermentation).

[0038] Figure 11 shows the prebiotic compounds analyzed before and after fermentation. Raw material (without fermentation) and sweet-umami plant protein (final ingredient) after fermentation with Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions) were obtained following the process shown in Figure 1 (1stsubmerged fermentation).P85424PC01

[0039] Figure 12 shows butter cookies which were used as a model for color comparison. The cookies were made with sugar, flour, egg, and butter, mixed together, and baked for 15 minutes at 180 °C and containing sweet umami plant-protein product obtained by the process according to the invention: from left to right 171 (control), 513 (20 % sugar reduction without egg), 715 (20 % sugar reduction with egg), 357 (35 % sugar reduction without egg) and 289 (35 % sugar reduction with egg). Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0040] Figure 13 shows a contingency table of all attributes in the consumer sensory test. Y-axis shows the number of times consumers choose a certain attribute. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0041] Figure 14 shows principal Component Analysis (PCA) biplot. Note: Proportion of variance explained by PCI (Principal Component 1) is 52.35 %, and by PC2 (Principal Component 2) is 35.07 %. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0042] Figure 15 shows mean intensity of descriptors in the consumer sensory analysis. Y-axis shows an intensity scale from 1-5 points. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0043] Figure 16 shows mean liking of taste, texture and aroma in the consumer sensory analysis. Y-axis shows a liking scale from 1-9 points. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).P85424PC01

[0044] Figure 17 shows the liking score of texture of samples 357, 715, 289 and 513 compared to control 171. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0045] Figure 18 shows the overall of texture liking in consumer sensory analysis of samples 357, 715, 289 and 513 compared to control 171. Y-axis liking scale from 1-9 points. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus at 25 °C for 16 h under non-static and anaerobic conditions).

[0046] Figure 19 shows the intensity scale for umami, bitter and sweet taste consumer sensory analysis of protein bar samples, 69, 71 and 72, compared to control sample without sweet umami plant protein. Y-axis intensity scale from 1-15. Sweet umami plant protein was fermented following the shown figure 1 (1stsubmerged fermentation and Saccharomyces pastorianus, Pichia kluyveri and Torulaspora delbrueckii at 25 °C for 16 h under non-static and anaerobic conditions.

[0047] Figure 20 shows the hedonic scale for liking sensory analysis of protein bar samples 69, 71 and 72, compared to control sample without sweet umami plant protein. Y-axis liking scale from 1-9 points.

[0048] The present invention will now be described in more detail in the following.

[0049] Detailed description of the invention

[0050] The first part of the invention relates to solid-state fermentation (SSF) of grains and / or legumes rich in protein and sweet amino acids (i.e. any grains and or legumes with a high concentration of glutamic acid, glycine, alanine, glutamine, and threonine, respectively) for using an Aspergillus spp, such as Aspergillus oryzae and Aspergillus sojae, which converts starch into glucose via saccharification. Most of the carbohydrates are glucose derived from starch, which is broken down by a-amylase and glucoamylase secreted by Aspergillus oryzae. InP85424PC01

[0051] addition to its sweet flavor, it contains glucose, oligosaccharides, amino acids, and vitamins such as Bl, B2, and B6.

[0052] In parallel, enzymatic treatment of proteins derived from protein-rich grains and / or legumes, such as faba beans and / or soy protein and / or oat protein, produces sweet amino acids such as alanine, glutamine, glycine, serine, threonine, and proline by the action of hydrolases (EC 3.4, including proteases, peptidases). To reduce bitterness, one method involves the y-glutamylization reaction, which synthesizes peptides such as y-Glu-Phe, y-Glu-Leu, or y-Glu-Val (kokumi peptides) from bitter amino acids like Phe, Leu, and Vai. This process is carried out through GGT (gamma-glutamyl transferase) catalysis or any glutaminase (EC 3.5, EC 2.6, and EC 2.3). Additionally, glutaminase enzymes synthesis other y-glutamyl peptides (kokumi peptides), responsible for enhancing sweetness, umami, and reducing bitterness. During fermentation, compounds like glutamate and glutamic acid are released into the medium, enhancing the umami taste and modulating the perception of sweetness. This natural process can contribute to a more balanced flavor profile, helping to reduce the need for added sugars.

[0053] Using one or more species from the group of yeast e.g. Saccharomycodes ludwigii, Pichia kluyveri, Toluraspora delbrueckii, Lanchancea thermotolerans, and other non-Saccharomyces yeasts or Saccharomyces offer unique contributions to the sensory profiles in the product, leading to distinctive and diverse aromatic profiles during the fermentation process.

[0054] Objective of the invention

[0055] Using synergistic cross-fermentation techniques, the inventors have aimed to develop a unique sweet umami plant-protein ingredient with improved nutritional benefits.

[0056] The objective is to reduce overall sugar content by 20-60 % in final products without compromising sweetness perception, while enhancing umami, creaminess, and introducing various aromas to create a more complex flavor profile in the end product.P85424PC01

[0057] In addition, the inventors have improved the nutritional value, including increased protein quality and reduced anti-nutrients in the product produced by the process of the invention.

[0058] The process according to the invention enhances both sweetness and umami through harnessing plant-protein. By utilizing natural carbohydrates, sweet amino acids like glycine and alanine, and aromatic compounds like honey and caramel through fermentation, consumers' perception of sweetness will be enhanced.

[0059] Cross-fermentation increases glutamic acid production to deepen umami, while kokumi peptides (T-glutamyl), e.g. produced by the enzymatic treatment step of the invention, synergistically amplify both sweetness and umami, reducing bitterness for a more complex and appealing flavor profile, and when this is expressed in a combined process the overall sensory experience is uniquely heighten in sweeteners and likeness in comparisons to layering compounds individual.

[0060] That synergistic effect is enhanced by using yeast and bacteria during crossfermentation to produce aromas such as caramel, fruity, and honey-like, and potentially produce sugar alcohol such as mannitol. Nutritional value is improved by increasing protein, essential amino acids, fibre (prebiotic), and postbiotic compounds and e.g. B12. This allows for a significant reduction in sugar and fat without compromising flavor, resulting in a healthier, more flavorful product.

[0061] Additionally, the functional properties of this process that uniquely is combined with sweet sensory and off / bitter masking of plant-protein, can also enhance the texture and stability of the final product. It contributes to reducing sugar while improving hydrocolloid properties, foaming, and emulsifying capacity, due to its water-holding ability, solubility, and pH stability. The presence of natural simple sugars and amino acids makes it ideal for baking, enhancing caramelization and crispiness during the Maillard reaction, thus improving texture and flavor in baked goods.

[0062] Products resulting from the process of the inventionP85424PC01

[0063] Plant-based diets encounter challenges in both taste and nutritional completeness. The unappealing flavor of plant-proteins leads to the incorporation of excessive sugar and additives in non-savory applications like alternative dairy, yogurt, protein bars / powders etc. Apart from that, certain predominantly nutrients, such as B12, Omega 3, etc., in meat or dairy products are scarce in plant-based diets, resulting in potential nutritional deficiencies. Consumers may need an intake of extra food supplements to meet daily nutritional needs.

[0064] The inventor solution aims to create a naturally perceived sweet-umami plantprotein ingredient, with functional attributes and nutritional qualities, and thereby removing barriers to plant-based food consumption and promoting even healthier product formulations.

[0065] Definitions

[0066] Prior to discussing the present invention in further details, the following terms and conventions will first be defined:

[0067] Artificial sweeteners: when used herein artificial sweeteners refer to aspartame, sucralose, saccharin, neotame, acesulfame potassium, and advantame, which are commonly used in sugar-free and low-calorie food and beverages such as diet sodas, sugar-free desserts, chewing gum, and processed foods. They are also found in many "diet" or "light" versions of products, as well as in tabletop sweeteners marketed to consumers looking to reduce sugar intake. Despite being widely considered safe, concerns have arisen about their long-term health effects, particularly on cardiovascular health. Research suggests that regular consumption of these sweeteners may increase the risk of metabolic syndrome, including hypertension, insulin resistance, and abdominal obesity, all linked to cardiovascular disease. Additionally, they can disrupt gut microbiota, potentially leading to metabolic and inflammatory changes, and impair HDL function, contributing to atherosclerosis. Some, like aspartame, have also been associated with arrhythmias and structural heart changes (Singh et al., 2023).

[0068] Cross fermentation: When used herein, cross-fermentation refers to as sequential or parallel fermentation, which originally exits in traditional fermentation practices by the spontaneous occurrence of fungi, bacteria, and yeast during differentP85424PC01

[0069] developmental fermentation stages, for example wine, beer, cheese, sauerkraut, miso, kimchi and coco production, where the utilization of indigenous communities including bacteria and fungi.

[0070] Enzymes: when used herein enzymes may e.g. refer to naturally occurring proteins that act as catalysts to accelerate biochemical reactions. In the food industry, they are sourced from plants, animals, or microorganisms through fermentation and are used to improve manufacturing processes. Enzymes also play a key role in converting starch into ingredients used in various food products, enhancing efficiency and product quality across multiple food processing sectors.

[0071] Grains rich in protein and sweet amino acids: when used herein grains rich in protein and sweet amino acids refer to barley (Hordeum vulgare), oat (Avena sativa), millet (Panicum miliaceum), quinoa (Chenopodium quinoa), corn / maize (Zea mays), rice (Oryza sativa), buckwheat (Fagopyrum esculentum), rye (Secale cereale), wheat (Triticum ssp.), sorghum (Sorghum bicolor), amaranth (Amaranth) , bulgur (Triticum durum), teff (Eragrostis tef), spelt (Triticum spelta), fonio (Digitaria exilis), emmer (hullet wheat), triticale, einkorn wheat (Triticum monococcum) or any type of grains.

[0072] Grain and / or legumes rich in protein and sweet amino acids: when used herein grains and / or legumes rich in protein and sweet amino acids refer to faba (Vicia faba L.), lentil (Vicia lens or Lens culinaris), lupin (Lupine), beans (Phaseolus vulgaris L.), chickpeas (Cicer arietinum L.), peanuts (Arachis hypogaea) or any family Fabaceae(or Leguminosae), or the fruit or seeds of such plants (pulses).

[0073] Koji: When used herein, Koji refers to whole grains or legumes, such as beans that have been inoculated with one or more Aspergillus species, such as Aspergillus oryzae.

[0074] Kokumi peptides: when used herein Kokumi peptides, may e.g. refer to y-glutamyl peptides, being increasingly explored in food science to improve the taste profile of products, offering a fuller, more rounded flavor experience while potentially reducing the need for added sugars, salts, or fats.P85424PC01

[0075] Non-alcoholic yeast: when used herein, non-alcoholic yeast refers to a type of yeast specifically selected or engineered to produce little to no alcohol during fermentation. This is achieved by using yeast strains that do not ferment certain sugars, such as maltose and maltotriose. Examples of such yeast are Saccharomycodes spp., particularly Saccharomycodes ludwigii, which is a yeast species often used in the production of non-alcoholic products and beverages. Although typically associated with alcoholic fermentation, some strains of Saccharomyces cerevisiae, have the ability to produce lower levels of alcohol while still contributing to the desired flavor and aroma profiles. Still another example of non-alcoholic yeasts is Zygosaccharomyces ssp., Pichia ssp. or lactic acid bacteria, such as Streptococcus ssp., Lactococcus ssp., Lactobacillus ssp.

[0076] Plant-protein: when used in the present invention, the term plant-protein shall be understood in the context of the increasing interest in plant-proteins is driven by several key factors, including their potential health benefits, environmental sustainability, and ethical considerations. Consumers are increasingly turning to plant-based diets due to concerns about the adverse health effects associated with high animal protein intake, such as increased saturated fat. Additionally, plant-based proteins are viewed as a more environmentally sustainable option, requiring fewer resources and generating fewer greenhouse gases compared to animal protein production. However, the nutritional quality of plant-proteins is sometimes considered inferior to animal proteins due to lower concentrations of indispensable amino acids, although this can be managed by consuming a variety of plant-protein sources. Plant-proteins have also been associated with various health benefits, such as reducing the risk of cardiovascular disease, managing obesity, and improving metabolic health (Hertzler et al., 2020).

[0077] Solid-state-fermentation (SSF): When used herein, the term solid-state fermentation (SSF) refers to a biomolecule manufacturing process used e.g. in the food industry. These biomolecules are mostly metabolites generated by microorganisms grown on a solid support selected for this purpose. This technology for the culture of microorganisms is an alternative to liquid or submerged fermentation, used predominantly for industrial purposes. Solid state fermentation uses cultures and substrates with low water levels (reduced water activity), which is particularly appropriate for mold.P85424PC01

[0078] Solid fermented grains / legumes: When used herein, Solid fermented grains / legumes refers to the result of the solid-state fermentation (SSF) process using various grains and legumes, with different microorganisms involved.

[0079] Submerged fermentation (SMF): When used herein, the term submerged fermentation (SMF) refers to the substrate being submerged in liquid has conditions that limits oxygen exposure, promoting anaerobic growth of desired microorganisms. Water or brine is a dominant component of the substrate.

[0080] Sugar reduction: when used in the context of the present invention, the term sugar reduction may refer to low sugar, no added sugar, reduced sugar, and sugar-free. Low sugar products contain sugar below a defined limit per serving, while reduced sugar means at least 25-30 % less sugar than the regular version. No added sugar products may still have naturally occurring sugars like lactose or fructose. Sugar-free products contain no added sugars or naturally occurring sugars from ingredients like milk or fruit. "No added sugar" is the top claim on food and beverage launches, followed by "sugar-free" and "low sugar." These claims are especially popular in beverages and confectionery, while categories such as flavored alcoholic drinks, coffee, and desserts are rapidly adopting sugar reduction. Innovations in sweeteners, such as natural substitutes and sweet fibres, help manufacturers replicate sugar's properties like sweetness and moisture retention while reducing calories. Manufacturers are using a variety of sweeteners, both artificial and natural, such as bulk sweeteners like erythritol and natural options like stevia. Innovations in sugar reduction also include the use of fibres like chicory inulin and resistant dextrin, as well as enzymes that convert sugars into fibres (Chen et al., 2022; Innova Market Insights, 2024).

[0081] Sweet amino acids: when used in the context of the present invention, sweet amino acids refers to alanine, glutamine, glycine, serine, threonine, and proline and tryptophan. Sweet amino acids are specific building blocks of protein, like glycine, alanine, serine, glutamine, and threonine, that taste sweet to humans, unlike most amino acids which are bitter or umami; they interact with taste receptors to trigger sweetness, with some, like D-tryptophan, being intensely sweet.P85424PC01

[0082] Sweet proteins: when used in the context of the present invention, sweet proteins may be those derived from honey truffles, thaumatin, and brazzein, which are emerging as sugar-free alternatives with no calories and minimal aftertaste. These proteins are gaining popularity as manufacturers look to overcome taste challenges in sugar reduction, using novel ingredients like sugar-protein combinations or improved stevia for better sweetness and fewer off-flavors.

[0083] Consumer acceptance of sweet proteins is influenced by factors such as perceptions of health and naturalness. Studies suggest that positioning sweet proteins as healthy alternatives to sugar is more effective than presenting them as substitutes for artificial sweeteners. However, a potential downside is that consistently increasing the sweetness of food products may reinforce cravings for sweet flavors, potentially leading to overconsumption. This could perpetuate a cycle where people crave sweetness more frequently, which may undermine the long-term goal of reducing sugar intake. Despite the potential for sweet proteins to reduce sugar consumption and address global concerns like obesity and diabetes, there is a risk that constantly enhancing sweetness could negatively impact consumer behaviour by encouraging habitual reliance on sweet-tasting foods (Banovic & Grunert, 2024). Additionally, advances in natural sweeteners, fibres, and protein-based alternatives are set to transform the sugar reduction landscape, offering healthier options without compromising taste. High-intensity and artificial sweeteners can negatively affect the consumer experience by distorting taste expectations and potentially increasing sugar tolerance, leading to cravings for more sweet products. Kokumi y-glutamyl peptides offer a potential solution by enhancing the perception of sweetness, thereby helping to reduce sugar consumption (Wang et al., 2022). These peptides interact synergistically with basic tastes, such as salty, sweet, and umami, intensifying their flavor without contributing a distinct taste of their own (Wang et al., 2022). Additionally, umami compounds like glutamic acid, glutamate, and inosine 5'-monophosphate (IMP) have been shown to modulate sweet taste, further supporting sugar reduction efforts (Shim et al., 2015).

[0084] Sweet-tasting proteins: when used in the context of the present invention, sweettasting proteins may refer to proteins, such as thaumatin, mabinlin, monellin, pentadin, brazzein, miraculin, curculin, and lysozyme, which have gainedP85424PC01

[0085] recognition fortheir potential as natural sweeteners. Except for lysozyme, which is derived from egg whites, all of these proteins are originally expressed and isolated from tropical plants. Thaumatin (E957) is the most extensively studied, regulated, and commercially available sweet protein. Among these, brazzein production produced via precision fermentation has shown a promising result, particularly when expressed in Pichia pastoris, which has proven to be highly efficient in producing functional brazzein at high yields. Media used in precision fermentation contains multiple carbon sources such as glucose, mannose, galactose, and fucose moieties derived from sugarcane, corn, or sugar beet. In order to ensure proper protein folding and functionality, the transglycosylase activity is highly controlled, which involves a follow-up complex and energy consuming purification process. Advantages associated with sweet-tasting proteins lies e.g. in the ability to synthesize these proteins through fermentation enabling large-scale production of sweet proteins. Some of the disadvantages concerning e.g. recombinant sweet proteins are regulatory challenges, particularly in the European Union, where their use in food products is restricted.

[0086] Umami: When used herein, the term umami refers to one of the five basic tastes, along with sour, sweet, bitter, and salty, and comes from three compounds that are naturally found in plants and meat: glutamate, inosinate, and guanylate. Glutamate is an amino acid found in vegetables and meat. Inosinate is primarily found in meat, and guanylate levels are the highest in plants.

[0087] Sweet amino acids: Sweet amino acids are specific building blocks of protein, like glycine, alanine, serine, glutamine, and threonine, that taste sweet to humans, unlike most amino acids which are bitter or umami; they interact with taste receptors to trigger sweetness, with some, like D-tryptophan, being intensely sweet.

[0088] Embodiments of the invention

[0089] (the process according to Figure 1)

[0090] A first embodiment of the invention relates to the following process steps; solid-state fermentation (SSF), enzymatic treatment, submerged fermentation (SMF) and downstream process, wherein said steps can be carried out under the following conditions:P85424PC01

[0091] Solid-state fermentation (SSF) step

[0092] Soaking a grain and / or legume substrate rich in protein and sweet amino acids, i.e. with a high concentration of glutamic acid, such as oats (could also be maize, rice, sorghum, rye, wheat, barley, pearl millet, faba, wheat bran, BSG, rapeseed / seedpress cake) for at least 0.5-10 h (preferably 1-8 h, more preferably 2-7 h, most preferably 5 h) at 4-30 °C (preferably 20-26 °C, more preferably 20-23 °C, most preferably 22 °C), and rinse with cold water, followed by;

[0093] Heating (e.g. cooking) in e.g. an oven at 60-170 °C (preferably 60-130 °C, more preferably 90-110 °C, most preferably 100 °C) for 0.1-3 h (preferably 0.1-1 h, more preferably 0.2-0.5 h, most preferably 0.2 h), followed by;

[0094] Cooling to 10-35 °C (preferably 15-35 °C, more preferably 30-35 °C, most preferably 32 °C), followed by;

[0095] Inoculate with fungi such as Aspergillus ssp. to 0.01 to 3 g / kg (preferably 0.1-1 g / kg, more preferably 0.2-0.6 g / kg, most preferably 0.3 g / kg), followed by;

[0096] First incubation at 25-45 °C (preferably 25-40 °C, more preferably 30-35 °C, most preferably 32 °C) for 0.5-48 h (preferably 10-48 h, more preferably 24-42 h, most preferably 40 h) and 40-100 % of humidity (preferably 50-80 % of humidity, more preferably 50-70 % of humidity, most preferably 60% of humidity) thereby producing koji, followed by;

[0097] Mixing a ratio of from 0.1:1:0.1 to 1:3:1 of grain or legume rich in protein and amino acids (such as oat): koji: water (preferably 0.5:1:0.1-1:3:1, more preferably 0.5:1:0.5-l:2:l, most preferably 1:2:1), followed by;

[0098] Second incubation, e.g. in an oven at 10-80 °C (preferably 20-60 °C, more preferably 30-50 °C, most preferably 55 °C) for 0.5-18 h (preferably 1-12 h, more preferably 5-10 h, most preferably 8 h), use directly or followed by; a drying step e.g. direct heat drying or freeze drying.

[0099] The resulting slurry / powder product is subsequently subjected to submerged fermentation (see below).

[0100] Enzymatic treatment step

[0101] Mixing protein derived from protein-rich grains and / or legumes, such as faba protein and / or soy protein and / or oat protein (preferably 1-20 % (w / w), more preferably 3-15 % (w / w), most preferably 7 % (w / w)), substrate rich in fibre, such as oat bran (preferably 0.1-5 % (w / w), more preferably 0.5-4 % (w / w), most preferably 3 % (w / w)), and water, (such as 10-80 % (w / w), preferably 40-P85424PC01

[0102] 80 % (w / w), more preferably 50-70 % (w / w), most preferably 63 % (w / w)), mix everything together in a mixer, followed by;

[0103] Adding enzymes in amounts of 0.1-4 g / kg (preferably 0.1-2 g / kg, more preferably 0.1-0.5 g / kg, most preferably 0.2 g / kg) and adjust the pH in a range of 2-9 (preferably 3-8, more preferably 4-6, most preferably 5), at 5-80 °C (preferably 20-60 °C, more preferably 30-60 °C, most preferably 37 °C) for 0.1-24 h (preferably 3-12 h, more preferably 4-10 h, most preferably 5 h) including mixing for 2 h for proteases, 1 h for peptidase and 2 h for glutaminase, wherein the enzymes are:

[0104] o One or more proteases, such as Proteases P "Amano" 6SD, Proteases HF "Amano" 150SD, and ProteAX (EC 3.4.21) o One or more peptidases, such as Peptidase R (including any EC.

[0105] 3.2.1)

[0106] o One or more glutaminases, such as EC 3.5.1,

[0107] followed by;

[0108] Inactivating the enzymes by heat treatment at 90-150 °C (preferably 90-105 °C, more preferably 90-100 °C, most preferably 95 °C) for 1-120 min (preferably 10-60 min, more preferably 10-30min, most preferably 15 min), used directly by cooling the sample before inoculation or followed by a direct heat drying or freeze drying step.

[0109] The resulting powder or slurry product is subsequently subjected to submerged fermentation as follows:

[0110] Submerged fermentation (SMF) step

[0111] Mixing the enzymatically treated protein product resulting from the enzymatic treatment step in amounts of 0.1-50 % (w / w) (preferably 2-40 % (w / w), more preferably 3-10 % (w / w), most preferably 7 % (w / w)), solid fermented grains / legumes in amounts 0.1-50 % (w / w) (preferably 8-40 % (w / w), more preferably 10-30 % (w / w), most preferably 28.5 % (w / w)), followed by;

[0112] Inoculating with one or more alcoholic or non-alcoholic yeasts, such as Saccharomycodes ssp., Saccharomyces spp., Pichia ssp., Toluraspora ssp., Lachancea ssp. or Zygosaccharomyces ssp. in amounts of 0.1-10 % (w / w) (preferably 0.2-5 % (w / w), more preferably 0.5-2 % (w / w), most preferably 1.5 % (w / w)), followed by;P85424PC01

[0113] Fermenting (1ststep SMF) for 2-96 h (preferably 10-60 h, more preferably 12-30 h, most preferably 16 h) at 1-55 °C (preferably 5-40 °C, more preferably 20-30 °C, most preferably 25 °C), under static or non-static and anaerobic or aerobic conditions, followed by;

[0114] Second inoculation (optional) with one or more alcoholic or non-alcoholic yeasts, such as Saccharomycodes ssp., Saccharomyces spp., Pichia ssp., Toluraspora ssp., Lachancea ssp. or Zygosaccharomyces ssp. in amounts of 0.1-10 % (w / w) (preferably 0.2-5 % (w / w), more preferably 0.5-2 % (w / w), most preferably 1.5 % (w / w)), followed by;

[0115] Fermenting (2ndstep SMF) for 2-96 h (preferably 3-60 h, more preferably 4-3 h, most preferably 4 h) at 1-55 °C (preferably 5-40 °C, more preferably 20-30 °C, most preferably 25 °C), under static or non-static and anaerobic or aerobic conditions, followed by;

[0116] Pasteurizing at 60-140 °C (preferably 70-130 °C, more preferably 90-120 °C, most preferably 100 °C) for 0.01-60 min (preferably 5-45 min, more preferably 15-30 min, most preferably 21 min), followed by a cooling step and a downstream process, e.g. drum drying, spray drying or freeze drying.

[0117] A second embodiment of the invention (process with three SMF steps to produce B12, possible sugar alcohol according to figure 2) relates essentially to the same process steps as in the first embodiment relating to figure 1 except that bacteria and yeast-fermentation may optionally be included in the first SMF-step, as follows (see figure 2):

[0118] Mixing the enzymatically treated protein derived from protein-rich grains and / or legumes, such as faba protein (0.1-95 % (w / w)) with solid fermented grains / legumes (0.1-50 % (w / w)), followed by;

[0119] Inoculating with one or more lactic acid bacteria (such as Streptococcus ssp., Lactococcus ssp., Lactobacillus ssp.), Torula spp., Candida magnoliae, Yarrowia lipolytica, and Propionibacterium spp. (0.1-10 %), followed by;

[0120] Fermenting (1ststep SMF) for 2-48 h at 1-50 °C (most preferably at 29 °C) under static or non-static and anaerobic or aerobic conditions, followed by;

[0121] Inoculating with one or more Saccharomycodes ssp., Saccharomyces spp., Pichia ssp., Toluraspora ssp., Lachancea ssp. or Zygosaccharomyces ssp. (0.1-10 %), followed by;P85424PC01

[0122] Fermenting (2ndstep SMF) for 0.1-48 h at 1-50 °C (most preferably at 25 °C) under anaerobic or anaerobic and static or non-static conditions, followed by Third inoculation (optional) with one or more alcoholic or non-alcoholic yeasts, such Saccharomycodes ssp., Saccharomyces spp., Pichia ssp., Toluraspora ssp., Lachancea ssp. or Zygosaccharomyces ssp. in amounts of 0.1-10 % (w / w) (preferably 0.2-5 % (w / w), more preferably 0.5-2 % (w / w), most preferably 1.5 % (w / w)), followed by;

[0123] Fermenting (3rdstep SMF) for 2-96 h (preferably 3-60 h, more preferably 4-12 h, most preferably 4 h) at 1-55 °C (preferably 5-40 °C, more preferably 20-30 °C, most preferably 25 °C), under static or non-static and anaerobic or aerobic conditions, followed by;

[0124] Pasteurizing at 60-140 °C (preferably 70-130 °C, more preferably 90- 120 °C, most preferably 100 °C) for 0.01-60 min (preferably 5-45 min, more preferably 15-30 min, most preferably 21 min), followed by a cooling step and a downstream process, e.g. drum drying, spray drying or freeze drying.

[0125] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0126] (a) In the solid-state fermentation (SSF) step;

[0127] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0128] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0129] a first incubation thereby obtaining koji, followed by;

[0130] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0131] a second incubation, followed by;

[0132] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0133] (i) directly after cooling or

[0134] (ii) in dry powder form after drying

[0135] (b) In the enzymatic treatment step;

[0136] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;P85424PC01

[0137] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0138] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0139] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0140] (iii) directly after cooling or

[0141] (iv) in dry powder form after drying;

[0142] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0143] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0144] drying to obtain a sweet-umami plant protein product.

[0145] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0146] (a) In the solid-state fermentation (SSF) step;

[0147] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0148] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0149] a first incubation thereby obtaining koji, followed by;

[0150] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0151] a second incubation, followed by;

[0152] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0153] (i) directly after cooling or

[0154] (ii) in dry powder form after drying

[0155] (b) In the enzymatic treatment step;

[0156] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;P85424PC01

[0157] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0158] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0159] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0160] (iii) directly after cooling or

[0161] (iv) in dry powder form after drying;

[0162] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0163] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0164] drying to obtain a sweet-umami plant protein product, wherein step (c) is followed by a 2ndsubmerged fermentation (SMF) step (d) comprising;

[0165] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0166] drying to obtain a sweet-umami plant protein product.

[0167] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0168] (a) In the solid-state fermentation (SSF) step;

[0169] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0170] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0171] a first incubation thereby obtaining koji, followed by;

[0172] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0173] a second incubation, followed by;

[0174] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0175] (i) directly after cooling orP85424PC01

[0176] (ii) in dry powder form after drying

[0177] (b) In the enzymatic treatment step;

[0178] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0179] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0180] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0181] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0182] (iii) directly after cooling or

[0183] (iv) in dry powder form after drying;

[0184] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0185] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0186] drying to obtain a sweet-umami plant protein product, wherein the soaking in the SSF step (a) of the grain and / or legume substrate rich in protein and sweet amino acids is carried out for 0.5-10 h at 4-30 °C.

[0187] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0188] (a) In the solid-state fermentation (SSF) step;

[0189] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0190] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0191] a first incubation thereby obtaining koji, followed by;

[0192] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0193] a second incubation, followed by;

[0194] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:P85424PC01

[0195] (i) directly after cooling or

[0196] (ii) in dry powder form after drying

[0197] (b) In the enzymatic treatment step;

[0198] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0199] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0200] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0201] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0202] (iii) directly after cooling or

[0203] (iv) in dry powder form after drying;

[0204] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0205] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0206] drying to obtain a sweet-umami plant protein product, wherein the concentration of protein of the grain and / or legume substrate is above 20% (w / w), such as between 20-45% (w / w), and the source of sweet amino acids in the SSF step (a) is selected from maize, rice, sorghum, rye, wheat, barley, pearl millet, faba, wheat bran, BSG, and rapeseed / seedpress cake.

[0207] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0208] (a) In the solid-state fermentation (SSF) step;

[0209] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0210] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0211] a first incubation thereby obtaining koji, followed by;

[0212] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;P85424PC01

[0213] a second incubation, followed by;

[0214] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0215] (i) directly after cooling or

[0216] (ii) in dry powder form after drying

[0217] (b) In the enzymatic treatment step;

[0218] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0219] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0220] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0221] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0222] (iii) directly after cooling or

[0223] (iv) in dry powder form after drying;

[0224] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0225] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0226] drying to obtain a sweet-umami plant protein product, wherein the heating in the SSF step (a) of the grain and / or legume substrate is carried out at 60-170 °C for 0.1-3 h followed by cooling to 10-35 °C and thereafter inoculated with 0.01 to 3 g / kg Aspergillus ssp. selected from Aspergillus oryzae, Aspergillus sojae or a combination of Aspergillus oryzae and Aspergillus sojae.

[0227] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0228] (a) In the solid-state fermentation (SSF) step;

[0229] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0230] cooling and subsequent inoculation with Aspergillus spp., followed by;P85424PC01

[0231] a first incubation thereby obtaining koji, followed by;

[0232] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0233] a second incubation, followed by;

[0234] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0235] (i) directly after cooling or

[0236] (ii) in dry powder form after drying

[0237] (b) In the enzymatic treatment step;

[0238] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0239] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0240] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0241] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0242] (iii) directly after cooling or

[0243] (iv) in dry powder form after drying;

[0244] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0245] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0246] drying to obtain a sweet-umami plant protein product, wherein the first incubation in the SST step (a) is carried out at 25-45 °C for 0.5-48 h under 40-100 % of humidity thereby producing koji, followed by the mixing of said koji, grain or legume and amino acids and water in the following ratios 0.1:1:0.1 to 1:3:1, followed by a second incubation carried out at 10-80 °C for 0.5-18 h.

[0247] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0248] (a) In the solid-state fermentation (SSF) step;P85424PC01

[0249] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0250] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0251] a first incubation thereby obtaining koji, followed by;

[0252] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0253] a second incubation, followed by;

[0254] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0255] (i) directly after cooling or

[0256] (ii) in dry powder form after drying

[0257] (b) In the enzymatic treatment step;

[0258] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0259] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0260] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0261] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0262] (iii) directly after cooling or

[0263] (iv) in dry powder form after drying;

[0264] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0265] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0266] drying to obtain a sweet-umami plant protein product, wherein, in the enzymatic treatment step (b), the protein derived from protein-rich grains and / or legumes is protein selected from faba protein, soy protein and oat protein or any combinations thereof added in the mixing step in an amount of 1-20 % (w / w) and the substrate with high fibre concentration, such as 20-40% (w / w), is oat bran added in the mixing step in an amount of 0.1-5 % (w / w), followed by adding water to the mixture an amount of 10-80 % (w / w).P85424PC01

[0267] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0268] (a) In the solid-state fermentation (SSF) step;

[0269] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0270] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0271] a first incubation thereby obtaining koji, followed by;

[0272] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0273] a second incubation, followed by;

[0274] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0275] (i) directly after cooling or

[0276] (ii) in dry powder form after drying

[0277] (b) In the enzymatic treatment step;

[0278] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0279] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0280] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0281] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0282] (iii) directly after cooling or

[0283] (iv) in dry powder form after drying;

[0284] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0285] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0286] drying to obtain a sweet-umami plant protein product, wherein the one or more of proteases, peptidases and glutaminases or combinations thereof added to theP85424PC01

[0287] mixture in the enzymatic treatment step (b) are added to the mixture in amounts of 0.1-4 g / kg at pH 2-9, at 5-80 °C.

[0288] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0289] (a) In the solid-state fermentation (SSF) step;

[0290] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0291] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0292] a first incubation thereby obtaining koji, followed by;

[0293] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0294] a second incubation, followed by;

[0295] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0296] (i) directly after cooling or

[0297] (ii) in dry powder form after drying

[0298] (b) In the enzymatic treatment step;

[0299] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0300] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0301] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0302] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0303] (iii) directly after cooling or

[0304] (iv) in dry powder form after drying;

[0305] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0306] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;P85424PC01

[0307] drying to obtain a sweet-umami plant protein product, wherein the mixing times of the one or more of proteases, peptidases and glutaminases or combinations thereof added to the mixture in the enzymatic treatment step (b) are 2 h for proteases, 1 h for peptidases and 2 h for glutaminases followed by heat treatment of the resulting mixture at 90-150 °C for 1-120 min.

[0308] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0309] (a) In the solid-state fermentation (SSF) step;

[0310] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0311] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0312] a first incubation thereby obtaining koji, followed by;

[0313] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0314] a second incubation, followed by;

[0315] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0316] (i) directly after cooling or

[0317] (ii) in dry powder form after drying

[0318] (b) In the enzymatic treatment step;

[0319] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0320] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0321] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0322] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0323] (iii) directly after cooling or

[0324] (iv) in dry powder form after drying;

[0325] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;P85424PC01

[0326] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0327] drying to obtain a sweet-umami plant protein product, wherein the mixing in the submerged fermentation (SMF) step (c) is carried out as follows: mixing 0.1-50 % (w / w) of the enzymatically treated protein product obtained in the enzymatic treatment step (b) with 0.1-50 % (w / w) of the solid fermented grains / legumes obtained in the SSF step (a).

[0328] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0329] (a) In the solid-state fermentation (SSF) step;

[0330] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0331] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0332] a first incubation thereby obtaining koji, followed by;

[0333] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0334] a second incubation, followed by;

[0335] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0336] (i) directly after cooling or

[0337] (ii) in dry powder form after drying

[0338] (b) In the enzymatic treatment step;

[0339] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0340] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0341] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0342] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0343] (iii) directly after cooling orP85424PC01

[0344] (iv) in dry powder form after drying;

[0345] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0346] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0347] drying to obtain a sweet-umami plant protein product, wherein the one or more alcoholic or non-alcoholic yeasts in the submerged fermentation (SMF) step (c) are Saccharomycodes spp, Saccharomyces spp, Pichia spp., Zygosaccharomyces spp. or any combinations thereof in amounts of 0.1-10 % (w / w).

[0348] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0349] (a) In the solid-state fermentation (SSF) step;

[0350] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0351] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0352] a first incubation thereby obtaining koji, followed by;

[0353] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0354] a second incubation, followed by;

[0355] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0356] (i) directly after cooling or

[0357] (ii) in dry powder form after drying

[0358] (b) In the enzymatic treatment step;

[0359] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0360] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;

[0361] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;P85424PC01

[0362] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0363] (iii) directly after cooling or

[0364] (iv) in dry powder form after drying;

[0365] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0366] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0367] drying to obtain a sweet-umami plant protein product, wherein the fermentation under non-static and anaerobic conditions in SMF step (c) is carried out for 2-96 h, at 1-55 °C, followed by pasteurization carried at 60-140 °C, for 0.01-60 min.

[0368] Still another embodiment of the invention relates to a process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:

[0369] (a) In the solid-state fermentation (SSF) step;

[0370] heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;

[0371] cooling and subsequent inoculation with Aspergillus spp., followed by;

[0372] a first incubation thereby obtaining koji, followed by;

[0373] mixing said koji with grain or legume rich in protein and amino acids and water, followed by;

[0374] a second incubation, followed by;

[0375] the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:

[0376] (i) directly after cooling or

[0377] (ii) in dry powder form after drying

[0378] (b) In the enzymatic treatment step;

[0379] mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;

[0380] adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;P85424PC01

[0381] inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;

[0382] the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:

[0383] (iii) directly after cooling or

[0384] (iv) in dry powder form after drying;

[0385] (c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;

[0386] inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;

[0387] drying to obtain a sweet-umami plant protein product, wherein the following additional steps are added to the SMF step (c):

[0388] Inoculation with 0.1-10 % (w / w) of one or more of Torula spp., Candida magnoliae, Yarrowia lipolytica, Propionibacterium spp. and lactic acid bacteria, such as Streptococcus spp., Lactococcus spp., Lactobacillus spp., followed by;

[0389] SMF fermentation for 2-48 h at 1-50 °C under static or non-static and anaerobic or aerobic conditions.

[0390] Still another embodiment of the invention relates to a food product comprising a fermented sweet-umami plant-protein ingredient containing at least 30% protein and providing sweet and umami taste together with fruity, honey, caramel, nutty, caramel and / or roasted aromas and further comprising one or more polyalcohols selected from erythritol, mannitol, and glycerol, wherein the food product (i) shows same or better flavor characteristics compared to a control product without the sweet-umami plant-protein ingredient and / or (ii) provides increased creaminess at an equivalent sweet-taste level without requiring higher sugar content and / or (iii) masks the off / bitter taste of plant-protein.

[0391] In still another embodiment, the drum drying, spray drying or freeze drying steps of the present invention is carried out by subjecting the product to heat treatment at a temperature at 90-100°C for 10-40 minutes followed by a cool down to < 8°C and directly transferred to the downstream processing, which e.g. could be centrifugation, spray drying, hotfill, fluidized-bed dryer in order to provide a final product material in either liquid, paste or powder form.P85424PC01

[0392] In still another embodiment relates to sweet umami plant-protein product produced by the process of the invention.

[0393] In still another embodiment relates to a food product comprising a sweet umami plant-protein product produced by the process of the invention having honey, caramel, nutty, and roasted flavors with the same sweet taste level compared to a food product with higher sugar content and / or having increased creaminess compared to a control food product without sweet umami plant protein.

[0394] In still another embodiment relates to a sweet tasting plant-protein product produced by fermentation, wherein the plant-protein product has umami and volatile aroma compounds such as honey and / or caramel (acetate esters, including hexyl acetate, butyl acetate, and 4-pentenyl acetate, or aldehydes such as hexanal, nonanal or benzaldehyde) and the plant-protein product comprises at least 30% protein.

[0395] Still another aspect of the present invention relates to a food product comprising a sweet umami plant-protein product produced by the process of the invention having honey, fruity, caramel, nutty, and roasted aromas (acetate esters, including hexyl acetate, butyl acetate, and 4-pentenyl acetate, or aldehydes such as hexanal, nonanal or benzaldehyde ), compared to a control food product with higher sugar content, without compromising sweet taste.

[0396] Still another aspect of the present invention is to provide a food product comprising the sweet umami plant-protein product produced by the process of the invention, having increased creaminess compared to a control food product with higher sugar content, without compromising sweet taste.

[0397] It should be noted that embodiments and features described in the context of one of the aspects of the present invention also apply to the other aspects of the invention.

[0398] All patent and non-patent references cited in the present application, are hereby incorporated by reference in their entirety.P85424PC01

[0399] The invention will now be described in further details in the following non-limiting examples.

[0400] Examples

[0401]

[0402] Enzymatic treatment

[0403] 7 % (w / w) faba protein was added along with 63 % (w / w) of water and 3 % (w / w) oat bran, and all ingredients were mixed thoroughly in a mixer. The enzyme mixture was then added, and the pH was adjusted to 4.5. The reaction was conducted at 37 °C for 5 h, with specific mixing times for the enzymes: 2 h for proteases, 1 h for peptidase, and 2 h for glutaminase. The enzyme concentration for each enzyme was 0.2 g / kg. The proteases included were Proteases P "Amano" 6SD in a concentration of 0.2 g / kg, Proteases HF "Amano" 150SD in a concentration of 0.2 g / kg, and ProteAX (EC 3.4.21). The peptidase used was Peptidase R in a concentration of 0.2 g / kg (EC 3.2.1), and glutaminase (EC 3.5.1) in a concentration of 0.2 g / kg was also added. The enzymes were inactivated through heat treatment at 95 °C for 15 minutes. After enzyme inactivation, the final product was used either liquid or solid after drying process. Freeze-drying or spray-drying was used to obtain a stable powder.

[0404] Solid-state fermentation (SSF)

[0405] The SSF step involves soaking of 1 kg of oats for 4 h at a temperature range of 22 °C, followed by rinsing with cold water. The soaked oats were then cooked in an oven at 100 °C for 20 min, after which they were cooled to a temperature of 32 °C. Once cooled, the oats were inoculated with Aspergillus sojae and Aspergillus oryzae in a concentration of 0.3 g / kg and incubated at 32 °C for 40 h under a relative humidity of 60 %. The inoculated oat substrate was mixed with koji and water in a ratio of 1:2:1 (oat: koji: water). The mixture was fermented in the incubator at 55 °C for 8 h. Upon completion of fermentation, the resulting product was either freeze-dried or spray-dried to obtain a powdered form.

[0406] 1ststage Submerged fermentation (SMF)P85424PC01

[0407] The intermediate product obtained in the enzymatic treatment step (7 % (w / w)) was mixed with solid fermented grains / legumes (28.5 % (w / w)) obtained in the SSF step. The mixture was inoculated with one or more species from Saccharomyces pastorianus, Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans, at a concentration of 1.5 % (w / w). Fermentation was conducted for 16 h under non-static and anaerobic conditions. After fermentation, the product was pasteurized at 100 °C for 21 minutes, followed by either freeze- drying or spray-drying to obtain a final powder form.

[0408] Optional: 2ndstage Submerged fermentation (SMF)

[0409] The mixture was inoculated with one or more species from Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans, s at a concentration of 1.5 % (w / w). Fermentation was conducted for 4 h under non-static and anaerobic conditions. After fermentation, the product was pasteurized at 100 °C for 21 minutes, followed by either freeze-drying or spray-drying to obtain a final powder form.

[0410]

[0411] Enzymatic treatment

[0412] 7 % (w / w) faba protein was added along with 63 % (w / w) of water and 3 % (w / w) of oat bran, and all ingredients were mixed thoroughly in a mixer. The enzyme mixture was then added, and the pH was adjusted to 4.5. The reaction was conducted at 37 °C for 5 h, with specific mixing times for the enzymes: 2 h for proteases, 1 h for peptidase, and 2 h for glutaminase. The enzyme concentration was 0.2 g / kg. The proteases included were Proteases P "Amano" 6SD to 0.2 g / kg, Proteases HF "Amano" 150SD to 0.2 g / kg, and ProteAX (EC 3.4.21) to 0.2 g / kg. The peptidase used was Peptidase R to 0.2 g / kg (EC 3.2.1), and glutaminase to 0.2 g / kg (EC 3.5.1) was also added. The enzymes were inactivated through heat treatment at 95 °C for 15 min. After enzyme inactivation, the final product was subjected to either a liquid process or a drying process.

[0413] Solid-state fermentation (SSF)P85424PC01

[0414] The SSF step involves soaking of oats for 4 h at a temperature range of 22 °C, followed by rinsing with cold water. The soaked oats were then cooked in an oven at 100 °C for 20 min, after which they were cooled to 31 °C. Once cooled, the oats were inoculated with Aspergillus oryzae to 0.3 g / kg and incubated at 32 °C for 40 h under a relative humidity of 60 %. The inoculated oat substrate was mixed with koji and water in a ratio of 1:2:1 (oat : koji: water). The mixture was fermented in the incubator at 55 °C for 8 h. Upon completion of fermentation, the resulting product was either freeze-dried or spray-dried to obtain a powdered form.

[0415] 1ststage submerged fermentation (SMF): produce B12, and sugar alcohol.

[0416] The intermediate product obtained in the enzymatic treatment step was mixed with 28.5 % (w / w) solid fermented oat. The mixture was then inoculated with Propionibacterium spp. and Lactobacillus ssp. at a concentration of 0.8 % (w / w). The fermentation process was carried out for 16 h under static and aerobic conditions.

[0417] 2ndstage submerged fermentation (SMF): synergistically enhancing aroma compounds and -profile

[0418] The mixture resulting from 1ststage submerged fermentation was inoculated with one or more species from Saccharomyces pastorianus, Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans, at a concentration of 1.5 % (w / w). Fermentation was conducted for 16 h under static and anaerobic conditions. After fermentation, the product was pasteurized at 100 °C for 21 min, followed by either freeze-drying or spray-drying to obtain a final powder form.

[0419] Optional: 3rdstage Submerged fermentation (SMF)

[0420] The mixture was inoculated with one or more species from Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans, at a concentration of 1.5 % (w / w). Fermentation was conducted for 4 h under non-static and anaerobic conditions. After fermentation, the product was pasteurized at 100 °C for 21 minutes, followed by either freeze-drying or spray-drying to obtain a final powder form.

[0421] Example 3 - Applications / usesP85424PC01

[0422] The sweet umami plant-protein product produced according to the process of the invention can be used for multiple purposes, such as, but not limited to:

[0423] Bakery category: includes products such as bars, breads, buns, cookies, cakes, crackers.

[0424] - Sport / nutrition goods: smoothie, bars.

[0425] Ready to eat sweet type products

[0426] Dairy alternatives such as milk, yogurt or ice creams

[0427] Plant-based products

[0428] Example 4 - Product analysis and results

[0429] The sweet umami plant-protein product produced according to the process of the invention contains the following sugars in the following amounts:

[0430] Sugar content:

[0431] - Sugars: 3.54 g / lOOg

[0432] - Fructose 0.4 g / lOOg

[0433] - Galactose < 0.1 (LOQ)g / 100g

[0434] - Glucose 0.5 g / lOOg

[0435] - Lactose 2.0 g / 100g

[0436] - Maltose 0.3 g / 100g

[0437] - Sucrose 0.3 g / 100g

[0438] The sweet umami plant-protein product produced according to the process of the invention contains the following proteins in the following amounts:

[0439] Protein content

[0440] • HP12 NMKL 6:2003 mod. I Kjeldahl (titrimetry): Protein (N*6,25) 44.2 g / 100 g

[0441] • Volatic organic compounds: at a concentration of 1.5 % (w / w).

[0442] Fermentation was conducted for 4 h under non-static and anaerobic conditions. After fermentation, the product was pasteurized at 100 °C for 21 minutes, followed by either freeze-drying or spray-drying to obtain a final powder form.

[0443] A relatively large number of aroma compounds were found to contribute to the enhancement of sweet or sweet-umami taste perception. Among the identified alcohols, 1-propanol was detected, with its concentration varyingP85424PC01

[0444] depending on the yeast strain used, indicating strain-dependent differences in aroma production.

[0445] Esters also played a key role in modulating sweetness perception. Several acetate esters, including hexyl acetate, butyl acetate, and 4-pentenyl acetate, were identified, showing distinct concentration patterns across the different yeast fermentations. Ethyl esters further contributed to sweet aroma notes, with ethyl acetate being widely present, while ethyl hexanoate and ethyl octanoate appeared at varying levels depending on the yeast employed.

[0446] Two fatty acids— isobutyric acid and isovaleric acid— were detected in this analysis. In addition, several aldehydes with fruity or nutty odor characteristics were identified. Hexanal, associated with green and fruity notes, benzaldehyde, which imparts almond-like aromas, and nonanal, known for citrus peel notes, were all detected at different concentrations across the yeast variants. Despite these quantitative differences, all these compounds collectively contribute to the enhancement of sweet perception in the fermented products (Figure 3 and Table 1).

[0447]

[0448] P85424PC01

[0449]

[0450] P85424PC01

[0451]

[0452] Table 1. Volatile aroma compounds from fermentation with four different yeast Saccharomyces pastorianus, Pichia kluyveri, Torulaspora delbrueckii, Lanchancea thermotolerans) and the description of specific compounds. Fermentation was performed at 25 °C, anaerobic conditions and non-static for 16 h and analysis by Dynamic Headspace Gas chromatography / Mass Spectrometry (DHS / GC-MS).

[0453] • Polyalcohols: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high-resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Erythritol, maltitol, threitol, d-arabitol and glycerol were identified and increased after the fermentation compared to the sample without fermentation, only the mixture of all raw ingredients and microorganisms (Figure 4).

[0454] • Sweet amino acids: The amino acid panel was analysed using a GC-MS method in which the target amino acids were derivatized with methyl chloroformate (MCF) to generate volatile compounds suitable for GC-MS detection. Glycine, alanine, serine, glutamine, and threonine and tryptophan were identified and increased after the fermentation process (Figure 5).P85424PC01

[0455] • Umami amino acids: The amino acid panel was analysed using a GC-MS method in which the target amino acids were derivatized with methyl chloroformate (MCF) to generate volatile compounds suitable for GC-MS detection. Glutamic acid, the main contributor to umami taste, was identified, and its concentration increased two time after fermentation (Figure 6).

[0456] • Kokumi peptides and bitter amino acids: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high-resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Kokumi peptides formatted with bitter amino acids, such as valine (Vai), phenylalanine (Phe), leucine (Leu), increased during the fermentation, showing a way to decrease the bitter taste of amino acids in the sweet umami plant protein through the transpeptidation reaction during fermentation (Figure 7).

[0457] • Kokumi peptides identification: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high-resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Different -glutamyl peptides were identified, such as y-Glu-Phe, y-Glu-Glu, y-Glu-Leu, y-Glu- Ile, y-Glu-Val, y-Glu-Val-Gly and y-Glu-Gly (Figure 8).

[0458] • Vitamins: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high- resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Vitamin Bl (Thiamine), vitamin B2 (Riboflavin), vitamin B5 (Pantothenic acid), and vitamin B7 (Biotin) were identified and increased after the fermentation (Figure 9).

[0459] • Postbiotic compounds: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high-resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Nine different compounds were identified as a postbiotic compounds, such as lactic acid, succinic acid, citric acid, tyramine, y-aminobutyric acid (GABA), agmatine, phenylacetic acid, kojic acid and caffeic acid, and increased after fermentation (Figure 10A).P85424PC01

[0460] Additionally, 2-methylpropanoic acid, (short chain fatty acid) was identified and increased after fermentation (Figure 10B).

[0461] • Prebiotic compounds: The analysis was performed in randomized sample order using a UHPLC system (Vanquish, Thermo Fisher Scientific) coupled to a high-resolution quadrupole-Orbitrap mass spectrometer (Orbitrap Exploris 240, Thermo Fisher Scientific). Caffeic acid, p-coumaric acid, syringin, D-raffinose, satchyose and nystose where identified in the samples, and they increased after fermentation (Figure 11).

[0462] The sweet umami plant-protein product produced according to the process of the invention comprises the following functional properties.

[0463] In protein bar formulations, the ingredient demonstrated high water-holding capacity, enabling effective moisture retention and contributing to structural stability without free water separation. This functionality supported cohesive binding of bar components and reduced brittleness during handling and storage. The ingredient also exhibited good solubility upon hydration, comparable to isolate protein powders, facilitating uniform dispersion during mixing and contributing to a homogeneous final matrix.

[0464] Texturally, the ingredient promoted the formation of a smooth and continuous protein network, yielding a silky, paste-like consistency. The internal structure of the bar was homogeneous and free from grittiness, and during consumption the texture transitioned into a creamy, smoothie-like mouthfeel. These properties indicate favorable hydration behavior and protein-matrix interactions, allowing the ingredient to function simultaneously as a protein source, moisture regulator, and texture modifier in high-protein bar applications.

[0465] In biscuit formulations, the ingredient was observed to accelerate Maillard reactions during baking, as evidenced by earlier onset of browning and enhanced surface color development under comparable thermal conditions. This behavior suggests increased availability of reactive amino compounds within the protein matrix, contributing to intensified thermal reactivity. The resulting biscuits exhibited a pronounced and desirable crispiness, with a clean fracture and dry, brittle texture upon biting. Overall, the ingredient supported uniform baking,P85424PC01

[0466] appealing color formation, and improved textural quality, highlighting its multifunctional role in baked applications.

[0467] Example 5 - Comparison product tests relating to sensory and functionality Color (Maillar reaction)

[0468] Butter cookies were used as a model for color comparison. The cookies were made with sugar, flour, egg, and butter, mixed together, and baked for 15 minutes at 180 °C. In samples containing the sweet umami plant-protein product obtained according to the process of the invention, the Maillard reaction is noticeably higher and faster than in the control (Figure 12). The sweet-umami plant-protein product accelerates the Maillard reaction, which can decrease baking time across different processes while preserving the desired aroma, texture, flavor, and overall sensory profile (refer to the consumer sensory analysis below).

[0469] Example 6 - Consumer sensory analysis in biscuits

[0470] Butter cookies (made with sugar, flour, egg, and butter) were used as a model for consumer sensory analysis. The sensory analysis was divided into two parts. The first part utilized the CATA (Check-AII-That-Apply) method to study the flavor profile of each sample, with 21 attributes used to describe all samples, as shown in Figure 13.

[0471] The butter cookies were made using sugar (12.7 % (w / w)), flour (50.6 % (w / w)), and egg white (25.3 % (w / w)) as the control recipe (171). For samples 357 and 513, egg white was replaced with 25.3 % (w / w) water, and sugar was partially replaced 1:1 by sweet umami plant-protein prepared according to the process of the present invention, reducing sugar content by 35 % (w / w) and 20 % (w / w), respectively, using the process without B12 production (Figure 1). For samples 715 and 289, the same recipe was followed, with sugar reduced by 35 % (w / w) and 20 % (w / w) and replaced 1:1 with sweet umami plant-protein prepared according to the process of the present invention (Table 2).

[0472] In the second part of this experiment the intensity of sweetness, umami, bitter aftertaste, and creaminess was evaluated. Additionally, the sense of liking was assessed in terms of aroma, texture, and taste, with specific attributes identified to describe each sample. The following samples were used.P85424PC01

[0473]

[0474] Table 2. Samples used in the sensory analysis were prepared according to the process described above, with reduced sugar and egg replaced as outlined in the description.

[0475] In this study, sugar was reduced by 20 % and 35 %. Figure 14 shows that samples 171, 513, and 715 are located in the same region of the plot. Samples 171 and 513 are particularly close to each other, sharing attributes such as honey, caramel, nutty, and roasted flavors, along with a brown color. In contrast, samples 289 and 357, which have a 35 % sugar reduction, exhibit flavors that are significantly different from the control sample (171). For this reason, only samples 171, 715, and 513 were selected for the remainder of the study.

[0476] According to the statistical analysis, samples 171 and 513 do not show any significant differences in the intensity of sweetness, bitter aftertaste, and umami; however, they do show a significant difference in creaminess (Table 3). The creaminess of sample 513 was perceived as higher than that of the control sample (171). When comparing samples 715 and 171, Table 3 (below) indicates differences in sweetness, umami, and creaminess intensities, with sample 715 being higher in umami and creaminess, while sweetness is higher in the control sample (171) (Figure 15).P85424PC01

[0477] >

[0478]

[0479] intensity perception in a 1-5 scale for sweet, umami, creaminess and bitter aftertaste. Note: bold font indicates significant p-values (p < 0.05).

[0480] In term of liking, samples 513 and 171 (control) do not show any significant differences for overall of taste, aroma and texture, however sample 715 shows differences with the control samples (table 4) (Figure 16 and Figure 18).

[0481]

[0482] Table 4. Results of two-way ANOVA and post hoc analysis (Tukey HSD) of liking in a 1-9 liking-scale for aroma, taste and aroma. Note: bold font indicates significant p-values (p < 0.05).

[0483] The consumer sensory analysis shows that the sweet umami plant-protein product can reduce sugar content by 20 % (w / w) and decrease fat concentration by enhancing the perception of creaminess. It also improves umami taste and introduces flavors such as caramel, honey, and roasted notes, similar to the control sample (Figure 16 and Figure 18).

[0484] Example 9 - tests on the texture of the final product

[0485] The data in this example is from the consumer sensory analysis. Sample 171 (control) was rated the highest for crispiness, followed by a crumbly texture. TheP85424PC01

[0486] remaining samples containing the sweet umami plant-protein product were rated as having a softer texture. In the liking test, the texture of Sample 171 (control) and Sample 513 (with a 20 % sugar reduction) were rated similarly, showing no significant differences between them (Figure 17).

[0487] Example 10 - Consumer sensory analysis in protein bar

[0488] A protein bar was prepared by first mixing the dry ingredients (oat protein, pea protein, whole oat, salt, and lecithin). Separately, a binder phase of brown rice syrup, water, sunflower oil, peanut butter, and date paste was gently heated and A protein bar was prepared by first mixing the dry ingredients (oat protein, pea protein, whole oat, salt, and lecithin). Separately, a binder phase of brown rice syrup, water, sunflower oil, peanut butter, and date paste was gently heated and combined until homogeneous. The binder was added to the dry blend and mixed to form a cohesive mass, which was then pressed into molds and allowed to set. A control formulation was produced without the sweet-umami plant protein. In the experimental formulations, different yeast-derived sweet-umami proteins were incorporated by replacing a portion of the plant protein blend at the dry-mixing stage. All samples were processed identically to enable comparison of texture and flavor attributes.

[0489] Protein bar formulations for different samples

[0490] Protein bar samples were evaluated by 30 participants. Most of the participants were female between 18 to 29 years old and omnivores. The most important parameter in consumers' choice seems to be the taste and protein content. They also consider healthy ingredients among others. Interestingly it seems the sugar content is not a concern for the consumers of protein bars as much as other parameters. The rank rating results showed that SUPP 72 and 71 had higher umami perception compared to reference. The highest sweet taste was perceived at SUPP 71 (Figure 19). These results led to the highest liking score of SUPP 71 protein bar. It is worth noting that SUPP 71 showed slightly lower bitter after taste but not significantly different from other prototypes (Figure 20).

[0491]

[0492] P85424PC01

[0493]

[0494] Table 5. Samples used in the sensory analysis were prepared according to the process described above, with reduced sugar and egg replaced as outlined in the description.P85424PC01

[0495] References

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Claims

55P85424PC01Claims1. Process for producing sweet umami plant-protein product, wherein the process comprises at least one solid-state fermentation (SSF) step, an enzymatic treatment step and a submerged fermentation (SMF) step, wherein said steps are carried out as follows:(a) In the solid-state fermentation (SSF) step;heating of a soaked and rinsed grain and / or legume substrate rich in protein and sweet amino acids, followed by;cooling and subsequent inoculation with Aspergillus spp., followed by; a first incubation thereby obtaining koji, followed by;mixing said koji with grain or legume rich in protein and amino acids and water, followed by;a second incubation, followed by;the resulting product from step (a) is applied into the submerged fermentation (SMF) step (c) either:(i) directly after cooling or(ii) in dry powder form after drying(b) In the enzymatic treatment step;mixing of protein derived from protein-rich grains and / or legumes with a substrate rich in fibre and water, followed by;adding one or more of proteases, peptidases and glutaminases or combinations thereof to the mixture, followed by;inactivation of said enzymes from the enzymatically treated mixture by heat treatment, followed by;the resulting product from step (b) is applied into the submerged fermentation (SMF) step (c) either:(iii) directly after cooling or(iv) in dry powder form after drying;(c) In the submerged fermentation (SMF) step; mixing the products obtained in (a) and (b), followed by;inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;56P85424PC01drying to obtain a sweet-umami plant protein product.

2. The process according to claim 1, wherein step (c) is followed by a 2ndsubmerged fermentation (SMF) step (d) comprising;inoculation with one or more alcoholic or non-alcoholic yeasts, followed by; fermentation under non-static and anaerobic conditions, followed by; pasteurization, followed by;drying to obtain a sweet-umami plant protein product.

3. The process according to any of claims 1-2, wherein the soaking in the SSF step (a) of the grain and / or legume substrate rich in protein and sweet amino acids is carried out for 0.5-10 h at 4-30 °C.

4. The process according to any of claims 1-3, wherein the concentration of protein of the grain and / or legume substrate is above 20% (w / w), such as between 20-45% (w / w), and the source of sweet amino acids in the SSF step (a) is selected from maize, rice, sorghum, rye, wheat, barley, pearl millet, faba, wheat bran, BSG, and rapeseed / seedpress cake.

5. The process according to any of claims 1-4, wherein the heating in the SSF step (a) of the grain and / or legume substrate is carried out at 60-170 °C for 0.1-3 h followed by cooling to 10-35 °C and thereafter inoculated with 0.01 to 3 g / kg Aspergillus ssp. selected from Aspergillus oryzae, Aspergillus sojae or a combination of Aspergillus oryzae and Aspergillus sojae.

6. The process according to any of claims 1-5, wherein the first incubation in the SST step (a) is carried out at 25-45 °C for 0.5-48 h under 40-100 % of humidity thereby producing koji, followed by the mixing of said koji, grain or legume and amino acids and water in the following ratios 0.1:1:0.1 to 1:3:1, followed by a second incubation carried out at 10-80 °C for 0.5-18 h.

7. The process according to any of claims 1-6, wherein, in the enzymatic treatment step (b), the protein derived from protein-rich grains and / or legumes is protein selected from faba protein, soy protein and oat protein or any combinations thereof added in the mixing step in an amount of 1-20 % (w / w) and57P85424PC01the substrate with high fibre concentration, such as 20-40% (w / w), is oat bran added in the mixing step in an amount of 0.1-5 % (w / w), followed by adding water to the mixture an amount of 10-80 % (w / w).

8. The process according to any of claims 1-7, wherein the one or more of proteases, peptidases and glutaminases or combinations thereof added to the mixture in the enzymatic treatment step (b) are added to the mixture in amounts of 0.1-4 g / kg at pH 2-9, at 5-80 °C.

9. The process according to any of claims 1-8, wherein the mixing times of the one or more of proteases, peptidases and glutaminases or combinations thereof added to the mixture in the enzymatic treatment step (b) are 2 h for proteases, 1 h for peptidases and 2 h for glutaminases followed by heat treatment of the resulting mixture at 90-150 °C for 1-120 min.

10. The process according to any of claims 1-9, wherein the mixing in the submerged fermentation (SMF) step (c) is carried out as follows: mixing 0.1-50 % (w / w) of the enzymatically treated protein product obtained in the enzymatic treatment step (b) with 0.1-50 % (w / w) of the solid fermented grains / legumes obtained in the SSF step (a).

11. The process according to any of claims 1-10, wherein the one or more alcoholic or non-alcoholic yeasts in the submerged fermentation (SMF) step (c) are Saccharomycodes spp, Saccharomyces spp, Pichia spp., Zygosaccharomyces sp p. or any combinations thereof in amounts of 0.1-10 % (w / w).

12. The process according to any of claims 1-11, wherein the fermentation under non-static and anaerobic conditions in SMF step (c) is carried out for 2-96 h, at 1-55 °C, followed by pasteurization carried at 60-140 °C, for 0.01-60 min.

13. The process according to any of claims 1-12, wherein the following additional steps are added to the SMF step (c):Inoculation with 0.1-10 % (w / w) of one or more of Torula spp., Candida magnoliae, Yarrowia lipolytica, Propionibacterium spp. and lactic acid bacteria, such as Streptococcus spp., Lactococcus spp., Lactobacillus spp., followed by;P85424PC01SMF fermentation for 2-48 h at 1-50 °C under static or non-static and anaerobic or aerobic conditions.

14. A food product comprising a fermented sweet-umami plant-protein ingredient containing at least 30% protein and providing sweet and umami taste together with fruity, honey, caramel, nutty, and / or roasted notes and further comprising one or more polyalcohols selected from erythritol, mannitol, and glycerol, wherein the food product (i) shows some or better flavor characteristics compared to a control product without the sweet-umami plant-protein ingredient and / or (ii) provides increased creaminess at an equivalent sweet-taste level without requiring higher sugar content and / or (iii) masks the off / bitter taste of plantprotein.

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