A method for preparing an oat-based beverage by bioenzymatic hydrolysis and products thereof

Abstract

The application discloses a method for preparing an oat-based beverage through biological enzymolysis and a product thereof, and comprises the following steps: continuously hydrolyzing starch, cellulose, protein and other substances in oat slurry by using high-temperature-resistant amylase, cellulase, saccharifying enzyme, papain and alkaline protease, sufficiently releasing reducing substances in the oat, simultaneously generating reducing peptides, improving the free radical scavenging capacity of the oat slurry, and improving the emulsification stability of the oat slurry due to the moderate degradation of macromolecular substances such as starch, cellulose and protein; glycerol is added in three stages at a controlled speed, so that the glycerol is fully mixed with flaxseed oil, and excessive glycerol is avoided from adhering to lipase to reduce the activity of the enzyme, thereby improving the reaction efficiency.

Description

Technical Field

[0001] This invention belongs to the field of plant-based beverages, specifically relating to a method for preparing oat-based beverages through enzymatic hydrolysis and the resulting products. Background Technology

[0002] Oats have a long history of cultivation and abundant yield in my country. They are highly nutritious, containing protein, fat, phenolic compounds, minerals, and beta-glucan, among other nutrients. In 1997, oats were recognized as a functional food by the US FDA, possessing various physiological activities. For example, oat protein typically has a molecular weight of 14-67 kDa and contains all eight essential amino acids, with a nutritional ratio relatively superior to other grains. Similarly, oats are rich in dietary fiber, and its main component, beta-glucan, has multiple benefits, including lowering blood lipid levels, regulating lipid metabolism disorders, and reducing the generation of oxygen free radicals. Oats and oat by-products have been shown to be helpful in treating diabetes and cardiovascular diseases, and oat-based diets have a significant effect on lowering total cholesterol and LDL cholesterol. In recent years, consumer recognition of the nutritional value of oats has led to an increase in demand for oat-based diets.

[0003] Oat milk is a water-soluble extract of oats. While similar in appearance to traditional beverages, oat milk is composed of bioactive compounds that, in addition to nutritional functions, may offer physiological health benefits. The functional component beta-glucan acts as a prebiotic in the gastrointestinal tract, supporting the growth of beneficial microbiota, while the slowly digestible portion of oat starch helps regulate glycemic response. In recent years, oat milk has developed rapidly in my country. Starting in 2018, oat milk began to see niche consumption in China, with various coffee shops and chain restaurants selling it. In 2019, domestic Chinese oat milk brands were established, and numerous more brands emerged in 2020. In 2021, oat milk officially entered a phase of rapid development as a coffee creamer. Due to its low-calorie and high-fiber advantages, consumers have gradually begun to recognize and accept oat milk. The main reason for the booming oat milk industry is the balanced nutritional composition of oats and the health benefits of beta-glucan. As a substitute for animal protein, oat milk can effectively address the allergy problem of lactose intolerance worldwide. Furthermore, oat milk production consumes less energy, contributing to global sustainable development.

[0004] In oat milk processing, the starch in oats easily forms colloids with β-glucan. In addition, starch is also prone to retrogradation during storage, forming clumps. This causes a series of problems such as uneven texture and deteriorated flavor in oat beverages. At the same time, the relatively low oil content in oats affects the taste of oat milk to some extent. Summary of the Invention

[0005] The purpose of this section is to outline some aspects of embodiments of the present invention and to briefly describe some preferred embodiments. Simplifications or omissions may be made in this section, as well as in the abstract and title of this application, to avoid obscuring the purpose of these documents; however, such simplifications or omissions should not be construed as limiting the scope of the invention.

[0006] In view of the problems existing in the above and / or prior art, the present invention is proposed.

[0007] Therefore, the purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing oat-based beverages through enzymatic hydrolysis.

[0008] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0009] After roasting the oat grains, soak them and grind them into a paste;

[0010] After grinding, the pH of the liquid was adjusted to 5.0. After heating, heat-resistant amylase was added for enzymatic hydrolysis. Then, the pH was adjusted to 4.5. Cellulase and saccharifying enzyme were added for enzymatic hydrolysis. Finally, the pH was adjusted to 7.0. After cooling, papain and alkaline protease were added for compound enzymatic hydrolysis. The mixture was then heated and kept warm to inactivate the enzymes. Finally, it was filtered to obtain enzymatically hydrolyzed oat milk.

[0011] Using flaxseed oil and edible glycerol as raw materials, and imprinted lipase as a catalyst, the reaction was carried out in three stages in a continuous manner by adjusting the glycerol injection rate to obtain flaxseed oil diglyceride intermediate product. After removing the lipase, flaxseed oil free fatty acids were added to the intermediate product, and esterification reaction was carried out under high vacuum using Lipase G50 lipase as a catalyst. The free fatty acids in the reaction product were removed by primary molecular distillation to obtain flaxseed oil diglyceride.

[0012] An oat-based beverage is obtained by mixing enzymatically hydrolyzed oat milk, skim milk powder, flaxseed oil diglycerides, white sugar, citric acid, sodium carboxymethyl cellulose, and monoglycerides, followed by high-pressure homogenization, sterilization, and cooling.

[0013] As a preferred embodiment of the preparation method of the present invention, the oat grains are roasted, soaked, and ground into a paste. The roasting conditions are: upper heat 140-160°C, lower heat 120-140°C, time 20-40 min; soaking time 5-10 h; and grinding into a paste at a material-to-liquid ratio of 1:15-25 (g / mL).

[0014] As a preferred embodiment of the preparation method of the present invention, the pH value of the slurry after grinding is adjusted to 5.0, and after heating, a high-temperature resistant amylase is added for enzymatic hydrolysis. The heating temperature is 80-90°C, the amount of high-temperature amylase added is 0.05-0.08%, and the hydrolysis time is 60-90 min.

[0015] In a preferred embodiment of the preparation method described in this invention, the pH value is adjusted to 4.5, and cellulase and saccharifying enzyme are added for enzymatic hydrolysis, wherein the amount of cellulase and saccharifying enzyme added is 0.02% to 0.06%, and the hydrolysis time is 20 to 45 minutes.

[0016] In a preferred embodiment of the preparation method described in this invention, the pH is adjusted to 7.0, and after cooling, papain and alkaline protease are added for compound enzymatic hydrolysis. The amounts of papain and alkaline protease added are 0.1-0.3% and 1-2%, respectively. The cooling temperature is 50-60°C, and the hydrolysis time is 15-45 min.

[0017] As a preferred embodiment of the preparation method described in this invention, the reaction is carried out continuously in three stages by adjusting the injection rate of glycerol. The first stage is to add 5-20% glycerol at a constant rate for 1 hour, the second stage is to add 20-40% glycerol at a constant rate for 1 hour, and the third stage is to add 40-75% glycerol at a constant rate for 0.5 hours. After the addition, the reaction continues for another 5-7 hours.

[0018] As a preferred embodiment of the preparation method described in this invention, the imprinted lipase is prepared by dissolving a nonionic surfactant Tween 20-80 at a concentration of 20-80 mg / L in isopropanol to obtain a mixed solution, adding 10-50% lipase to the solution, stirring the mixture at 400-800 rpm for 15-45 min at 25°C, filtering to obtain the lipase, eluting the lipase with a nonpolar solvent such as n-hexane or octane, filtering, and vacuum drying for 6-10 h to obtain the imprinted lipase; the lipase is a commercially available lipase, including Lipozyme RM IM, NS 40086, and Lipozyme 435; the amount of imprinted lipase added is 8-14 wt%, the molar ratio of glycerol to linseed oil is 2-4:1, and the reaction temperature is 50-70°C.

[0019] As a preferred embodiment of the preparation method described in this invention, the molar ratio of free fatty acids to the glycerol backbone of glycerides in the esterification reaction is 2-4:1, the addition amount is 3-8%, the vacuum degree is 10-30 mbar, and the reaction time is 14-24 h; the yield of flaxseed oil diglycerides is ≥88%, the content of glycidyl esters is <1.5 mg / kg, and the content of chloropropanol esters is <0.4 mg / kg.

[0020] In a preferred embodiment of the preparation method described in this invention, the following components are added: 60-75% enzymatically hydrolyzed oat milk, 8-16% skim milk powder, 8-20% flaxseed oil diglycerides, 5-9% white sugar, 0.04-0.08% citric acid, 0.3-0.6% sodium carboxymethyl cellulose, and 0.5-1% monoglycerides.

[0021] Another objective of this invention is to overcome the shortcomings of the prior art and provide a product obtained by a method for preparing oat-based beverages through bio-enzymatic hydrolysis.

[0022] Beneficial effects of this invention:

[0023] (1) The present invention uses heat-resistant amylase, cellulase, saccharifying enzyme, papain and alkaline protease to continuously hydrolyze starch, cellulose, protein and other substances in oat milk under preferred conditions, thereby fully releasing the reducing substances in oats and generating reducing peptides, thereby enhancing the ability of oat milk to scavenge free radicals. At the same time, due to the moderate degradation of macromolecules such as starch, cellulose and protein, the emulsification stability of oat milk is improved.

[0024] (2) The nonionic surfactant imprinted lipase dissolved in isopropanol in this invention enhances the activity and catalytic stability of the enzyme. According to the properties of the reaction system, glycerol is added in three stages at a controlled rate to ensure that the glycerol is fully mixed with flaxseed oil. At the same time, excessive glycerol adhering to the lipase reduces the activity of the enzyme, thereby improving the reaction efficiency.

[0025] (3) This invention utilizes the characteristic that the lipase Lipase G50 only acts on monoglycerides and diglycerides. By carrying out the esterification reaction under high vacuum and removing fatty acids by primary molecular distillation, the resulting diglycerides have higher purity, higher yield, and lower content of glycidyl esters and chloropropanol esters. Detailed Implementation

[0026] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0027] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0028] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that is mutually exclusive with other embodiments.

[0029] The thermoresistant amylase used in this invention was purchased from Shandong Jinchengda Biotechnology Co., Ltd., with an enzyme activity of 10,000 U / g; the saccharifying enzyme was purchased from Hangzhou Tairen Biotechnology Co., Ltd., with an enzyme activity of 50,000 U / g; the cellulase was purchased from Qingdao Haiweisen Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g; the papain was purchased from Shanghai Xiangqi Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g; and the alkaline protease was purchased from Jiangsu Maisheng Biotechnology Co., Ltd., with an enzyme activity of 50,000 U / g.

[0030] Determination of lipase transesterification activity: The catalytic activity of lipase was evaluated by transesterification between soybean oil and medium-chain triglycerides (MCTs). The substrate was a mixture of soybean oil and MCTs at a molar ratio of 1:1 (soybean oil / MCTs), and the amount of lipase added was 4% relative to the total mass of the substrate. The catalytic reaction was carried out in a round-bottom flask at 50°C and a stirring rate of 600 rpm. After 1 h of reaction, the product was collected to assess its initial activity. The initial activity of the enzyme was assessed based on the content (%) of medium-chain triglycerides after 1 hour of reaction.

[0031] Extraction of polyphenols: Examples 1-3 and Comparative Examples 1-4, after enzymatic hydrolysis, were mixed with a 1% (v / v) HCl-methanol solution at a ratio of 1:25 (m / V) and allowed to stand at 25°C for 24 h. The mixture was then centrifuged at 4000×g for 10 min, and the supernatant was collected. The precipitate was further mixed with a 1% HCl-methanol solution at a ratio of 1:25 and allowed to stand at 25°C for 24 h. The mixture was then centrifuged at 4000×g for 10 min, and the extracts were combined and evaporated to dryness at 40°C. The resulting extract was then dissolved in 3 mL of methanol and stored at -20°C for subsequent analysis.

[0032] Determination of total polyphenol content: Pipette 0.1 mL of appropriately diluted solutions from Examples 1-3 and Comparative Examples 1-4 into a 5 mL test tube. Then, add 0.5 mL of 0.2 mol / L Lolin-phenol reagent and 0.8 mL of 7.5% Na₂CO₃ reagent sequentially, mix thoroughly, and let stand in the dark at 25°C for 30 min before measuring the absorbance at 765 nm. The results are expressed as the total polyphenol content per mL of oat milk equivalent to gallic acid (GAE), in μmol GAE / mL oat milk.

[0033] Determination of DPPH free radical scavenging rate: Mix 500 μL of moderately diluted polyphenol extract with 0.3 mL of 0.6 mmol / L DPPH-ethanol solution, shake well, and react in a 25℃ water bath in the dark for 30 min. Zero the sample with methanol and measure the absorbance at 517 nm. Prepare a standard curve using 0-2000 μmol / L Trolox solution instead of the sample. The results are expressed as the amount of Trolox equivalent to each gram of oat milk (μmol TE / mL oat milk).

[0034] Stability assessment: Accurately weigh a certain amount of sample and place it in a centrifuge tube. Centrifuge at 5000 r / min for 20 min, discard the supernatant, invert the centrifuge tube containing the precipitate for 30 min, accurately weigh the precipitate, and calculate the centrifugation sedimentation rate (SR). Perform three parallel determinations for each sample and take the average value. The lower the centrifugation sedimentation rate, the better the stability of the beverage.

[0035] SR / % = (M1 / M2) × 100

[0036] In the formula: M1 is the mass of the precipitate after centrifugation, g; M2 is the mass of the sample before centrifugation, g.

[0037] Sensory evaluation: Sensory scoring was conducted on a 100-point scale. Ten experienced personnel evaluated the prepared oat milk beverage samples using the scoring criteria shown in Table 1. The beverage was scored based on four aspects: taste, color, aroma, and texture. The sum of the scores for the four sensory evaluations was the total score, which was the sensory index score result of the oat milk beverage.

[0038] Table 1 Sensory Evaluation Criteria for Oat-Based Beverages

[0039]

[0040]

[0041] Example 1

[0042] This invention provides a method for preparing oat-based beverages through enzymatic hydrolysis:

[0043] (1) Preparation of oat milk

[0044] Oat grains were baked in an oven at 150°C (top heat) and 120°C (bottom heat) for 30 minutes. A suitable amount of baked oat grains was weighed and soaked for 5 hours. The oat grains were then ground using a grinder at a ratio of 1:20 (g / mL) and continuously enzymatically hydrolyzed to prepare a highly reducing and stable oat slurry. The pH of the ground slurry was adjusted to 5.0. The temperature was raised to 90°C, and 0.05% thermoresistant amylase was added for 60 minutes of enzymatic hydrolysis. The pH was then adjusted to 4.5, and 0.02% cellulase and 0.02% saccharifying enzyme were added for 45 minutes of enzymatic hydrolysis. The pH was then adjusted to 7.0, and papain and alkaline protease were added for compound enzymatic hydrolysis (0.1% papain and 1% alkaline protease). Hydrolysis was carried out at 50°C for 45 minutes, followed by inactivation at 100°C for 10 minutes. Finally, the slurry was filtered through 100-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0045] Table 2. Polyphenol content, DPPH free radical scavenging rate, and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0046]

[0047]

[0048] (2) Preparation of flaxseed oil diglycerides

[0049] The nonionic surfactant Tween 20 was dissolved in isopropanol at a concentration of 20 mg / L. Isopropanol has better solubility for surfactants and can better disperse them. At the same time, the trace amount of water in isopropanol can keep the enzyme in a suitable soft state. After sufficient dispersion, mixed solution 1 was obtained. Lipozyme RM IM with a mass fraction of 10% was added to mixed solution 1. The mixture was stirred at 400 rpm for 45 min at 25 °C and filtered to obtain lipase. The surfactant imprint template on the lipase Lipozyme RM IM was eluted with the nonpolar solvent n-hexane. The lipase was then filtered and dried in a vacuum desiccator at room temperature for 6 h to remove organic solvents, thus obtaining imprinted lipase.

[0050] The activities of imprinted and non-imprinted lipases are shown below.

[0051] Table 3 Comparison of activities of imprinted and non-imprinted lipases

[0052] Lipase Imprint LipozymeRMIM Non-imprinted LipozymeRMIM Increase in vitality (%) Enzyme activity 34.1％ 22.3％ 52.9

[0053] As shown in Table 3, the activity of Lipozyme RM IM enzyme increased by 52.9% after blotting.

[0054] By adjusting the rate of glycerol injection into the system, the glycerol enzymatic hydrolysis process can be controlled, which facilitates complete mixing of glycerol and triglycerides, while preventing glycerol from adhering to lipase, inhibiting lipase activity, and improving reaction efficiency. Flaxseed oil was added to a batch reactor under nitrogen protection, and imprinted lipase Lipozyme RM IM was added at 8 wt% of the flaxseed oil weight. The temperature was raised to 70°C, and stirring was initiated at 800 rpm. Simultaneously, food-grade glycerol was pumped into the reactor at a molar ratio of glycerol to flaxseed oil of 2:1. The glycerol injection rate was divided into three stages: the first stage involved a uniform injection of 10% glycerol over 1 hour; the second stage involved an injection of 30% glycerol over 1 hour; and the third stage involved an injection of 60% glycerol over 0.5 hours. After the glycerol injection was completed, the reaction continued for 5 hours. The lipase was then filtered to obtain a mixture of glycerides. The resulting glycerol ester mixture was heated to 90°C and stirred slowly at 30 rpm for 1 hour to accelerate the separation of glycerol and glycerol esters. The glycerol was then separated from the glycerol esters, and the recovered glycerol was used as a raw material for a new glycerolysis reaction. The composition of the resulting glycerol esters is shown below.

[0055] Table 4. Composition of glycerides in the reaction products

[0056]

[0057]

[0058] A mixture of glycerides was added to a batch reactor, and free linseed oil fatty acids were added based on the glycerol backbone content of the glycerides, with a molar ratio of free fatty acids to glycerol backbone of glycerides of 2:1. The temperature was raised to 40°C, and stirring was started at 700 rpm. Lipase G50 lipase was added at a rate of 5%, and a vacuum of 20 mbar was applied. The reaction was carried out for 14 hours to obtain the glycerides product. Fatty acids were removed by molecular distillation. The conditions for primary molecular distillation to remove fatty acids were: distillation temperature of 160°C, pressure of 5 Pa, and condenser temperature of 35°C. The obtained product contained 88.2% diglycerides, 0.7% monoglycerides, and 11.1% triglycerides. The yield of diglycerides was 88.2%, the content of glycidyl esters was 1.46 mg / kg, and the content of chloropropanol esters was 0.31 mg / kg.

[0059] (3) Preparation of oat-based beverages

[0060] In the preparation of oat-based beverages, the following additives were used: enzymatically hydrolyzed oat pulp (75%), skim milk powder (8%), flaxseed oil diglycerides (8%), white sugar (8.14%), citric acid (0.06%), sodium carboxymethyl cellulose (0.3%), and monoglycerides (0.5%). The beverages were homogenized three times at 30 MPa and 50 °C using a high-pressure homogenizer. After homogenization, the beverages were sterilized at 121 °C for 15 min to obtain the oat-based beverages. The beverages were then cooled to room temperature for sensory evaluation.

[0061] Example 2

[0062] This invention provides a method for preparing oat-based beverages through enzymatic hydrolysis:

[0063] (1) Preparation of oat milk

[0064] Oat grains were baked in an oven at 140℃ (top heat) and 130℃ (bottom heat) for 40 minutes. A suitable amount of baked oat grains was weighed and soaked for 8 hours. The oat grains were then ground in a grinder at a ratio of 1:15 (g / mL). A high-reducibility, high-stability oat slurry was prepared using a multi-enzyme synergistic continuous hydrolysis process. The pH of the ground slurry was adjusted to 5. The temperature was raised to 80℃, and 0.1% thermoresistant amylase was added. Enzymatic hydrolysis was performed for 90 minutes. The pH was then adjusted to 4.5, and 0.04% cellulase and 0.04% saccharifying enzyme were added. Enzymatic hydrolysis was performed for 30 minutes. The pH was adjusted to 7.0, and papain and alkaline protease were added for compound enzymatic hydrolysis. The amount of papain added was 0.3%, and the amount of alkaline protease added was 1.5%. Enzymatic hydrolysis was performed at 55℃ for 30 minutes. The temperature was then raised to 100℃ and held for 10 minutes to inactivate the enzymes. Finally, the slurry was filtered through 150-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0065] Table 5. Polyphenol content, DPPH free radical scavenging rate, and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0066]

[0067] (2) Preparation of flaxseed oil diglycerides

[0068] The nonionic surfactant Tween 40 was dissolved in isopropanol at a concentration of 40 mg / L. Isopropanol has better solubility for surfactants and can better disperse them. At the same time, the trace amount of water in isopropanol can keep the enzyme in a suitable soft state. After sufficient dispersion, mixed solution 1 was obtained. Lipase NS 40086 at a mass fraction of 30% was added to mixed solution 1. The mixture was stirred at 600 rpm for 30 min at 25 °C and filtered to obtain lipase. The surfactant imprint template on lipase NS 40086 was eluted with the nonpolar solvent octane. The lipase was then filtered and dried in a vacuum desiccator at room temperature for 8 h to remove organic solvents, thus obtaining imprinted lipase.

[0069] The activities of imprinted and non-imprinted lipases are shown below.

[0070] Table 6 Comparison of activities of imprinted and non-imprinted lipases

[0071] Lipase Imprint NS40086 Non-imprinted NS40086 Increase in vitality (%) Enzyme activity 33.1％ 21.6％ 53.2

[0072] As shown in Table 6, the activity of NS 40086 enzyme increased by 53.2% after blotting.

[0073] By adjusting the rate of glycerol injection into the system, the glycerol enzymatic hydrolysis process can be controlled, which facilitates complete mixing of glycerol and triglycerides, while preventing glycerol from adhering to lipase, inhibiting lipase activity, and improving reaction efficiency. Flaxseed oil was added to a batch reactor under nitrogen protection, and imprinted lipase NS 40086 was added at a rate of 14 wt% of the flaxseed oil weight. The temperature was raised to 60°C, and stirring was initiated at 600 rpm. Simultaneously, food-grade glycerol was pumped into the reactor at a molar ratio of glycerol to flaxseed oil of 3:1. The glycerol injection rate was divided into three stages: the first stage involved a uniform injection of 20% glycerol over 1 hour; the second stage involved an injection of 40% glycerol over 1 hour; and the third stage involved an injection of 40% glycerol over 0.5 hours. After the glycerol injection was completed, the reaction continued for 6 hours. The lipase was then filtered to obtain a mixture of glycerides. The resulting glycerol ester mixture was heated to 80°C and stirred slowly at 20 rpm for 1.5 h to accelerate the separation of glycerol and glycerol esters. The glycerol was then separated from the glycerol esters, and the recovered glycerol was used as a raw material for a new glycerolysis reaction. The composition of the resulting glycerol esters is shown below.

[0074] Table 7. Composition of glycerides in the reaction products

[0075] type content(%) monoglycerides 56.6 diglycerides 35.8 Triglycerides 7.6

[0076] A mixture of glycerides was added to a batch reactor, and free linseed oil fatty acids were added based on the glycerol backbone content of the glycerides, with a molar ratio of free fatty acids to glycerol backbone of 3:1. The temperature was raised to 30°C, and stirring was started at 800 rpm. Lipase G50 lipase was added at a rate of 3%, and a vacuum of 10 mbar was applied. The reaction was carried out for 18 hours to obtain the glycerides product. Fatty acids were removed by molecular distillation. The conditions for primary molecular distillation to remove fatty acids were: distillation temperature of 150°C, pressure of 3 Pa, and condenser temperature of 30°C. The obtained product contained 92.1% diglycerides, 0.5% monoglycerides, and 7.4% triglycerides. The yield of diglycerides was 92.1%, the content of glycidyl esters was 1.21 mg / kg, and the content of chloropropanol esters was 0.24 mg / kg.

[0077] (3) Preparation of oat-based beverages

[0078] In the preparation of oat-based beverages, the following additives were used: enzymatically hydrolyzed oat pulp (65%), skim milk powder (16%), flaxseed oil diglycerides (12%), white sugar (5.76%), citric acid (0.04%), sodium carboxymethyl cellulose (0.4%), and monoglycerides (0.8%). The beverage was homogenized three times at 30 MPa and 50 °C using a high-pressure homogenizer. After homogenization, it was sterilized at 121 °C for 15 min to obtain the oat-based beverage. The beverage was then cooled to room temperature for sensory evaluation.

[0079] Example 3

[0080] This invention provides a method for preparing oat-based beverages through enzymatic hydrolysis:

[0081] (1) Preparation of oat milk

[0082] Oat grains were baked in an oven at 160℃ (top heat) and 140℃ (bottom heat) for 20 minutes. A suitable amount of baked oat grains was weighed and soaked for 10 hours. The oat grains were then ground using a grinder at a ratio of 1:25 (g / mL). A high-reducibility, high-stability oat slurry was prepared using a multi-enzyme synergistic continuous hydrolysis process. The pH of the ground slurry was adjusted to 5. The temperature was raised to 85℃, and 0.08% thermoresistant amylase was added for 75 minutes of enzymatic hydrolysis. The pH was then adjusted to 4.5, and 0.06% cellulase and 0.06% saccharifying enzyme were added for 20 minutes of enzymatic hydrolysis. The pH was adjusted to 7.0, and papain and alkaline protease were added for compound enzymatic hydrolysis (0.2% papain and 2% alkaline protease). Hydrolysis was carried out at 60℃ for 15 minutes, followed by inactivation at 100℃ for 10 minutes. Finally, the slurry was filtered through 200-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0083] Table 8. Polyphenol content, DPPH free radical scavenging rate, and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0084]

[0085] (2) Preparation of flaxseed oil diglycerides

[0086] The nonionic surfactant Tween 80 was dissolved in isopropanol at a concentration of 80 mg / L. Isopropanol has better solubility for surfactants and can better disperse them. At the same time, the trace amount of water in isopropanol can keep the enzyme in a suitable soft state. After sufficient dispersion, mixed solution 1 was obtained. Lipozyme 435 with a mass fraction of 50% was added to mixed solution 1. The mixture was stirred at 800 rpm for 15 min at 25 °C and filtered to obtain lipase. The surfactant imprint template on lipase Lipozyme 435 was eluted with the nonpolar solvent n-hexane. The lipase was then filtered and dried in a vacuum desiccator at room temperature for 10 h to remove organic solvents, thus obtaining imprinted lipase.

[0087] The activities of imprinted and non-imprinted lipases are shown below.

[0088] Table 9 Comparison of activities of imprinted and non-imprinted lipases

[0089] Lipase Imprint Lipozyme435 Non-imprinted Lipozyme435 Increase in vitality (%) Enzyme activity 36.2％ 23.1％ 56.7

[0090] As shown in Table 2, the activity of NS 40086 enzyme increased by 56.7% after blotting.

[0091] By adjusting the rate of glycerol injection into the system, the glycerol enzymatic hydrolysis process can be controlled, which facilitates complete mixing of glycerol and triglycerides, while preventing glycerol from adhering to lipase, inhibiting lipase activity, and improving reaction efficiency. Flaxseed oil was added to a batch reactor under nitrogen protection, and imprinted lipase Lipozyme 435 was added at a rate of 18 wt% of the flaxseed oil weight. The temperature was raised to 50°C, and stirring was initiated at 700 rpm. Simultaneously, food-grade glycerol was pumped into the reactor at a molar ratio of glycerol to flaxseed oil of 4:1. The glycerol injection rate was divided into three stages: the first stage involved a uniform injection of 5% glycerol over 1 hour; the second stage involved an injection of 20% glycerol over 1 hour; and the third stage involved an injection of 75% glycerol over 0.5 hours. After the glycerol injection was completed, the reaction continued for 7 hours. The lipase was then filtered to obtain a mixture of glycerides. The resulting glycerol ester mixture was heated to 85°C and stirred slowly at 50 rpm for 0.5 h to accelerate the separation of glycerol and glycerol esters. The glycerol was then separated from the glycerol esters, and the recovered glycerol was used as a raw material for a new glycerolysis reaction. The composition of the resulting glycerol esters is shown below.

[0092] Table 10 Composition of glycerides in the reaction products

[0093] type content(%) monoglycerides 66.4 diglycerides 27.9 Triglycerides 5.7

[0094] A mixture of glycerides was added to a batch reactor, and free linseed oil fatty acids were added based on the glycerol backbone content of the glycerides, with a molar ratio of free fatty acids to glycerol backbone of 4:1. The temperature was raised to 50°C, and stirring was started at 600 rpm. Lipase G50 lipase was added at a rate of 8%, and a vacuum of 30 mbar was applied. The reaction was carried out for 24 hours to obtain the glycerides product. Fatty acids were removed by molecular distillation. The conditions for primary molecular distillation to remove fatty acids were: distillation temperature of 140°C, pressure of 10 Pa, and condenser temperature of 25°C. The obtained product contained 94.1% diglycerides, 0.6% monoglycerides, and 5.3% triglycerides. The yield of diglycerides was 94.1%, the content of glycidyl esters was 0.91 mg / kg, and the content of chloropropanol esters was 0.16 mg / kg.

[0095] (3) Preparation of oat-based beverages

[0096] In the preparation of oat-based beverages, the following additives were used: enzymatically hydrolyzed oat pulp (60%), skim milk powder (13%), flaxseed oil diglycerides (20%), white sugar (5.32%), citric acid (0.08%), sodium carboxymethyl cellulose (0.6%), and monoglycerides (1%). The beverage was homogenized three times at 30 MPa and 50 °C using a high-pressure homogenizer. After homogenization, it was sterilized at 121 °C for 15 min to obtain the oat-based beverage. The beverage was then cooled to room temperature for sensory evaluation.

[0097] The sensory evaluation and stability analysis of Examples 1-3 are shown in Table 11:

[0098] Table 11. Sensory evaluation and stability analysis results of three example products.

[0099] Example 1 Example 2 Example 3 Taste (35 points) 33 32 32 Color (25 points) 24 23 23 Odor (20 points) 18 17 18 Organizational Status (20 points) 18 18 17 Total score 93 90 90 Centrifugal sedimentation rate (%) 0.4 0.5 0.3

[0100] Comparative Example 1

[0101] The difference from Example 1 is that alkaline protease is not used in step (1).

[0102] Oat grains were baked in an oven at 150°C (top heat) and 120°C (bottom heat) for 30 minutes. A suitable amount of baked oat grains was weighed and soaked for 5 hours. The oat grains were then ground using a grinder at a ratio of 1:20 (g / mL) and continuously enzymatically hydrolyzed to prepare a highly reducing and stable oat slurry. The pH of the ground slurry was adjusted to 5.0. The temperature was raised to 90°C, and 0.05% thermoresistant amylase was added for 60 minutes of enzymatic hydrolysis. The pH was then adjusted to 4.5, and 0.02% cellulase and 0.02% saccharifying enzyme were added for 45 minutes of enzymatic hydrolysis. The pH was adjusted to 7.0, and papain (0.1%) was added for compound enzymatic hydrolysis at 50°C for 45 minutes. The temperature was then raised to 100°C and held for 10 minutes to inactivate the enzymes. Finally, the slurry was filtered through 100-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0103] Table 12 Polyphenol content, DPPH free radical scavenging rate and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0104]

[0105] As can be seen from Comparative Example 1, the absence of alkaline protease reduces the polyphenol content in enzymatically hydrolyzed oat milk, reduces the DPPH free radical scavenging capacity of enzymatically hydrolyzed oat milk, and also reduces its stability.

[0106] Comparative Example 2

[0107] The difference from Example 2 is that the pH of the feed solution is modified in step (1) as follows:

[0108] Oat grains were baked in an oven at 140℃ (top heat) and 130℃ (bottom heat) for 40 minutes. A suitable amount of baked oat grains was weighed and soaked for 8 hours. The oat grains were then ground in a grinder at a ratio of 1:15 (g / mL). A high-reducibility, high-stability oat slurry was prepared using a multi-enzyme synergistic continuous hydrolysis process. The pH of the ground slurry was adjusted to 6.0. The temperature was raised to 80℃, and 0.1% thermoresistant amylase was added. Enzymatic hydrolysis was performed for 90 minutes. The pH was then adjusted to 4.0, and 0.04% cellulase and 0.04% saccharifying enzyme were added. Enzymatic hydrolysis was performed for 30 minutes. The pH was adjusted to 8.0, and papain and alkaline protease were added for compound enzymatic hydrolysis. The amount of papain added was 0.3%, and the amount of alkaline protease added was 1.5%. Enzymatic hydrolysis was performed at 55℃ for 30 minutes. The temperature was then raised to 100℃ and held for 10 minutes to inactivate the enzymes. Finally, the slurry was filtered through 150-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0109] Table 13 Polyphenol content, DPPH free radical scavenging rate and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0110]

[0111] Changing the optimal pH value of the enzyme reduced the polyphenol content in the enzymatically hydrolyzed oat milk, decreased its ability to scavenge DPPH free radicals, and also reduced its stability.

[0112] Comparative Example 3

[0113] The difference from Example 2 is that in step (2), the glycerol is added directly to the reaction instead of using the glycerol fractional rate control method.

[0114] By adjusting the rate of glycerol injection into the system, the glycerol enzymatic hydrolysis process can be controlled, facilitating complete mixing of glycerol and triglycerides while preventing glycerol from adhering to lipase, inhibiting lipase activity, and improving reaction efficiency. Flaxseed oil was added to a batch reactor under nitrogen protection, and lipase NS 40086 was added at a rate of 14 wt% of the flaxseed oil weight. The temperature was raised to 60°C, and stirring was initiated at 600 rpm. Simultaneously, food-grade glycerol was pumped into the reactor at a molar ratio of glycerol to flaxseed oil of 3:1. The reaction time was 8.5 h. The lipase was filtered to obtain a mixture of glycerides. The resulting glyceride mixture was heated to 80°C and slowly stirred at 20 rpm for 1.5 h to accelerate the separation of glycerol and glycerides. The glycerol was then separated from the glycerides, and the recovered glycerol was used as a raw material for further glycerol hydrolysis reactions. The composition of the obtained glycerides is shown below.

[0115] Table 14 Composition of glycerides in the reaction products

[0116] type content(%) monoglycerides 45.9 diglycerides 33.5 Triglycerides 20.6

[0117] As shown in the table, the triglyceride content in the Comparative Example 3 system after the reaction was much higher than that in Example 2, indicating that the reaction in Comparative Example 3 was far from reaching equilibrium.

[0118] Comparative Example 4

[0119] The difference from Example 3 is that in step (2), esterification reaction is not carried out by lipase G50, but by primary molecular distillation to remove fatty acids and secondary molecular finishing to remove monoglycerides, thus obtaining diglyceride products.

[0120] Flaxseed oil was added to a batch reactor under nitrogen protection. Lipozyme 435 lipase was added simultaneously at a rate of 18 wt% of the flaxseed oil weight. The temperature was raised to 50°C, and stirring was initiated at 700 rpm. Food-grade glycerol was pumped into the reactor at a molar ratio of 4:1 to flaxseed oil. The glycerol was pumped in three stages: 5% glycerol was pumped in at a uniform rate over 1 hour; 20% glycerol was pumped in over 1 hour; and 75% glycerol was pumped in over 0.5 hours. After the glycerol injection was complete, the reaction continued for 7 hours. The lipase was filtered to obtain a mixture of glycerides. The resulting glyceride mixture was heated to 85°C and slowly stirred at 50 rpm for 0.5 hours to accelerate the separation of glycerol and glycerides. The glycerol was then separated from the glycerides, and the recovered glycerol was used as a raw material for further glycerolysis reactions. The composition of the obtained glycerides is shown below.

[0121] Table 15 Composition of glycerides in the reaction products

[0122] type content(%) monoglycerides 66.4 diglycerides 27.9 Triglycerides 5.7

[0123] Fatty acids were removed by primary molecular distillation, and monoglycerides were removed by secondary molecular distillation. The conditions for primary molecular distillation to remove fatty acids were: distillation temperature 140℃, pressure 10 Pa, and condenser temperature 25℃. After primary molecular distillation, the system contained 65.2% monoglycerides, 28.3% diglycerides, and 6.5% triglycerides. The conditions for secondary molecular distillation to remove monoglycerides were: distillation temperature 200℃, pressure 3 Pa, and condenser temperature 30℃. After secondary molecular distillation, the system contained 1.4% monoglycerides, 80.2% diglycerides, and 18.4% triglycerides. The yield of diglycerides was 28.3%, the content of glycidyl esters was 5.33 mg / kg, and the content of chloropropanol esters was 1.14 mg / kg.

[0124] Comparative Example 5

[0125] The difference from Example 1 is that the pH of the feed solution is modified in step (1) as follows:

[0126] Oat grains were baked in an oven at 150°C (top heat) and 120°C (bottom heat) for 30 minutes. A suitable amount of baked oat grains was weighed and soaked for 5 hours. The oat grains were then ground using a grinder at a ratio of 1:20 (g / mL) and continuously enzymatically hydrolyzed to prepare a highly reducing and stable oat slurry. The pH of the ground slurry was adjusted to 4.0. The temperature was raised to 90°C, and 0.05% thermoresistant amylase was added for 60 minutes of enzymatic hydrolysis. The pH was then adjusted to 5.0, and 0.02% cellulase and 0.02% saccharifying enzyme were added for 45 minutes of enzymatic hydrolysis. The pH was then adjusted to 6.0, and papain and alkaline protease were added for compound enzymatic hydrolysis (0.1% papain and 1% alkaline protease). Hydrolysis was carried out at 50°C for 45 minutes, followed by inactivation at 100°C for 10 minutes. Finally, the slurry was filtered through 100-mesh gauze to obtain the enzymatically hydrolyzed oat slurry for later use.

[0127] Table 16 Polyphenol content, DPPH free radical scavenging rate, and centrifugal sedimentation rate of enzymatically hydrolyzed oat milk

[0128]

[0129] Comparative Example 6

[0130] The difference from Example 1 is that the oat slurry is not enzymatically hydrolyzed after grinding. Other processes are the same as in Example 1. Oat-based beverages are prepared and their stability is determined. The stability results are based on the method for determining centrifugal sedimentation rate.

[0131] Table 17 Comparison of Centrifugal Sedimentation Rate

[0132] Centrifugal sedimentation rate (%) Example 1 0.4 Comparative Example 6 1.3

[0133] Adding high-temperature amylase, cellulase, and saccharifying enzyme can hydrolyze insoluble starch and cellulose in the system, converting them into soluble substances. Adding papain and alkaline protease can hydrolyze proteins into peptides, improving their solubility. At the same time, peptides have certain emulsifying properties, thereby further enhancing their stability.

[0134] Comparative Example 7

[0135] The difference from Example 1 is that no heat-resistant amylase, cellulase, or saccharifying enzyme is added in the preparation of the oat milk. The other processes are the same as in Example 1. Oat-based beverages are prepared and the stability of the oat-based beverages is determined. The stability results are based on the method for determining the centrifugal sedimentation rate.

[0136] Table 18 Comparison of Centrifugal Sedimentation Rate

[0137] Centrifugal sedimentation rate (%) Example 1 0.4 Comparative Example 7 0.8

[0138] The presence of insoluble starch and cellulose in the system negatively impacts its stability, leading to reduced stability of oat-based beverages.

[0139] Comparative Example 8

[0140] The difference from Example 1 is that papain and alkaline protease are not added in the preparation of oat milk. Other processes are the same as in Example 1. Oat-based beverages are prepared and the stability of oat-based beverages is determined. The stability results are based on the method for determining centrifugal sedimentation rate.

[0141] Table 19 Comparison of Centrifugal Sedimentation Rate

[0142] Centrifugal sedimentation rate (%) Example 1 0.4 Comparative Example 8 0.9

[0143] The system contains some insoluble proteins. Without adding proteases to hydrolyze these proteins, the product's stability would be reduced. Furthermore, the release of polyphenols, which bind to the proteins, would decrease their solubility. Hydrolyzed proteins form peptides, which improve solubility. Additionally, some peptides possess emulsifying properties, further enhancing the product's stability.

[0144] Comparative Example 9

[0145] The difference from Example 1 is that flaxseed oil diglyceride is not used; instead, flaxseed oil is used directly. Other processes are the same as in Example 1. Oat-based beverages are prepared, and the stability of the oat-based beverages is determined. The stability results are obtained by referring to the method for determining the centrifugal sedimentation rate.

[0146] Table 20 Comparison of Centrifugal Sedimentation Rate

[0147] Centrifugal sedimentation rate (%) Example 1 0.4 Comparative Example 9 0.9

[0148] Oat-based beverages prepared directly from flaxseed oil have lower stability than those prepared using flaxseed oil diglycerides. Diglycerides possess emulsifying properties and can synergistically enhance the stability of oat-based beverages along with other components in the system.

[0149] Comparative Example 10

[0150] Referring to the blotting conditions of lipase in Example 2, different solvents, including methanol, ethanol, butanol, n-hexane, and octane, were used to dissolve the nonionic surfactant and compare the effects of different solvents on the activity of NS 40086. Other conditions were the same as in Example 2.

[0151] Table 21 Effect of different solvents on the activity of NS 40086

[0152] Solvent type Enzyme activity methanol 15.4％ ethanol 22.7％ Butanol 29.3％ n-Hexane 26.5％ Octane 24.2％ Isopropanol 33.1％ No trace 21.6％

[0153] Comparative Example 11

[0154] Referring to the glycerol hydrolysis conditions of Example 2, nonionic surfactant imprinted NS 40086 and unimprinted NS 40086 were added respectively. The enzyme activity and final enzyme inactivation rate of the two different batches were compared to determine the catalytic stability of the two lipases.

[0155] Table 22 Enzyme activity and final enzyme inactivation rate of NS 40086 in different batches (blotted and non-blotted).

[0156]

[0157]

[0158] Isopropanol has good solubility for surfactants. Furthermore, the trace amounts of water in isopropanol help keep the enzyme in a flexible state, facilitating the action of surfactants on lipases. Since lipases are water-soluble, their surfaces have numerous hydrophilic regions. Adding nonionic surfactants allows these hydrophilic regions to interact with the lipase through hydrophobic interactions—that is, hydrophilic groups bind to hydrophilic groups, and hydrophobic groups bind to hydrophobic groups. After being coated with nonionic surfactants, the lipase surface exhibits greater hydrophobicity, preventing excessive contact between the lipase and polar substances in the system, thus preventing the loss of effective water and improving the structural stability of the lipase, thereby enhancing its catalytic stability. Increased surface hydrophobicity also makes it easier for the lipase to bind to hydrophobic substrates, improving reaction efficiency. Simultaneously, the trace amounts of water in isopropanol keep the enzyme flexible, and the interaction between the nonionic surfactant and the capping of the lipase's active site opens the active site, maintaining its active conformation and further enhancing enzyme activity. The active conformation of the lipase is fixed by drying and dehydration, followed by elution of the imprint template with an organic solvent.

[0159] Nonionic surfactants interact with lipases through hydrophobic forces. These forces are relatively weak and do not alter the conformation of the lipase, nor do they cause conformational changes that lead to its inactivation. Ionic surfactants, on the other hand, interact with lipases through electrostatic forces, causing significant conformational changes and, to some extent, reducing enzyme activity.

[0160] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the present invention.

Claims

1. A method for preparing oat-based beverages via enzymatic hydrolysis, characterized in that: include, After roasting the oat grains, soak them and grind them into a paste; After grinding, the pH of the liquid was adjusted to 5.

0. After heating, heat-resistant amylase was added for enzymatic hydrolysis. Then, the pH was adjusted to 4.

5. Cellulase and saccharifying enzyme were added for enzymatic hydrolysis. Finally, the pH was adjusted to 7.

0. After cooling, papain and alkaline protease were added for compound enzymatic hydrolysis. The mixture was then heated and kept warm to inactivate the enzymes. Finally, it was filtered to obtain enzymatically hydrolyzed oat milk. Using flaxseed oil and edible glycerol as raw materials, and imprinted lipase as a catalyst, the reaction was carried out in three stages in a continuous manner by adjusting the glycerol injection rate to obtain flaxseed oil diglyceride intermediate product. After removing the lipase, flaxseed oil free fatty acids were added to the intermediate product, and esterification reaction was carried out under high vacuum using Lipase G50 lipase as a catalyst. The free fatty acids in the reaction product were removed by primary molecular distillation to obtain flaxseed oil diglyceride. An oat-based beverage is obtained by mixing enzymatically hydrolyzed oat milk, skim milk powder, flaxseed oil diglycerides, white sugar, citric acid, sodium carboxymethyl cellulose, and monoglycerides, followed by high-pressure homogenization, sterilization, and cooling. The preparation process of the imprinted lipase involves dissolving a nonionic surfactant in isopropanol at a concentration of 20-80 mg / L to obtain a mixed solution, adding 10-50% lipase to the solution, stirring the mixture at 400-800 rpm for 15-45 min at 25°C, filtering to obtain the lipase, eluting the lipase with a nonpolar solvent such as n-hexane or octane, filtering, and vacuum drying for 6-10 h to obtain the imprinted lipase. The lipase is a commercially available lipase, including Lipozyme RMIM, NS 40086, and Lipozyme 435. The amount of imprinted lipase added is 8-14 wt%, the molar ratio of glycerol to linseed oil is 2-4:1, and the reaction temperature is 50-70°C. The nonionic surfactant is Tween 20, 40, or 80. The reaction is carried out continuously in three stages by adjusting the injection rate of glycerol. The first stage is to add 5-20% glycerol at a constant rate for 1 hour, the second stage is to add 20-40% glycerol at a constant rate for 1 hour, and the third stage is to add 40-75% glycerol at a constant rate for 0.5 hours. After the addition, the reaction continues for another 5-7 hours.

2. The method as described in claim 1, characterized in that: The process involves baking the oat grains, soaking them, and grinding them into a paste. The baking conditions are: top heat 140~160℃, bottom heat 120~140℃, time 20~40 min; soaking time 5~10 h; and grinding the paste according to a material-to-liquid ratio of 1:15~25 g / mL.

3. The method as described in claim 1, characterized in that: The pH value of the slurry after grinding is adjusted to 5.0, and after heating, a heat-resistant amylase is added for enzymatic hydrolysis. The heating temperature is 80~90℃, the amount of heat-resistant amylase added is 0.05~0.08%, and the enzymatic hydrolysis time is 60~90min.

4. The method as described in claim 1, characterized in that: The pH value is adjusted to 4.5, and cellulase and saccharifying enzyme are added for enzymatic hydrolysis. The amount of cellulase and saccharifying enzyme added is 0.02%~0.06%, and the hydrolysis time is 20~45 minutes.

5. The method as described in claim 1, characterized in that: The pH was adjusted to 7.0, and after cooling, papain and alkaline protease were added for compound enzymatic hydrolysis. The amounts of papain and alkaline protease added were 0.1-0.3% and 1-2%, respectively. The cooling temperature was 50-60℃, and the hydrolysis time was 15-45 min.

6. The method as described in claim 1, characterized in that: In the esterification reaction, the molar ratio of free fatty acids to the glycerol backbone of glycerides is 2-4:1, the addition amount is 3-8%, the vacuum degree is 10-30 mbar, and the reaction time is 14-24 h; the yield of flaxseed oil diglycerides is ≥88%, the content of glycidyl esters is <1.5 mg / kg, and the content of chloropropanol esters is <0.4 mg / kg.

7. The method as described in claim 1, characterized in that: The amount of enzymatically hydrolyzed oat milk added is 60-75%, the amount of skim milk powder added is 8-16%, the amount of flaxseed oil diglycerides is 8-20%, the amount of white sugar added is 5-9%, the amount of citric acid added is 0.04-0.08%, the amount of sodium carboxymethyl cellulose added is 0.3-0.6%, and the amount of monoglycerides added is 0.5-1%.

8. An oat-based beverage prepared by the method according to any one of claims 1 to 7.

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