Plant-based food products and process for obtaining thereof

Abstract

The present invention refers to a process for obtaining plant-based food products, preferably plant-based meat, without plant flavour off-notes, or with reduced plant flavour off-notes, as compared with the plant of origin. The present invention also refers to plant-based food products, preferably plant-based meat, obtained or obtainable by using or implementing the process of the invention.

Description

[0001] PLANT-BASED FOOD PRODUCTS AND PROCESS FOR OBTAINING THEREOF

[0002] FIELD OF THE INVENTION

[0003] The present invention refers to the biotech field, particularly applied to the food industry. Specifically, the present invention refers to a process for obtaining plant-based food products, preferably plant-based meat, without plant flavour off-notes, or with reduced plant flavour off- notes as compared with the plant of origin. The present invention also refers to plant-based food products, preferably plant-based meat, obtained or obtainable by using or implementing the process of the invention.

[0004] STATE OF THE ART

[0005] The global projections show that feeding a world population of almost 10 billion people in 2050 would require raising overall food production by 70 percent. Currently, this food gap is being related to unsustainable agri-food practices. At the same time, we urgently need to cut greenhouse gas (GHG) emissions from agricultural production and stop conversion of remaining forests to agricultural land. Thus, there is an enormous interest in developing new sources of food, sustainable, nutritional and healthy.

[0006] Use of microorganisms as an alternative source of SCP (Single Cell Proteins) arises with great potential due to all the advantages they present, such as their independence from seasonal or climatic factors and the use of facilities in very small areas, which reduces 25 deforestations caused by other alternatives (vegetable and / or animal). In addition, the water consumed, and the CO2 emitted into the atmosphere are reduced compared to animal protein (approximately 98% and 85% respectively). Much of the recent interest in SCP has focused on the valorization of side streams by using microorganisms to improve their protein content, which can then be used in animal feed and human consumption.

[0007] Meat analogues, need to mimic certain qualities of meat, and are blends of various protein sources. Texturization processes are used to develop meat analogues from various protein sources to replace animal proteins in human. Extrusion technologies are the most common texturization processes. With these technologies the fibre like structure characteristic of meet can be mimic.

[0008] Currently plant-based protein derived meat analogues lead the transition towards a more sustainable food system. While consumers are increasing their awareness of the environmental impact of meat consumption, only a fraction of the population chooses a plant-based diet. The primary factor behind this choice is that existing formulations of plant-based protein-derived alternative meats encounter several challenges, particularly in terms of organoleptic properties. In order to arise the meat consumption, plant-based products should be formulated to mimic meat. The lack of meaty aroma and taste and the off flavours are considered potential constrains for product formulations. A recent survey showed the low consumer acceptance is likely related to the poor sensory properties of plant-based products. Thus, the need of an ingredient that can improve this characteristic is necessary.

[0009] The research to mitigate off-flavors in plant-based proteins is of high importance, particularly considering their increasing utilization in food applications. These proteins often exhibit off- notes such as "green," "grassy," or "beany" flavors, alongside persistent bitter and astringent tastes, which represent significant challenges to consumer acceptance and marketability. In addressing this flavor challenge, it's essential to recognize the inherent limitations of synthetic flavoring agents commonly used in food products Unlike the characteristic flavors of specific types of meat products, synthetic flavorings lack the complexity and richness associated with natural sources. Moreover, synthetic flavorings are short-lived and susceptible to degradation during cooking in extreme conditions, making them incapable of replicating the flavor changes experienced in cooked meats. Importantly, the use of synthetic flavorings may also raise concerns regarding human health, as they have the potential to accumulate in the body over time, leading to the development of toxic compounds such as carcinogens, mutagens, and teratogens.

[0010] Simultaneously, the integration of aroma compounds into food matrices is a growing area of concern. Aroma compounds are often expensive, volatile, and chemically unstable under thermal and mechanical stress, leading to significant losses during industrial processing. This not only diminishes product quality and consumer acceptance but also increases formulation costs due to the need for over-dosing or post-process flavoring, the stabilization and preservation of aroma profiles during harsh processing conditions is a critical challenge, especially in the development of shelf-stable, plant-based food products. Aroma compounds are among the most chemically sensitive components in food formulations, being highly volatile and susceptible to thermal degradation, oxidation, and polymerization under high temperature and pressure conditions. For instance, during pasteurization, sterilization, or extrusion cooking, the exposure to elevated thermal and mechanical stress frequently results in partial or complete loss of top-note volatiles. This degradation affects both product quality and consumer perception, ultimately reducing product differentiation and increasing cost due to aroma overcompensation.

[0011] Current approaches to address these issues include encapsulation technologies, enzymatic treatments, and the addition of masking agents. However, these methods are often limited in scope, not compatible with all processing conditions, or insufficiently robust to survive the harsh environments of thermal processing.

[0012] Therefore, there remains a critical need for functional food ingredients that can simultaneously mask plant-protein-derived off-notes and preserve aromatic integrity through industrially relevant conditions such as extrusion and pasteurization. The present invention addresses this gap by providing a novel ingredient with dual functionality: aroma protection and sensory improvement in plant-based formulations.

[0013] DESCRIPTION OF THE INVENTION

[0014] The present invention refers to a process for obtaining plant-based food products, preferably plant-based meat, without plant flavour off-notes, or with reduced plant flavour off-notes as compared with the plant of origin. The present invention also refers to plant-based food products, preferably to plant-based meat, obtained or obtainable by using or implementing the process of the invention.

[0015] Particularly, the present invention refers to a process which comprise the use of inactivated yeasts to mask undesirable sensory attributes ("off-notes") and / or protecting volatile aroma compounds during industrial food processing. More particularly, the invention pertains to solutions applicable to plant-based protein systems subjected to high-temperature and high- pressure conditions such as pasteurization, sterilization, and extrusion, wherein aroma degradation and the development of off-flavours are prevalent challenges.

[0016] So, the inventors of the present invention herein provide an innovative approach aimed at obtaining plant-based food products, preferably meat, without plant flavour off-notes, or with reduced plant flavour off-notes as compared with the plant of origin.

[0017] Particularly, the present invention provides a novel ingredient (specific inactivated yeasts) composition that simultaneously (i) protects aroma compounds from degradation under thermal and / or mechanical stress and (ii) reduces or masks undesirable sensory attributes ("off-notes") commonly associated with plant-based proteins. The composition is designed to be functionally stable during industrial food processing techniques, including but not limited to pasteurization, sterilization (retort), and extrusion cooking. The invention leverages the synergistic interaction between edible yeast components to form a protective matrix or interface that encapsulates volatile aromatic compounds while interacting with and neutralizing reactive off-flavor precursors present in protein systems.

[0018] This dual-functionality allows for: i) Improved aroma retention throughout processing and storage, ii) enhanced sensory profile of plant-based foods by reducing bitterness, astringency, and beany notes, iii) cost efficiency by minimizing aroma loss and reducing the need for postprocess flavoring and iv) compatibility with existing industrial processes without requiring additional technological investment.

[0019] Consequently, the present invention, apart from providing plant-based food products, preferably plant-based meat, without plant flavour off-notes, or with reduced plant flavour off- notes; it provides plant-based food products, preferably plant-based meat, with improved aroma retention, particularly flavourings that impart a realistic meat flavour to products where real meat may be absent or used in limited amounts (i.e., poultry flavour). In this regard, the present invention provides results with:

[0020] • The "aroma of poultry" (FA1) from Hela (Hela Gewiirzwerk Hermann Laue GmbH): It is a food-grade flavouring specifically formulated to mimic the characteristic taste and smell of cooked poultry, such as chicken or turkey. It is likely composed of a blend of flavouring compounds, including protein hydrolysates that contribute umami notes, and possibly sulphur-containing molecules that replicate the roasted or boiled meat profile typical of poultry. This type of aroma is commonly used in industrial food production to enhance or recreate meaty flavours in ready meals, soups, sauces, savoury snacks, or even in vegetarian or vegan products that aim to simulate the taste of meat.

[0021] • SAVAROME P400 (FA2) from Kerry (Kerry Group pic) is also a complex food flavouring designed to deliver a rich, meaty flavour, probably with a focus on poultry or white meat profiles. Developed by Kerry, a company known for its advanced flavour systems, this aroma likely combines cooked meat notes with subtle roasted or umami tones and may include spice or toasted undertones for depth. It is typically used in processed foods and prepared dishes to improve palatability and give a more authentic meat-like experience, especially when the product formulation is low in natural meat content or completely plant-based. Therefore, the invention is applicable across various product categories, including plant-based meats, dairy alternatives, protein-enriched beverages, soups, sauces, and functional snacks.

[0022] So, the first embodiment of the present invention refers to a process (hereinafter process of the invention) for obtaining plant-based food products, preferably plant-based meat, without plant flavour off-notes, or with reduced plant flavour off-notes as compared with the plant of origin; or a process for mitigating or eliminating plant flavour off-notes in plant-based food products, preferably in meat analogues, which comprises: a) providing a source of plant protein, and b) mixing the plant protein with inactivated yeast biomass.

[0023] In a preferred embodiment, the process of the invention is characterized in that fermentation or extraction steps are not carried out, so the process is easier and more cost-effective as compared with the state of the art.

[0024] In a preferred embodiment, the process of the invention is characterized in that it further comprises subjecting the mixture of step b) to pressure and / or temperature, preferably pressure between 0.5 and 50 bar and / or temperature between 60 and 200 °C.

[0025] In a preferred embodiment, the process of the invention is characterized in that it further comprises extruding a mixture of the plant protein with yeast biomass.

[0026] In a preferred embodiment, the process of the invention is characterized in that plant-based food products characterized by the retention of added meaty flavour compounds are obtained.

[0027] Unlike traditional methods that rely on active yeast material or yeast extract, the present invention relies on the use of inactivated yeast biomass. Inactivated yeast biomass refers to yeast cells that have been inactivated through processes like heat treatment or drying, preserving their flavor-enhancing properties while eliminating the need for fermentation or extraction.

[0028] While traditional methods require the metabolic activity of living yeast cells to synthesize and excrete aromatic compounds during fermentation, the use of inactivated yeast biomass bypasses this step entirely. Instead, the preserved yeast cells contain a rich reservoir of flavorenhancing molecules. Manufacturers can harness these compounds to enhance flavor and mask off-notes without the complexities of fermentation or extraction processes. This innovative method provides a convenient and cost-effective solution for improving sensory profiles in meat analogues products, aligning with consumer preferences for natural, clean-label ingredients and simplified production processes.

[0029] The second embodiment of the present invention refers to the use of inactivated yeast biomass for obtaining plant-based food products, preferably plant-based meat, without plant flavour off- notes; or for mitigating or eliminating plant flavour off-notes in plant-based food products, preferably in meat analogues.

[0030] In a preferred embodiment, the present invention refers to the use of inactivated yeast biomass for obtaining plant-based food products further characterized by the retention of added meaty flavour compounds.

[0031] In a preferred embodiment, the yeasts are inactivated through processes like heat treatment or drying.

[0032] In a preferred embodiment, the yeast biomass is added in liquid format.

[0033] Surprisingly, the yeast biomass, when added in liquid format, exhibited the same fiber-like structure observed in both with and without yeast material formulations. The preservation of the fiber-like structure in samples containing liquid yeast biomass suggests a remarkable stability in structural integrity, regardless of the physical state of the yeast. This unexpected outcome presents significant advantages from a sustainability perspective, as it eliminates the need for energy-intensive drying steps during processing. Overall, these results highlight the potential of liquid yeast biomass as a sustainable alternative in food processing, offering structural stability without compromising environmental sustainability.

[0034] In a preferred embodiment, the plant-based food products, preferably plant-based meat, is characterized by a statistically significant increase of Retention Index (RI) 900, 948, 995 and / or 1126 and a statistically significant decreased of RI 1500, 1564, 990, 1714 and / or 1723 measured by gas chromatography using DBWax column [d'Acampora Zellner B, Dugo P, Dugo G, Mondello L. Gas chromatography-olfactometry in food flavour analysis. J Chromatogr A. 2008; 1186(1-2): 123-143. doi:10.1016 / j.chroma.2007.09.006\.

[0035] In a preferred embodiment, the yeast is Crabtree-negative yeast, preferably selected from: Candida utilis Kluyveromyces marxianus, Debaryomyces hansenii or Yarrowia lipolytica. The third embodiment of the present invention refers to plant-based food products, preferably plant-based meat, obtained or obtainable by using or implementing the process of the invention.

[0036] In a preferred embodiment, the plant-based food product, preferably plant-based meat, is characterized by a statistically significant increase of Retention Index (RI) 900, 948, 995 and / or 1126 and a statistically significant decreased of RI 1500, 1564, 990, 1714 and / or 1723 measured by gas chromatography using DBWax column.

[0037] In a preferred embodiment, the plant-based food product, preferably plant-based meat, is further characterized in that it retains poultry-type flavour compounds.

[0038] In a preferred embodiment, meaty flavour compounds are retained throughout processing and storage.

[0039] Kindly note that retention of added meaty flavour can be determined by quantifying the concentration of characteristic volatile markers, such as 2-methylpyrazine, 2,6- dimethylpyrazine, and y-butyrolactone, using headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (HS-SPME-GC-MS). Samples containing inactivated yeast biomass exhibited higher post-processing levels of these compounds relative to control samples, confirming improved retention of added meaty flavour profile.

[0040] Alternative analytical methods commonly known to a person skilled in the art may also be employed to assess flavour retention. These include:

[0041] • Direct thermal desorption GC-MS, which allows quantification of volatile compounds by thermally releasing them from the sample matrix directly into the GC-MS system.

[0042] • Purge and trap GC-MS, which captures volatile compounds in an inert gas stream and concentrates them on a sorbent trap before analysis.

[0043] • Dynamic headspace analysis (DHS), where volatile compounds are purged from the sample under controlled conditions and collected for GC-MS analysis.

[0044] • Stir bar sorptive extraction (SBSE) or solid-phase dynamic extraction (SPDE), as alternative pre-concentration techniques for GC-MS detection.

[0045] • Proton-transfer-reaction mass spectrometry (PTR-MS) or Selected Ion Flow Tube Mass Spectrometry (SIFT-MS), which allow real-time, direct analysis of volatile compounds without chromatographic separation. Sensory evaluation methods, such as quantitative descriptive analysis (QDA), may also complement chemical methods to confirm the retention of poultry-type flavour characteristics.

[0046] Any of these methods may be selected depending on the specific formulation, matrix complexity, and required sensitivity.

[0047] For the purpose of the present invention the following terms are defined:

[0048] • The term "comprising" means including, but not limited to, whatever follows the word "comprising". Thus, use of the term "comprising" indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present.

[0049] • The term "consisting of’ means including, and limited to, whatever follows the phrase “consisting of’. Thus, the phrase "consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present.

[0050] • The expression “plant-based meat” refers to meat produced directly from plants. Instead of relying on an animal to convert plants into meat, meat is more efficiently made by skipping the animal and turning plant ingredients directly into meat. Like animal-based meat, plant-based meat is composed of protein, fat, vitamins, minerals, and water [Stephan van Vliet et al., 2020. Plant-Based Meats, Human Health, and Climate Change. REVIEW article. Front. Sustain. Food Sy st., 06 October 2020. Sec. Agroecology and Ecosystem Services. Volume 4 - 2020 https: / / doi.org / ! 0.3389 / fsufs.2020.00128} .

[0051] • The expression “flavour off-notes” are taints in food products caused by the presence of undesirable compounds. They can originate in raw materials, from chemical changes during food processing and storage, and from micro-organisms.

[0052] • The expression “inactivated yeast biomass” refers to yeast cells that have been inactivated through processes like heat treatment or drying. It consists of yeast cells that are no longer living or active so they cannot produce any of the effects of live yeast, such as fermentation.

[0053] • The expression “yeast biomass in liquid format” just refers to a specific presentation of the yeast biomass. Today, yeast is available in both dried and liquid forms. Liquid format is preferred in the context of the present invention. • The expression “plant-based food product” refers to food products primarily made from a plant protein source. These products comprise ingredients derived from plants, such as legumes (e.g., soy, peas), cereals, nuts, or seeds, which have been processed to form a food matrix that mimics the texture, flavor, nutritional value, or other organoleptic characteristics of animal -derived food products. Representative examples of such food products include meat analogues, such as plant-based burgers, sausages, meatballs, or filets made from texturized vegetable protein or other processed forms of plant protein.

[0054] • “Food Extrusion” is a process where a mix of ingredients is forced through a die or opening using a screw or piston, often while being heated and subjected to high pressure.

[0055] • “Pasteurization” refers to a process which is a comparatively low order of heat treatment, generally at a temperature below the boiling point of water.

[0056] • “Sterilization” refers to a process carried out through a combination of temperature and pressure, is a process that eliminates all living microorganisms, including bacteria, viruses, fungi, and spores, to extend the shelflife and ensure the safety of food products. This is typically accomplished by heating food to temperatures above 100°C (212°F) for a specific duration, often under pressure.

[0057] • "Retention" of a flavour compound, as used herein, refers to the ability of a plantbased food product to preserve or maintain the concentration of one or more volatile compounds associated with poultry-type flavour, such as 2-methylpyrazine, 2,6- dimethylpyrazine, and y-butyrolactone, throughout processing steps including, but not limited to, pasteurisation, sterilisation, and packaging. Retention is quantitatively determined by comparing the concentration of such volatile compounds in the final product with the concentration initially added or present prior to processing, using analytical methods such as HS-SPME-GC-MS, thermal desorption GC-MS, or other techniques known to the skilled person. A higher concentration of the target compounds post-processing indicates greater retention.

[0058] Description of the figures

[0059] Figure 1. Visual appearance of the plant-based meat of the invention.

[0060] Figure 2. Soy-like taste intensity in HME soy samples. Figure 3. Umami taste intensity in HME soy samples.

[0061] Figure 4. Umami taste intensity in HME pea samples.

[0062] Figure 5. Pea-like taste intensity in HME pea samples.

[0063] Figure 6. Umami taste intensity in LME pea samples. Figure 7. Legume taste intensity in LME pea samples.

[0064] Figure 8. Green notes in LME pea samples.

[0065] Figure 9. Cardboard notes in LME pea samples.

[0066] Figure 10. Means of the compounds evaluated in the pasteurized samples with FA1. Different letters in the same compound indicate significant differences (p<0.05). Figure 11. Means of the compounds evaluated in the pasteurized samples with flavorings 1 and 2. Different letters in the same compound indicate significant differences (p<0.05).

[0067] Figure 12. Means of the compounds evaluated in the samples sterilized with FA1. Different letters in the same compound indicate significant differences (p<0.05).

[0068] Figure 13. Means of the compounds evaluated in the samples sterilized with flavorings 1 and 2. Different letters in the same compound indicate significant differences (p<0.05).

[0069] Detailed description of the invention

[0070] The present invention is illustrated by means of the Examples set below, without the intention of limiting its scope of protection.

[0071] Example 1. Materials and methods

[0072] Example 1.1. Materials

[0073] Soybean protein concentrate Arcon® SM Soy Protein Concentrat was obtained from ADM (70% protein). Pea protein isolate NUTRALYS® F85M was obtained from Roquette (83% protein). Saccharomyces cerevisiae yeast SAF INSTANT ROJA was obtained from Lessafre (43% protein). Inactivated dry yeast biomass Kluyveromyces marxianus was produced by MOA FOODTECH (40% protein), named as yeast powder. Inactivated liquid yeast biomass Kluyveromyces marxianus was produced by MOA FOOD TECH (11% dry matter), named as liquid yeast.

[0074] All other chemicals and reagents used for the analysis of odour zones were of analytical grade.

[0075] Example 1.2. Preparation of the extrudates

[0076] Example 1.2.1. High moisture extrusion (HME)

[0077] All extrusion experiments were performed using a FKD 1 ZSK27 doble screw extruder (Cooperion) with ten independent heating zones. Yeast powders were added respectively with Soy protein concentrate at concentrations of 0-60% (shown in Table 1 and Table 2). Yeast powders were added with pea protein isolate at concentrations of 0-60%. The mixed material powder blend was fed to extruder at rate 10 kg / h Water was pumped to the extruder. In one trial the Kmarxianus yeast biomass (Table 3) was incorporated in liquid format with (11% of total dry matter) with a pump directly to the extruder replacing the water pump.

[0078] All trials were operated at screw speed of 480-600 rpm. The heating in the different screw zones were between 40-190°C. The cooling die was then opened for samples to be collected at temperature between 40-85°C. Then, the samples were frozen.

[0079] Table 1. Soy formulas for HME. (DM=Dry matter)

[0080] Table 2. Pea formulas for HME. (DM=Dry matter)

[0081] Table 3. Soy formulas for HME with liquid yeast biomass. (DM=Dry matter)

[0082] Table 4. Pea formulas for HME with liquid yeast biomass. (DM=Dry matter)

[0083] Example 1.2.2. Low moisture extrusion (LME) All extrusion experiments were performed using an LME 1 ZSK27 doble screw extruder (Cooperion) with ten independent heating zones. Yeast powders were added respectively with Soy protein concentrate at concentrations of 0-30%. Yeast powders were added with pea protein isolate at concentrations of 0-30%. The mixed material powder blend was fed to extruder at rate 15 kg / h. Water was pumped to the extruder. All trials were operated at screw speed of 480-600 rpm. The heating in the different screw zones were between 40-190°C. The cooling die was then opened for samples to be collected at temperature between 40-85°C. Then the extrudates were dried at 80°C in an oven.

[0084] Table 5. Soy formulas for LME. (DM=Dry matter)

[0085] Table 6. Pea formulas for LME. (DM=Dry matter)

[0086] Example 1.3. Sensory analysis

[0087] Example 1.3.1. Discrimination sensory test

[0088] A discrimination sensory test was conducted with a tasting panel comprising 12 trained panellists. The formulas selected for this test were HME soyl, HME soy2, HME soy 3, HME_soy4, HME_peal, HME_pea2, HME_pea3 and HME_pea4. The panellists were instructed to evaluate and record the soy-like and pea-like notes, specifically distinguishing between samples containing only soy or pea and those containing soy or pea along with yeast. Additionally, they assessed the umami taste of each sample. The evaluations were carried out using established sensory evaluation techniques, ensuring consistency and reliability in the data collection process.

[0089] Example 1.3.2. Qualitative Descriptive Analysis (QDA) A Qualitative Descriptive Analysis (QDA) was performed to discern differences between samples containing pea alone versus those containing pea along with yeast. The formulas selected for this test were HME_ peal and HME_ pea4. This analysis aimed to establish a detailed sensory profile for each sample and compare the sensory attributes between them. Sixteen expert tasters comprised the panel for this evaluation.

[0090] Example 1.4. Analysis of odour zones of the extrudate

[0091] Formulas HME_ peal and HME_ pea4 were selected to perform the odour zone analysis.

[0092] Example 1.4.1. Dynamic Headspace technique coupled with Solid-Phase Extraction (HS- SPE)

[0093] First step for characterizing aroma profiles is acquisition of representative odor extracts from samples. In this study, we employed a dynamic headspace technique coupled with solid-phase extraction (HS-SPE) to achieve this goal. The optimization process involved determining the appropriate purging time and sample quantity to ensure sufficient aromatic potency Samples, each comprising 100 grams of product, were prepared by cutting the extrudate into 1x1 cm squares and introduced into the device under nitrogen gas flow. The gas stream facilitated the release of volatile compounds from the samples, which were then trapped onto a solid-phase extraction cartridge. Following optimization, the trapped compounds were eluted from the cartridge using ether, and each extract was concentrated to 200 pL for subsequent analysis.

[0094] Example 1.4.2. Gas Chromatography with Olfactometric detection (GC-O-FID)

[0095] For the olfactometric analysis of the extracts to obtain the aroma profile of the samples, we conducted an evaluation to identify the aromatic zones defining them and hierarchize the detected odors in each. The panel comprised six judges, with extensive experience in olfactometric assessment. In the procedure, 3 pL of the concentrated extract were analysed by gas chromatography (Column: DB-WAX) with olfactometric detection and FID detector (GC- O-FID), using a worksheet to record, for each perceived odor, the time, description, and intensity on a scale from 0.5 to 3 points. Data treatment involved calculating the modified frequency (MF) for each odor zone, considering the fraction of panelists detecting the aroma and their attributed intensities, according to Dravnieks' formula. Additionally, the linear retention index (RI), characterizing each odor zone based on its retention time under specific chromatographic conditions, was calculated to further characterize the compounds. Example 2. Material and methods regarding the added flavorings after heat and pressure treatments in plant-based products

[0096] The objective of the validation phase is to analyze the preservative effect of the yeast assayed in the present invention on the added flavorings after heat and pressure treatments in plantbased products.

[0097] Example 2.1 Materials

[0098] Pea protein isolate NUTRALYS® F85M was obtained from ROQUETTE (83-88 %). Inactivated dry yeast biomass Kluyveromyces marxianus was produced by MOA FOODTECH (40 % protein), named as yeast powder. Sunflower oil, native com starch from HELA, flavouring agents “AROMA DE AVE HELA MASTER” (aroma of poultry) (FA1) from Hela and SAVAROME P400 from KERRY (FA2).

[0099] Example 2.2 Preparation of the plant-based products

[0100] The plant-based product is composed of pea protein, sunflower oil, native starch, water, salt, inactivated yeast (0, 2 or 4 %) and two meaty flavoring agents (0 or 1 %), separately or combined. All ingredients are mixed in a blender and distributed into glass containers. Each sample was packaged in a glass container.

[0101] These samples were subjected to pasteurization (80 °C, 30 min) or sterilization treatments (115 °C, 20 min, under pressure) in the STERILVAC 100 autoclave (Daihan Scientific).

[0102] Experimental design:

[0103] Table 7. Samples.

[0104] Past: pasteurization; St: sterilization; FA: flavoring agent.

[0105] Example 2.3 Establish an aroma profile of the sample by GC-O.

[0106] GC-0 profiling was carried out on the 3 chosen samples (SAMPLES 20, 21 and 22, Table 1) to obtain the most relevant odor zones of the sample. Each of the samples was studied by several panelists, obtaining the different odor zones with their aromatic descriptions and chromatographic retention indices on a polar column.

[0107] Example 2.4 Chemical characterization of test samples (GC-MS)

[0108] A protocol based on the solid phase microextraction technique (SPME-GC-MS) coupled with GC-MS analysis was used. Incubation time: 2 min, Extraction time and temperature: 50 min, 40 °C; Sample quantity: 2 ± 0.1 g.

[0109] Example 3. Results

[0110] Example 3.1. Preparation of the extrudates

[0111] Example 3.1.1. High moisture extrusion

[0112] High moisture extrudates (HME) were obtained successfully using a yeast material as a coingredient together with different plant proteins. Tables 8, 9 and 10 represent different yeast material and plant proteins used for the formulation. Respectively it is shown the protein concentration coming from the yeast material in each recipe.

[0113] Through photography, a visual examination of the samples was conducted, focusing on their structural characteristic (see Figure 1 as an example). Overall, the images captured revealed a fiber-like structure present in the formulations with and without yeast material formulations. Surprisingly, the introduction of yeast material did not impact the fiber structure of the samples. This indicates that the addition of yeast does not visibly alter the physical appearance of the product at a macroscopic level. The consistent fiber-like morphology observed across all samples suggests a stable structural integrity, which could have implications for product texture, stability, and overall consumer perception. Further exploration into the impact of yeast incorporation on the macroscopic properties of the formulations may provide valuable insights for product development and optimization. Surprisingly, the yeast biomass, when added in liquid format, exhibited the same fiber-like structure observed in both with and without yeast material formulations. The preservation of the fiber-like structure in samples containing liquid yeast biomass suggests a remarkable stability in structural integrity, regardless of the physical state of the yeast. This unexpected outcome presents significant advantages from a sustainability perspective, as it eliminates the need for energy-intensive drying steps during processing. Overall, these results highlight the potential of liquid yeast biomass as a sustainable alternative in food processing, offering structural stability without compromising environmental sustainability.

[0114] Table 8. Soy HME formula overview. DM: Dry Matter Table 9. Pea HME formula overview. DM: Dry Matter

[0115] Table 10. Soy HME formula with wet biomass overview. DM: Dry Matter Table 11. Pea HME formula with wet biomass overview. DM: Dry Matter

[0116] Example 3.1.2. Low moisture extrusion

[0117] Yeast ingredient was also used as co-ingredient in Low moisture extrudates (LME). Several extrudates were obtained from different plant protein sources with a correct fiber structure.

[0118] Low moisture extrudates (LME) were obtained successfully using a yeast material as a coingredient together with different plant proteins. Table 12 and Table 13 represent different yeast material and plant proteins used for the formulation. Respectively it is shown the protein concentration coming from the yeast material in each recipe. Table 12. Soy LME formula overview. DM: Dry Matter

[0119] Table 13. Pea LME formula overview. DM: Dry Matter

[0120] Example 3.2. Sensory analysis Example 3.2.1. Descriptive sensory test The descriptive sensory test revealed that as the yeast content in the HME (High Moisture Extrusion) formulas increased, there was a reduction in the plant-based protein-like flavor, while simultaneously witnessing an increase in umami flavor.

[0121] The descriptive sensory test revealed that as the yeast content in the LME (Low Moisture Extrusion) formulas increased, there was a reduction in the plant-based ‘ ’legume” notes, ‘’green” notes and ‘’cardboard” notes, while simultaneously witnessing an increase in umami flavor.

[0122] Figures 2-9 provide visual representations of these trends, highlighting the inverse relationship between yeast content and plant-based protein related notes, as well as the positive correlation with umami taste. These results suggest that yeast content plays a crucial role in modulating the sensory characteristics of HME and LME formulas, impacting both flavor profiles. Such insights are invaluable for optimizing product formulations and enhancing overall consumer acceptance.

[0123] Example 3.2.2. Qualitative Descriptive Analysis (QDA)

[0124] Analysis revealed significant differences between the samples in terms of "legume flavor intensity". This intensity notably decreased by approximately 35% with the addition of the yeast biomass to the product. Additionally, reductions were observed in the intensity of "green notes," "undesirable flavors," "rancid flavor," and "cardboard / cardboardy flavor," Table 14 presents the mean values for each descriptor in both samples.

[0125] Table 14. Mean values obtained for each sample and for each attribute assessed.

[0126] Example 3.3. Analysis of odour zones of the extrudate

[0127] The odor zones characterizing the protein extrudate samples were determined using linear retention indices (RI), modified frequency (%MF), and odor descriptors provided by the judges. These data were used to compile a table of odor zones, with the resulting information presented in Table 15.

[0128] Table 15. Odor zones found for the samples using GC-0

[0129] * RI: Retention index. MF: Modified frequency.

[0130] Seventeen distinct odor zones were identified across both samples (although this does not necessarily equate to 17 different compounds, as some odor zones may result from combinations of multiple compounds).

[0131] Table 16 presents the results as differences between %MF observed in the HME pea_4 sample containing yeast biomass minus the %MF in the HMEpea_l sample, ordered by the magnitude of these differences. Three distinct groups of attributes are observed, with some increasing or appearing in the MOA sample, some decreasing or disappearing, and an intermediate group shared between the samples.

[0132] Table 16. MF% differences between descriptors and samples by detected odor zones * RI: Retention index. MF: Modified frequency.

[0133] Further analysis reveals the following key findings:

[0134] Samples primarily differentiate based on increased intensity of RI 900 and RI 995 zones (characterized as sweet, red fruit, strawberry, caramel aromas) in the HMEpea_4 sample, alongside a decrease in RI 990 (green, grass) intensity. Differences are observed for the RI 900, RI 948, and RI 1126 zones, appearing exclusively in the HMEpea_4 sample, as well as an increase observed in the RI 995 zone. These odor zones may explain an increase in sweet notes, either due to their appearance or masking of green notes, this could potentially characterize the HMEpea 4 sample.

[0135] Additionally, the HMEpea 4 sample exhibits a decrease in compounds of RI 1500 and 1564 (green aromas) and a disappearance of unpleasant notes associated with RI 990, 1714, and 1723 zones compared to the HMEpea_l sample. This decrease in notes may relate to the reduction of green notes in HMEpea 1 compared to HMEpea _4, consistent with sensory evaluation results obtained in the QDA.

[0136] Further analysis of %MF values in each sample (Table 17) reveals that, despite similar total intensities (656 versus 584), representing a loss of approximately 10% of the total aroma intensity when using the HMEpea_4 formula versus 100% HMEpea_l, basic differences lie in the significant loss of negative tones, particularly "green" notes (more than 36% loss) and "unpleasant" notes associated with the HMEpea_l sample, and a substantial increase (over 200%) in the more "fruity and sweet" notes characteristic of HMEpea_4. These results align with those found in the QDA, particularly regarding "green" and unpleasant ("legume, green / herbaceous, undesirable, cardboard / cardboard-like") aromas.

[0137] Table 17. Coupled MF% values of the samples

[0138] * l ' _l lo ijied frequency

[0139] Overall, our findings highlight the importance of yeast content as a key determinant of sensory characteristics in HME products. By manipulating yeast levels, manufacturers can tailor flavor profiles to meet consumer preferences and enhance product acceptance.

[0140] Example 4. Results regarding the added flavorings after heat and pressure treatments in plant-based products

[0141] Example 4.1 Aroma profile

[0142] The criterion for choosing the compounds were those that were added with the external aromas (FA1 and / or FA2), that appeared in the olfactometries and for which we had a chemical signal and those that, even without clear identification in the olfactometries, had a chemical signal that allowed us to identify and monitor them in the chosen analysis conditions.

[0143] With this criterion, combining the results obtained in both chemical and sensory studies by GC- O, we arrived at the final choice of the compounds that were followed in the chemical study of the samples in the different processes under study, which are shown in Table 18. Table 18. Definitive list of compounds subject to study in the chemical analysis of the samples.

[0144] Example 4.2 Chemical characterization of test samples (GC-MS) Example 4.2.1 Pasteurization

[0145] Example 4.2.1.1 Pasteurization samples with FA1

[0146] Figure 10 shows the averages of the results obtained for the samples submitted to Pasteurization flavored with FA1.

[0147] Different behaviors are observed in the content of the compounds: • Progressive increase in content when the amount of inactivated yeast increases: 2- methylpyrazine (roasted, meat), 2,6-dimethylpyrazine (toast, roast meat), g- butyrolactone (sweet, creamy), 2-methylbutyric acid (lactic, cheese).

[0148] • Increase with the presence of the inactivated yeast, irrespective of the amount (2% or 4%): isobutyric acid (lactic, cheese) and b -phenyl ethanol (floral). • Increase in the presence of 2% inactivated yeast: acetoin (lactic, bitter), acetic acid (vinegar), acetylfuran, butyric acid (lactic, fatty), 3 -methylbutyric acid (lactic, cheese), hexanoic acid (lactic, fatty).

[0149] • Increased in the presence of inactivated yeast at 4%: 3 -methylbutanal (malty, cured flavour)

[0150] • Not affected by the presence of inactivated yeast: 2-ethylfuran (spicy, smoked) 1-octen- 3-ol (earthy, herbaceous), linalool (floral), guaiacol (woody, spicy), octanoic acid (lactic, fatty) and nonanoic acid (lactic, fatty).

[0151] • Decreased by the presence of inactivated yeast: 2-methylbutanal (malty, cured flavour) and 4-terpineol (floral, herbal).

[0152] Example 4.2.1.1 Pasteurization samples with FA1 and FA2

[0153] Figure 11 shows the averages of the results obtained for the samples subjected to Pasteurisation flavoured with FA1+FA2.

[0154] Different behaviours are observed in the content of the compounds:

[0155] • Progressive increase of the content when the amount of inactivated yeast increases: 2- methylpyrazine (roasted, meat), 2,6-dimethylpyrazine (toast, roast meat), g- butyrolactone (sweet, creamy).

[0156] • Increase with presence of inactivated yeast, irrespective of the amount (2% or 4%): b- phenyl ethanol (floral).

[0157] • Increase in the presence of 2% inactivated yeast: acetoin (lactic, bitter), acetyl furan (caramel, coffee).

[0158] • Increased in the presence of inactivated yeast at 4%: 3 -methylbutanal (Malty, cured flavour).

[0159] • Not affected by the presence of inactivated yeast: 2-methylbutanal (Malty, cured flavour), 2-ethylfuran (spicy, smoked), l-octen-3-ol (earthy, herbaceous), acetic acid, linalool (floral), isobutyric acid (lactic, cheese), 4-terpineol (floral, herbal), butyric acid (lactic, fatty), 2-m ethylbutyric acid (lactic, cheese), 3 -methylbutyric acid (lactic, cheese), hexanoic acid (lactic, fatty), guaiacol (woody, spicy), octanoic acid (lactic, fatty), nonanoic acid (lactic, fatty).

[0160] • Decreased by the presence of inactivated yeast: NON-EXISTING. Example 4.2.2 Sterilization

[0161] Example 4.2.2.1 Sterilization samples with FA1

[0162] Figure 12 shows the averages of the results obtained for the sterilized samples flavored with FA1.

[0163] Different behaviors are observed in the content of the compounds:

[0164] • Progressive increase of the content when the amount of inactivated yeast increases: 2- methylbutanal (Malty, cured flavour), 3 -methylbutanal (Malty, cured flavour), 2- methylpyrazine (roasted, meat), 2,6-dimethylpyrazine (toast, roast meat), acetic acid (vinegar), isobutyric acid (lactic, cheese), g-butyrolactone (sweet, creamy), butyric acid (lactic, fatty), 2-methylbutyric acid (lactic, cheese), b-phenylethanol (floral), nonanoic acid (lactic, fatty).

[0165] • Increase in the presence of inactivated yeast, irrespective of the amount (2% or 4%): octanoic acid (lactic, fatty).

[0166] • Increase in the presence of 2% inactivated yeast: 4-terpineol (floral, herbal).

[0167] • Increased by the presence of inactivated yeast at 4%: acetoin (lactic, bitter), 3- methylbutyric acid (lactic, cheese).

[0168] • Unaffected by the presence of inactivated yeast: l-octen-3-ol (earthy, herbaceous), hexanoic acid (lactic, fatty), guaiacol (woody, spicy).

[0169] • Decreased by the presence of inactivated yeast: 2-ethylfuran, acetyl furan (caramel, coffee), linalool (floral).

[0170] Example 4.2.2.2 Sterilization samples with FA1 and FA2

[0171] Figure 13 shows the averages of the results obtained for the samples submitted to sterilization and flavored with FA11+FA2.

[0172] Different behaviors are observed in the content of the compounds:

[0173] • Progressive increase of the content when the amount of inactivated yeast increases: acetoin (lactic, bitter), 2-methylpyrazine (roasted, meat), 2,6-dimethylpyrazine (toast, roast meat), g-butyrolactone (sweet, creamy), 2-methylbutyric acid (lactic, cheese), b- phenyl ethanol (floral).

[0174] • Increased in the presence of inactivated yeast at 4%: 2-methylbutanal (Malty, cured flavour), 3 -methylbutanal (Malty, cured flavour), acetic acid (vinegar), acetylfuran, isobutyric acid (lactic, cheese), butyric acid (lactic, fatty), 3-methylbutyric acid (lactic, cheese).

[0175] • Lost at 2% inactivated yeast but retained at 4% inactivated yeast: hexanoic acid (lactic, fatty), octanoic acid (lactic, fatty). • Unaffected by the presence of inactivated yeast: l-octen-3-ol (earthy, herbaceous), linalool (floral), 4-terpineol (floral, herbal), guaiacol (woody, spicy), nonanoic acid (lactic, fatty).

[0176] • Decreased by the presence of inactivated yeast: 2-ethylfuran (spicy, smoked).

Claims

CLAIMS1. Process for obtaining plant-based food products without plant flavour off-notes, or with reduced plant flavour off-notes as compared with the plant of origin, which comprises: a. Providing a source of plant protein, b. Mixing the plant protein with yeast biomass, wherein the process is characterized in that the yeast biomass of step b) is inactivated yeast biomass.

2. Process for mitigating or eliminating plant flavour off-notes in plant-based food products which comprises: a. Providing a source of plant protein, b. Mixing the plant protein with yeast biomass, wherein the process is characterized in that the yeast biomass of step b) is inactivated yeast biomass.

3. Process, according to any of the previous claims, characterized in that fermentation or extraction steps are not carried out.

4. Process, according to any of the previous claims, which further comprises subjecting the mixture of step b) to pressure between 0.5 and 50 bar and / or temperature between 60 and 200 °C.

5. Process, according to any of the previous claims, which further comprises extruding a mixture of the plant protein with yeast biomass.

6. Process, according to any of the previous claims, for obtaining plant-based food products further characterized by the retention of meaty flavour compounds.

7. Use of inactivated yeast biomass for obtaining plant-based food products without plant flavour off-notes; or for mitigating or eliminating plant flavour off-notes in plant-based food products.

8. Use, according to claim 7, for obtaining plant-based food products further characterized by the retention of meaty flavour compounds.

9. Process or use, according to any of the previous claims, wherein the yeast is inactivated through processes like heat treatment or drying.

10. Process or use, according to any of the previous claims, characterized in that the yeast biomass is added in liquid format.

11. Process or use, according to any of the previous claims, wherein the plant-based food product is characterized by a statistically significant increase of Retention Index (RI) 900, 948, 995 and / or 1126 and a statistically significant decreased of RI 1500, 1564, 990, 1714 and / or 1723 measured by gas chromatography using DBWax column.

12. Process or use, according to any of the previous claims, wherein the yeast is Crabtreenegative yeast, preferably selected from: Candida mills. Kluyveromyces marxianus, Debaryomyces hansenii or Yarrowia lipolytica.

13. Plant-based food product obtained by the process of claims 1 to 6, or 9 to 12.

14. Plant-based food product, according to claim 13, characterized by a statistically significant increase of Retention Index (RI) 900, 948, 995 and / or 1126 and a statistically significant decreased of RI 1500, 1564, 990, 1714 and / or 1723 measured by gas chromatography using DBWax column.

15. Plant-based food product, according to any of the claims 13 or 14, further characterized in that it retains meaty flavour compounds.

Images