Preparation technology of royal jelly fermented beverage

Abstract

The invention belongs to the technical field of fermentation engineering, and particularly relates to a preparation process of a royal jelly fermented beverage, which comprises the following steps: step 1, taking 200L of fresh milk, sterilizing to kill infectious microbes, and then cooling to 40-45 DEG C; weighing fresh royal jelly accounting for 0.5-1% of the total amount of the raw materials, diluting the fresh royal jelly with fresh milk cooled to 40-45 DEG C, and stirring the diluted royal jelly until the diluted royal jelly is If the early-stage pH value is smaller than 4.5, a buffer system adjusting method in grading treatment can be adopted, the early-stage lactic acid accumulation rate can be reduced through the phosphate buffer effect, and the situation of middle-stage excessive acidification caused by the too high early-stage acidity is avoided; moreover, phosphate promotes the flora to decompose protein and polypeptide in the royal jelly, so that the content of free amino acid in fermentation liquor is increased, meanwhile, peculiar smell substances generated due to abnormal metabolism are reduced, the activity of the fermentation flora is maintained, and the influence of the additive is reduced.

Description

Technical Field

[0001] This invention belongs to the field of fermentation engineering technology, specifically a preparation process for a royal jelly fermented beverage. Background Technology

[0002] Royal jelly fermented beverage is a functional drink made primarily from royal jelly through microbial fermentation. It combines the nutritional properties of royal jelly with the unique flavor and functional components produced during fermentation. The fermentation process includes microencapsulation and segmented fermentation. The segmented fermentation involves first fermenting with lactic acid bacteria for 24 hours, followed by fermentation with yeast for 12 hours. This avoids excessive acidity that inhibits yeast activity while optimizing flavor compound formation. The selection of microbial strains includes single-strain fermentation, such as lactic acid bacteria (producing lactic acid to lower the pH, enhance flavor, and improve stability) and yeast (fermenting ethanol and aromatic substances to give the beverage a unique aroma). Mixed-strain fermentation involves lactic acid bacteria and yeast working synergistically to balance acidity and flavor. Inoculating Lactobacillus bulgaricus with Saccharomyces cerevisiae at a 1:1 ratio can achieve a lactic acid content of 1.5g / 100mL, while also producing esters such as ethyl acetate, enhancing the overall taste.

[0003] However, in the actual fermentation process, the fermentation status of royal jelly beverages needs to be monitored in real time. When the pH of the royal jelly beverage decreases, if only the raw materials are neutralized, the added neutralizing reagents need time to react with the raw materials. When the raw materials are initially neutralized to the adjusted pH value, the internal neutralization process is still in progress. By the time the neutralization adjustment is completed, the pH value of the raw materials has been over-neutralized, causing the originally acidic raw materials to become alkalized. Workers then need to adjust the alkalized raw materials, affecting the fermentation process and reducing the fermentation quality. Furthermore, the sudden change from acidic to alkalized affects the internal microbial community, causing a large number of microorganisms to become inactive or stop growing, resulting in a reduction in the number of fermentation microorganisms in the raw materials, further reducing the fermentation quality. Summary of the Invention

[0004] To overcome the shortcomings of existing technologies and solve the aforementioned technical problems, this invention proposes a preparation process for a royal jelly fermented beverage.

[0005] The technical solution adopted by this invention to solve its technical problem is as follows: This invention proposes a preparation process for a royal jelly fermented beverage, the preparation process steps of which include: Step 1: Take 200L of fresh milk, sterilize it to kill bacteria, and then cool it to 40℃-45℃; weigh out 0.5%-1% of the total raw materials of fresh royal jelly, dilute it with the fresh milk cooled to 40℃-45℃, and stir until it becomes a uniform paste. Step 2: Pour fresh milk and royal jelly into a fermentation container, add auxiliary materials and mix well, then measure the initial pH, and then inoculate with fermentation bacteria at a rate of 2% of the total raw materials; Step 3: Ferment at a constant temperature of 30℃-40℃ for 4-24 hours. When the pH is in the range of 4.2-4.5, inoculate with yeast at a rate of 1% of the total amount and continue fermentation for 8-12 hours. During the fermentation process, check the microbial growth status every 2-4 hours and check the physicochemical indicators every 1-2 hours. Step 4: When the pH of the raw material drops to 4.0-4.5, solidifies into a yogurt-like consistency, and produces a faint milky aroma, stop fermentation; add flavoring, stir well, pasteurize for 20 minutes to kill fermentation bacteria and other microorganisms, and then cool and package.

[0006] Preferably, in step three, based on the physicochemical index results and the acid-base change values ​​during the fermentation cycle of the royal jelly beverage, a graded treatment is adopted, and the microbial growth status is detected after each stage of treatment. The acid-base change values ​​during the fermentation cycle are as follows: 0-4 hours: 5.5-5.2, this period is the adaptation period for lactic acid bacteria; 4-24 hours: 5.2-4.0, this cycle is the period of rapid acid production by lactic acid bacteria, during which lactic acid accumulates; 24-48 hours: 4.0-4.1, this cycle is the stable period of lactic acid bacteria, and the sugar is consumed.

[0007] Preferably, the grading process includes acid treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly acidic, with a pH value between 4.0 and 4.5; Buffer system adjustment method: Add 0.05%-0.1% phosphate to stabilize the pH value at 5.0-5.5 using its buffer pair; Moderately acidic, pH value between 3.5 and 4.0: Alkaline neutralization method: Gradually add food-grade weak alkali to adjust the pH, increasing the amount by 0.05%-0.1% until the pH reaches 5.0-5.5; Severely acidic, pH value less than 3.5: The combined dilution and neutralization treatment method is as follows: First, dilute with a neutral base at a ratio of 1:1 to 1:2 to initially raise the pH value to above 4.0. Then, adjust the pH value to 3.5-4.0 using a moderately acidic method. Finally, adjust the pH value to stabilize at 5.0-5.5 using a mildly acidic method.

[0008] Preferably, the grading treatment includes alkaline treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly alkaline, pH value between 7.0 and 7.5: Method for adjusting acidity: Add food-grade organic acid in a gradient of 0.03%-0.05%, stir evenly, and test the pH value to ensure it stabilizes at 5.0-5.5; Moderately alkaline, pH value between 7.5 and 8.0: Acidification and dilution treatment method: First, add 0.05%-0.1% organic acid to reduce the pH value to 6.5-7.0, then dilute with 10%-20% acidic fruit juice with a pH value of 3.0-3.5, and finally adjust the pH value to be stable at 5.0-5.5; Severely alkaline, pH value greater than 8.0: Reconfiguration.

[0009] Preferably, in step two, the excipients contain 0.5%-3% of a complex of nutrients, including glucose, yeast extract, B vitamins, potassium dihydrogen phosphate, and peptide growth factors.

[0010] Preferably, in step three: If the pH value drops by more than 0.5-1.0 units within 1-2 hours during fermentation, it is considered a sudden drop in pH and is treated using a metabolite inhibition method. If the pH value decreases by less than 0.5 units per hour during a fermentation cycle of more than 4 hours, it is considered to be in a state of slow pH decrease and is treated using a nutrient depletion method.

[0011] Preferably, the metabolite inhibition method includes: Neutralize organic acids: Add edible weak alkali in batches to maintain the pH at 4.0-4.5; To reduce alcohol toxicity: When the alcohol concentration reaches 4%, cool the temperature to 25°C and introduce nitrogen gas, causing the alcohol to evaporate.

[0012] Preferably, the nutrient depletion method includes: Real-time feeding: Add 5% glucose solution via a peristaltic pump at a rate of 1%-2% of the fermentation broth volume to maintain a sugar concentration of 2-3 g / L; Exogenous amino acid supplementation: L-glutamic acid at a concentration of 0.5 g / L and L-cysteine ​​at a concentration of 0.2 g / L are added to promote the synthesis of lactic acid bacteria peptidoglycan. Mineral addition: Add 0.05 g / L of MgSO4 aqueous solution and 0.1 g / L of CaCO3 aqueous solution. The former activates pyruvate kinase and the latter neutralizes lactic acid and provides calcium ions.

[0013] The beneficial effects of this invention are as follows: 1. In the preparation process of the royal jelly fermented beverage described in this invention, if the pH is less than 4.5 in the early stage, a buffer system adjustment method can be adopted in the graded treatment. The buffering effect of phosphate can reduce the rate of lactic acid accumulation in the early stage, avoiding excessive acidification in the middle stage caused by excessive acidity in the early stage. In addition, phosphate promotes the decomposition of proteins and peptides in royal jelly by microorganisms, increases the content of free amino acids in the fermentation liquid, reduces off-flavor substances produced by abnormal metabolism, maintains the activity of fermentation microorganisms, and reduces the influence of additives. Moreover, phosphate can combine with calcium and magnesium ions in royal jelly to form soluble complexes, reduce protein denaturation and precipitation, avoid stratification, and prevent stratification of the growth environment of fermentation microorganisms, which would affect the fermentation quality.

[0014] 2. In the preparation process of the royal jelly fermented beverage described in this invention, during the fermentation of raw materials, the pH value of the raw materials differs from the reference standard. For example, if the measured pH value of the raw materials is lower than the reference standard, it indicates acidification, and workers need to add regulators to increase the pH value. However, the change in pH value takes time, which can easily lead to excessive addition of regulators, resulting in an excessive increase in pH value and affecting the growth of fermentation bacteria. Therefore, a graded treatment method is adopted, based on the difference between the measured pH value and the reference value of the raw materials, to gradually alleviate acidification or alkalization, avoiding sudden changes in pH that could affect the growth of fermentation bacteria. Attached Figure Description

[0015] The invention will now be further described with reference to the accompanying drawings.

[0016] Figure 1 This is a flowchart of the process steps of the present invention; Figure 2 This is a perspective view of the fermentation tank in this invention; Figure 3 This is a schematic diagram of the internal structure of the fermentation tank; Figure 4 This is a cross-sectional view of the sample tube; Figure 5 This is a cross-sectional view of the fermenter from a top-down perspective.

[0017] In the diagram: Fermentation tank 1, transmission device 11, stirring rod 12, sampling tube 13, sampling box 14, filter membrane 15, sample tube 16, sampling hole 17, rotating fan 18, sampling cover plate 19, air pump unit 2. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Example 1: To effectively solve the above problems, see the attached diagram in the instruction manual. Figure 1 As shown, a preparation process for a royal jelly fermented beverage includes the following steps: Step 1: Take 200L of fresh milk, sterilize it to kill any bacteria, and then cool it to 40℃-45℃; weigh out 0.5%-1% of the total raw material weight of fresh royal jelly, dilute it with the cooled fresh milk (40℃-45℃), and stir until it forms a smooth paste; the amount of royal jelly added depends on the type of beverage; the base of the raw material can be fresh milk or a mixture of fresh milk and sterile water; the amount of fresh milk raw material should be adjusted flexibly according to the production volume. Step 2: Pour fresh milk and royal jelly into a fermentation container, add auxiliary materials and mix well, then measure the initial pH, and then inoculate with fermentation bacteria at a rate of 2% of the total raw materials; Step 3: Ferment at a constant temperature of 30℃-40℃ for 4-24 hours. When the pH is in the range of 4.2-4.5, inoculate with yeast at a rate of 1% of the total amount and continue fermentation for 8-12 hours. During the fermentation process, check the microbial growth status every 2-4 hours and check the physicochemical indicators every 1-2 hours. Step 4: When the pH of the raw material drops to 4.0-4.5, it solidifies into a yogurt-like consistency and develops a faint milky aroma, stop fermentation. Add the flavoring agent, stir well, and then pasteurize for 20 minutes to kill fermentation bacteria and other microorganisms. Cool and package. The pH at which fermentation is terminated depends on the fermentation flora and flavor requirements. For example, for lactic acid bacteria fermentation, the pH at termination is 3.5-4.1. In step three, based on the physicochemical index results and the acid-base change values ​​during the fermentation cycle of the royal jelly beverage, a graded treatment was adopted. After each stage of treatment, the microbial growth status was detected. The acid-base change values ​​during the fermentation cycle are as follows: 0-4 hours: 5.5-5.2, this period is the adaptation period for lactic acid bacteria; 4-24 hours: 5.2-4.0, this cycle is the period of rapid acid production by lactic acid bacteria, during which lactic acid accumulates; 24-48 hours: 4.0-4.1, this period is the stable period of lactic acid bacteria, when sugar is consumed; The grading process includes acid treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly acidic, with a pH value between 4.0 and 4.5; Buffer system adjustment method: Add 0.05%-0.1% phosphate to stabilize the pH value at 5.0-5.5 using its buffer pair; Moderately acidic, pH value between 3.5 and 4.0: Alkaline neutralization method: Gradually add food-grade weak alkali to adjust the pH, increasing the amount by 0.05%-0.1% until the pH reaches 5.0-5.5; Severely acidic, pH value less than 3.5: The combined dilution and neutralization treatment method is as follows: First, dilute with a neutral base at a ratio of 1:1 to 1:2 to initially raise the pH value to above 4.0. Then, adjust the pH value to 3.5-4.0 using a moderately acidic method. Finally, adjust the pH value to stabilize at 5.0-5.5 using a slightly acidic method. The graded treatment includes alkaline treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly alkaline, pH value between 7.0 and 7.5: Method for adjusting acidity: Add food-grade organic acid in a gradient of 0.03%-0.05%, stir evenly, and test the pH value to ensure it stabilizes at 5.0-5.5; Moderately alkaline, pH value between 7.5 and 8.0: Acidification and dilution treatment method: First, add 0.05%-0.1% organic acid to reduce the pH value to 6.5-7.0, then dilute with 10%-20% acidic fruit juice with a pH value of 3.0-3.5, and finally adjust the pH value to be stable at 5.0-5.5; Severely alkaline, pH value greater than 8.0: Reconfiguration; Specifically: When monitoring the pH of raw materials, the pH value is determined according to the type of fermentation bacteria inoculated. For example, lactic acid bacteria produce a large amount of lactic acid during fermentation, which makes the pH value of the raw materials lower, and the detection standard value during the fermentation process is also reduced accordingly. During the fermentation process, the pH value of the raw materials may differ from the reference standard. For example, if the measured pH value of the raw materials is lower than the reference standard, it indicates acidification, and workers need to add regulators to raise the pH value. However, the pH value changes over time during the addition process, which can easily lead to excessive addition of regulators, causing an excessive increase in the pH value and affecting the growth of the fermentation microorganisms. Therefore, a graded treatment method is adopted, gradually mitigating acidification or alkalization based on the difference between the measured pH value and the reference value, avoiding sudden changes in pH that could affect the growth of the fermentation microorganisms. Furthermore, treatment can be tailored to different fermentation cycles, such as the following situations: Early stage of fermentation Time range: 0-4 hours after inoculation, depending on the activity of the bacterial strain and temperature; pH range: Initial adjustment value 5.5-5.0, slightly decreasing to 4.5-5.0; Reasons for the change: The strain adapts to the new environment, its metabolic activity is weak, and it only produces a small amount of organic acid. The pH of the raw material is neutral. The buffer substances such as proteins and amino acids in royal jelly temporarily maintain the stability of the pH of the system. For example, if thermophilic streptococci are used for fermentation, the initial pH is adjusted to 5.5, and the reference standard after 4 hours is 4.8-5.2. If the initial pH is less than 4.5, a buffer system adjustment method can be adopted in the staged treatment. The buffering effect of phosphate can reduce the rate of lactic acid accumulation in the early stage, avoiding excessive acidification in the middle stage caused by excessive acidity in the early stage. In addition, phosphate promotes the decomposition of proteins and peptides in royal jelly by the microbial community, increases the content of free amino acids in the fermentation broth, reduces off-flavor substances produced by abnormal metabolism, maintains the activity of the fermentation community, and reduces the impact of additives. Moreover, phosphate can combine with calcium and magnesium ions in royal jelly to form soluble complexes, reduce protein denaturation and precipitation, and avoid stratification, which would lead to stratification of the growth environment of the fermentation community and affect the fermentation quality. Mid-fermentation Time range: Lactic acid bacteria fermentation: 4-24 hours; Yeast fermentation: 24-48 hours; pH range: Lactic acid bacteria fermentation: When the pH drops to 4.0-4.5, lactic acid bacteria produce a large amount of lactic acid; Yeast fermentation: pH drops to 3.5-4.0, accompanied by alcohol production, and less accumulation of organic acids; Reasons for the change: The strains enter the logarithmic growth phase, consuming large amounts of sugar and producing acid; lactic acid bacteria produce lactic acid, while yeast produces a small amount of acetic acid. If the pH level drops below 4.0 within 12 hours, it may indicate excessive bacterial metabolism or contamination by other microorganisms. In such cases, a neutralization method using alkaline substances, such as sodium bicarbonate or calcium carbonate, can be employed during the graded processing. These substances primarily neutralize the organic acids produced during fermentation, maintaining the stability of the system's pH. For instance, when lactic acid bacteria produce lactic acid, causing a decrease in pH, a weak alkali can neutralize the acid and raise the pH level, preventing excessive acidity from inhibiting bacterial activity and thus indirectly alleviating the pH imbalance caused by excessive metabolism. Furthermore, weak alkali itself does not directly inhibit the metabolic activity of the strain, but maintains suitable growth conditions for the strain by adjusting the pH of the environment, reducing the impact of additives on the activity of the microbial community, and avoiding the occurrence of partial uniform inactivation or growth stagnation caused by the addition of additives. By detecting the pH value and using a graded treatment method, the activity of the microbial community can be maintained and the fermentation effect can be improved. If the pH value is below 3.5 during yeast fermentation, it may affect the activity of alcohol dehydrogenase, leading to a decrease in the rate of alcohol production. A combination of dilution and neutralization in the staged treatment can be adopted. The neutral base includes sterile water or skim milk. The addition of the neutral base raises the pH value of the raw material to 4.0. The amount of neutral base added should be adjusted according to the pH value of the raw material, for example, the ratio of neutral base to raw material is 1:1 to 1:2. Later stage of fermentation Time range: Lactic acid bacteria: 24-48 hours; Yeast: 48-72 hours; pH range: Lactic acid bacteria fermentation: 4.0-4.5 until it tends to stabilize; Yeast fermentation: 3.5-4.0 to slowly rise to 3.8-4.2. The rise is due to the production of ammonia by protein hydrolysis, which neutralizes the acid. Reasons for the change: The sugar source is exhausted, the metabolic rate of the strain decreases, the acid production and the system's buffering capacity reach equilibrium, some strains begin to autolyze, releasing alkaline substances such as amino acids, which slightly increases the pH. Fermentation microorganisms thrive in acidic environments, so alkaline environments have a significant impact on them. When the raw materials are slightly alkaline, organic acids should be added in gradients of 0.03%-0.05% of the total raw material to provide a suitable growth environment for lactic acid bacteria and prevent them from becoming inactive or stagnating, rather than simply adjusting the pH. When the raw materials are moderately alkaline, some intolerant fermentation microorganisms may have become inactive or stagnant. The 5%-15% glucose and fructose content in acidic fruit juice can serve as a high-quality carbon source for the fermentation microorganisms, promoting the proliferation of lactic acid bacteria and yeast, and replenishing the number of fermentation microorganisms. Furthermore, the pectin, dietary fiber, and other colloidal substances in the fruit juice can form a network structure with royal jelly proteins, reducing protein precipitation during fermentation, preventing stratification, and improving fermentation quality.

[0020] Example 2: Based on Example 1, in step 2, the excipients contain 0.5%-3% of compound nutrients in total raw materials. The compound nutrients include glucose, yeast extract, compound vitamin B, potassium dihydrogen phosphate and peptide growth factors. Specifically: Nitrogen sources, such as peptone, yeast extract, and amino acids, provide nitrogen for bacterial growth, promote bacterial reproduction and metabolism, and account for 30% of the total complex nutrients; Carbon sources, such as glucose, sucrose, and maltose, serve as energy sources for the microbial community, influencing fermentation speed and product formation, and account for 60% of the total complex nutrients. Vitamins and minerals: B vitamins, inorganic salts, etc., participate in the enzymatic reactions and metabolic regulation of the gut microbiota, and maintain bacterial activity. Complex B vitamins account for 2% of the total complex nutrients, and potassium dihydrogen phosphate accounts for 4% of the total complex nutrients. Growth factors, such as nucleotides and peptides, can promote the growth of specific microbial communities and the synthesis of fermentation products, accounting for 4% of the total complex nutrients; Adding compound nutrients during the formulation design stage provides short-term, fast-acting nutrients for the fermentation microbiota to adapt to the new environment in the early stages, promoting rapid start-up and growth of the fermentation microbiota, reducing the risk of growth stagnation from the source, reducing the occurrence of alkalization, and improving the activity of the fermentation microbiota, thereby improving the fermentation quality of royal jelly beverages. When stagnation occurs, the pH and the number of microorganisms are tested to pinpoint the cause of the problem, and then the raw materials are adjusted in a targeted manner through the above-mentioned graded treatment method to avoid blindly adding materials, which could lead to system imbalance and affect fermentation quality.

[0021] Example 3: Based on Example 2, in Step 3: If the pH value drops by more than 0.5-1.0 units within 1-2 hours during fermentation, it is considered a sudden drop in pH and is treated using a metabolite inhibition method. If the pH value decreases by less than 0.5 units per hour during a fermentation cycle of more than 4 hours, it is considered to be in a state of slow pH decrease and is treated using a nutrient depletion method. The metabolite inhibition method includes: Neutralize organic acids: Add edible weak alkali in batches to maintain the pH at 4.0-4.5; To reduce alcohol toxicity: when the alcohol concentration reaches 4%, cool the temperature to 25°C and introduce nitrogen gas, causing the alcohol to evaporate. The nutrient depletion method includes: Real-time feeding: Add 5% glucose solution via a peristaltic pump at a rate of 1%-2% of the fermentation broth volume to maintain a sugar concentration of 2-3 g / L; Exogenous amino acid supplementation: L-glutamic acid at a concentration of 0.5 g / L and L-cysteine ​​at a concentration of 0.2 g / L are added to promote the synthesis of lactic acid bacteria peptidoglycan. Mineral addition: Add 0.05 g / L of MgSO4 aqueous solution and 0.1 g / L of CaCO3 aqueous solution. The former activates pyruvate kinase and the latter neutralizes lactic acid and provides calcium ions. Specifically: Depletion or imbalance of nutrients Specific manifestations include: stagnant microbial growth during the middle stage of fermentation, no further decrease in acidity test value, and microscopic examination revealing shrunken cells and reduced activity. Cause analysis, for example: Insufficient carbon source: Royal jelly contains about 1.5%-3% natural sugars. Adding too much carbon source may indirectly lead to acidification of the raw materials. If the carbon source is not added enough, it may be depleted during the logarithmic growth phase, and the pH level may not meet the requirements. Nitrogen deficiency: Royal jelly contains about 14%-18% protein, but in the early stage of fermentation, the protease activity is insufficient and the concentration of free amino acids is low, which cannot meet the needs of microbial proliferation. Mineral deficiencies: A lack of magnesium ions can affect the activity of pyruvate dehydrogenase, while a lack of calcium ions can inhibit the synthesis of lactic acid bacteria cell walls. Nutrient depletion-induced growth retardation Carbon source replenishment strategy Real-time feeding method: When the glucose content in the raw material is detected to be <1g / L, a 5% glucose solution is added by peristaltic pump. The amount added can be 1%-2% of the fermentation liquid volume to maintain the sugar concentration in the raw material at 2-3g / L. Nitrogen source activation scheme Exogenous amino acid supplementation: Supplement with L-glutamic acid and L-cysteine ​​aqueous solutions to promote lactic acid bacteria peptidoglycan synthesis. The amount supplemented depends on the amount of fermentation bacteria added; for example, if the amount of fermentation bacteria added is 10g, the supplementation amount is 10L. The amount of amino acids added is determined by the amount of fermentation bacteria added, which can meet the precursor requirements for peptidoglycan synthesis while avoiding the inhibition of cell growth due to increased osmotic pressure caused by excessive concentration. Glutamic acid is a precursor for the synthesis of muramic acid and diaminopimelic acid in peptidoglycan and directly participates in the construction of peptidoglycan monomers. In addition, cysteine, as a sulfur-containing amino acid, enhances the stability of the peptidoglycan layer by participating in the modification of peptide chains during the cross-linking process of peptidoglycan. Mineral Enhancement Adding MgSO4 and CaCO3, the former activates pyruvate kinase, and the latter neutralizes lactic acid and provides calcium ions; During normal fermentation, microorganisms such as lactic acid bacteria produce organic acids through metabolism, causing the pH of the system to decrease at a normal rate, forming an acidic environment. If the pH decreases slowly, it means that the acidic environment of the raw materials is established slowly, which inhibits the growth of lactic acid bacteria and reduces their metabolic activity. Therefore, the compound nutrients added during the raw material preparation can help the fermentation community adapt to the new environment quickly in the early stage of fermentation, establish an acidic environment in advance, avoid inhibiting the growth and reproduction of the fermentation community in the early stage, and ensure that a sufficient number of fermentation community are activated in the early stage of fermentation. Even when nutrients are exhausted in the middle or late stage of fermentation, with the above-mentioned supplementation methods, there can be a sufficient number of fermentation community to meet the loss of community during this process, so that there can still be enough community to continue to grow and reproduce after this process. The combined effect of compound nutrients and nutrient supplementation methods maintains the number of community participating in fermentation and improves the fermentation quality. Furthermore, it can be used in conjunction with a grading process. The grading process addresses the issue of excessive acidification of raw materials, while the above method addresses the problem of insufficient acidification and pH levels not dropping to the reference standard. The two methods work together to improve the fermentation quality of royal jelly beverages. Metabolic product accumulation inhibition Specific manifestations include: a sudden drop in pH to below 3.5, autolysis of the microbial community, and turbidity or off-odor in the fermentation broth; Cause analysis, for example: Lactic acid bacteria fermentation produces lactic acid, and when the concentration is >6g / L, it leads to a decrease in intracellular pH and inhibits the glycolysis pathway. Alcohol toxicity: When the alcohol concentration of yeast fermentation is >5% (v / v), it will damage the lipid bilayer of the cell membrane and inhibit the activity of alcohol dehydrogenase. Add parameters according to the actual situation, for example: Concentration control: The concentration for a single addition should be ≤1.0% (w / v), and it should be prepared as an aqueous solution for use to avoid localized excessive alkalinity; Total addition: Based on 80%-90% of the theoretical acid equivalent for neutralization, with a margin to prevent over-neutralization, the total addition is usually 0.5% to 1.0% (w / v). By adding the raw materials in batches to maintain their pH level, the activity of lactic acid bacteria is ensured while avoiding the risk of alkalization. This ultimately achieves a balance between the number of live bacteria, acid metabolism, and flavor, preventing the impact of a sudden drop in the pH of the raw materials on the fermentation microbiota and improving fermentation quality. Furthermore, in conjunction with a graded treatment method, for example, when the microbiota drops sharply and becomes severely acidic, a weak alkali is added while nitrogen is introduced. Adding a weak alkali to acidified raw materials will produce a reaction that releases carbon dioxide. Carbon dioxide reacts with water to form carbonic acid, which will cause neutralization failure. However, by introducing nitrogen, the carbon dioxide produced in the fermentation container is replaced, reducing the carbon dioxide content and improving the neutralization effect. Furthermore, during yeast fermentation, when the alcohol concentration reaches 4% (v / v), the raw material is cooled to 25°C, and sterilizing air or an inert gas is introduced according to the fermentation time. For example, nitrogen is introduced at 1L / min. Nitrogen accelerates the conversion of ethanol from liquid to gas through a dual mechanism of reducing the partial pressure of ethanol vapor and enhancing the gas-liquid mass transfer efficiency (stirring and bubble action), thereby achieving the volatilization effect, promoting alcohol volatilization, reducing the impact of alcohol on the fermentation process, and improving fermentation quality. Example 4: Based on Embodiment 1, as shown in the accompanying drawings of the specification. Figures 2-5 As shown, the fermentation container used is a conventional fermentation tank 1, and the conventional components inside the fermentation tank 1 include a transmission device 11, a feeding device, a stirring rod 12, and a temperature control device; the fermentation container also includes: Sampling tube 13 is provided on the side wall of fermenter 1 and the sampling tube 13 is evenly distributed in a ring. The sampling tube 13 is set vertically and the top and bottom face the top and bottom of fermenter 1, respectively. The cross-section of the middle part of the sampling tube 13 is trapezoidal and the two sides of the sampling tube 13 near fermenter 1 are inclined towards the center of fermenter 1. A sampling box 14 is disposed inside the sampling tube 13 and is vertically and evenly distributed. A filter membrane 15 is disposed inside the sampling box 14 away from the bottom, and the filter membrane 15 is used to filter yeast. A sample tube 16 is disposed on the side of the sampling tube 13 away from the side wall of the fermenter 1 and is vertically and evenly distributed. One end of the sample tube 16 is close to the center of the fermenter 1, and the other end is located inside the sampling tube 13 and above the sampling box 14. A valve is disposed inside the sample tube 16 and the valve is controlled by the fermenter 1. Sampling holes 17 are evenly distributed at the bottom of the sample tube 16. A rotating fan 18 is fitted onto the sample tube 16, and there is at least one rotating fan 18. The rotating fan 18 is rotatably connected to the sample tube 16 and sealed. One end of the stirring rod 12 is located between adjacent blades in the rotating fan 18 and rotates the rotating fan 18. The outer edge of the blades in the rotating fan 18 is wrapped with solid activated carbon material. Sampling cover plates 19 are evenly provided on the side wall of the fermentation tank 1, and the sampling cover plates 19 correspond to the sampling tube 13. The sampling tube 13 is sealed to the side wall of the fermentation tank 1. An air pump unit 2 is provided on one side of the fermentation tank 1. The air pump unit 2 is connected to the inside of the sampling tube 13 through an air pipe. The air pump unit 2 is a conventional device consisting of a supply air pump, a suction air pump, and a filter device. The air pump unit 2 is used to deliver sterile gas into the sampling tube 13 and to perform negative pressure sampling.

[0022] Specific workflow: During fermentation, raw materials are added to fermentation tank 1, which is kept at a constant temperature by a temperature control device. After preparation, transmission device 11 drives stirring rod 12 to rotate, mixing the raw materials. When the raw materials are stirred, the microorganisms in the raw materials will move towards the side wall of fermentation tank 1 due to centrifugal force, or move along the side wall of fermentation tank 1. The number of microorganisms in the center of fermentation tank 1 decreases, while the number of microorganisms increases in the limited raw materials near the side wall of fermentation tank 1. This results in insufficient nutrients near the side wall to support the growth of the increased number of microorganisms, causing insufficient nutrition, loss of activity, or growth stagnation. Therefore, by setting sampling tubes 13 on the side wall of fermentation tank 1, the raw materials, during rotation, are guided by the inclined side wall of sampling tubes 13, allowing the raw materials carrying microorganisms to move towards the center of fermentation tank 1, dispersing the microorganisms, improving the uniformity of microorganism distribution in the raw materials, and ensuring that the microorganisms have sufficient nutrient supply. Furthermore, since the two sides of the sampling tube 13 are inclined, the force generated when the raw material is guided is smaller than the force generated on the contact plane, which reduces the squeezing force on the bacteria in the raw material, avoids damage to the bacteria during movement, and maintains the viability of the bacteria. When sampling is required, the stirring rod 12 and the raw material are in a static state to avoid the movement of the raw material affecting the distribution of the bacterial community. The air pump unit 2 draws air into the sampling tube 13, and the sampling tube 13 draws the raw material sample through the sampling hole 17 at the bottom of the sample tube 16. The sample flows through the sample tube 16 into the sampling box 14. When the sample just enters the sampling box 14, the filter membrane 15 filters the sample. The filter membrane 15 is a conventional filter membrane used for filtering bacterial communities, separating the liquid and bacterial community in the sample in advance to avoid the bacterial community affecting the detection and improve the detection accuracy. In addition, some bacterial communities will be suspended in the raw material, while others will settle to the lower layer of the raw material due to their weight. Since the sample tubes 16 are vertically and evenly distributed, the sample tubes 16 are divided into... The sampling system is distributed in different layers above, middle, and below the raw materials. By sampling and testing, the stratification of the raw materials is determined, which helps workers determine whether the stratification affects the fermentation quality and improves the monitoring accuracy of the fermentation process. The samples from the upper layer flow and are collected into the upper sampling box 14, and the samples from the lower layer flow and are collected into the lower sampling box 14. Each layer of samples is stored separately. Workers can simply open the sampling cover 19 and take out the sampling box 14 for replacement. The entire process does not require the sampling component to be inserted into the raw materials to take samples, which avoids the risk of contamination. At the same time, the sampling holes 17 are evenly distributed between the side wall and the center of the fermenter 1, which prevents the bacteria that have accumulated on the side wall due to centrifugation from being taken away and affecting the judgment of the distribution of bacteria, thus improving the detection accuracy. After each sampling, raw materials and microorganisms remain on the inner wall of the sample tube 16. Over time, these can deteriorate and produce harmful bacteria, which can contaminate the raw materials and the sample. Therefore, after sampling, the air pump unit 2 pumps gas, such as nitrogen, into the sampling tube 13. The airflow flushes the raw materials remaining inside the sample tube 16 back into the fermenter 1, keeping the sample tube 16 clean and preventing contamination. Furthermore, during fermentation, microorganisms continuously decompose sugars to produce carbon dioxide and water. Carbon dioxide reacts with water to form carbonic acid, which acidifies the raw materials and affects the growth environment of the microbial community. Therefore, by introducing nitrogen gas, which is discharged through sampling hole 17, nitrogen gas is introduced into the liquid in the form of bubbles in the raw materials. A gas-liquid interface is formed on the surface of the bubbles, and liquid carbon dioxide molecules can diffuse into the interior of the bubbles, rise with the nitrogen bubbles and be discharged from the system. This process is equivalent to blowing carbon dioxide out of the liquid with nitrogen gas, similar to blowing away odors in a water glass with air, reducing the risk of acidification of the raw materials and improving the fermentation quality. Moreover, the escape of carbon dioxide from the raw material is limited by the boundary layer formed on the liquid surface, that is, the thin liquid layer near the gas-liquid interface, in which the carbon dioxide concentration gradient is large. The rising motion of nitrogen bubbles stirs the liquid, destroys the boundary layer, and makes it easier for carbon dioxide in the deep liquid to reach the gas-liquid interface and volatilize, thereby improving the discharge effect and further reducing the risk of raw material acidification. In addition, nitrogen bubbles float and stir from bottom to top, bringing bacteria that may have settled in the lower layer to the upper or middle layer. Compared with the stirring action of the stirring rod 12, the movement of bubbles is relatively gentle, avoiding damage to the bacteria and improving the uniformity of the bacteria in the raw materials. When the sample tube 16 is not needed, the control valve of the fermenter 1 is closed, and the raw materials cannot enter the sample tube 16. It can be opened when needed. During the rotation of the stirring rod 12, one end of the branch of the stirring rod 12 rotates between adjacent blades in the rotating fan 18, causing the blades to rotate and achieving the effect of rotating the fan 18. The rotation of the rotating fan 18 pushes the raw material near the side wall of the fermentation tank 1 towards the center of the fermentation tank 1, avoiding the concentration of bacteria around the side wall of the fermentation tank 1 during the stirring process, which would affect the growth and reproduction of the bacteria. Furthermore, if the rotating fan 18 is rotating while nitrogen is being introduced, it pushes the nitrogen bubbles towards the center during the process of pushing the raw material, changing the upward movement trend of the nitrogen bubbles and causing them to move obliquely upward, increasing the range of movement of the nitrogen bubbles, increasing the amount of nitrogen bubbles in contact with carbon dioxide, improving the carbon dioxide discharge efficiency, and reducing the risk of acidification. When the stirring rod 12 moves the edge of the fan blade in the rotating fan 18, slight friction is generated between the two. During the long-term friction process, the solid activated carbon material wrapped around the outer edge of the fan blade will be worn away, and the activated carbon powder will mix into the raw materials. The activated carbon acts as a friction medium to avoid the generation of metal powder from friction, which would contaminate the raw materials. Moreover, the amount of activated carbon that falls off due to long-term wear is small and will not affect the fermentation process of the raw materials. In addition, royal jelly naturally contains a small amount of wax, pigments and biogenic amines. During the fermentation process, off-flavor substances such as volatile sulfur compounds may be produced due to protein decomposition. The porous structure of activated carbon can physically adsorb and trap these substances, while also adsorbing the rancid or bitter components produced by over-fermentation. In the fermentation liquid, the pores of activated carbon bind with off-flavor molecules such as short-chain fatty acids and aldehydes through van der Waals forces, reducing their concentration in the system, thereby improving the flavor of the product, making the fermented royal jelly drink taste milder, reducing the natural fishy smell or the pungent smell produced by fermentation, improving palatability, and thus improving the fermentation quality.

[0023] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A process for the preparation of a fermented royal jelly drink, characterized in that, The preparation process steps include: Step 1: Take 200L of fresh milk, sterilize it to kill bacteria, and then cool it to 40℃-45℃; weigh out 0.5%-1% of the total raw materials of fresh royal jelly, dilute it with the fresh milk cooled to 40℃-45℃, and stir until it becomes a uniform paste. Step 2: Pour fresh milk and royal jelly into a fermentation container, add auxiliary materials and mix well, then measure the initial pH, and then inoculate with fermentation bacteria at a rate of 2% of the total raw materials; Step 3: Ferment at a constant temperature of 30℃-40℃ for 4-24 hours. When the pH is in the range of 4.2-4.5, inoculate with yeast at a rate of 1% of the total amount and continue fermentation for 8-12 hours. During the fermentation process, check the microbial growth status every 2-4 hours and check the physicochemical indicators every 1-2 hours. Step 4: When the pH of the raw material drops to 4.0-4.5, solidifies into a yogurt-like consistency, and produces a faint milky aroma, stop fermentation; add flavoring, stir well, pasteurize for 20 minutes to kill fermentation bacteria and other microorganisms, and then cool and package.

2. The preparation process of fermented royal jelly drink according to claim 1, characterized in that: In step three, based on the physicochemical index results and the acid-base change values ​​during the fermentation cycle of the royal jelly beverage, a graded treatment was adopted. After each stage of treatment, the microbial growth status was detected. The acid-base change values ​​during the fermentation cycle are as follows: 0-4 hours: 5.5-5.2, this period is the adaptation period for lactic acid bacteria; 4-24 hours: 5.2-4.0, this cycle is the period of rapid acid production by lactic acid bacteria, during which lactic acid accumulates; 24-48 hours: 4.0-4.1, this cycle is the stable period of lactic acid bacteria, and the sugar is consumed.

3. The preparation process of fermented royal jelly drink according to claim 2, characterized by: The grading process includes acid treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly acidic, with a pH value between 4.0 and 4.5; Buffer system adjustment method: Add 0.05%-0.1% phosphate to stabilize the pH value at 5.0-5.5 using its buffer pair; Moderately acidic, pH value between 3.5 and 4.0: Alkaline neutralization method: Gradually add food-grade weak alkali to adjust the pH, increasing the amount by 0.05%-0.1% until the pH reaches 5.0-5.5; Severely acidic, pH value less than 3.5: The combined dilution and neutralization treatment method is as follows: First, dilute with a neutral base at a ratio of 1:1 to 1:2 to initially raise the pH value to above 4.

0. Then, adjust the pH value to 3.5-4.0 using a moderately acidic method. Finally, adjust the pH value to stabilize at 5.0-5.5 using a mildly acidic method.

4. The preparation process of fermented royal jelly drink according to claim 2, characterized by: The graded treatment includes alkaline treatment, and the treatment method for the fermentation adaptation period is as follows: Slightly alkaline, pH value between 7.0 and 7.5: Method for adjusting acidity: Add food-grade organic acid in a gradient of 0.03%-0.05%, stir evenly, and test the pH value to ensure it stabilizes at 5.0-5.5; Moderately alkaline, pH value between 7.5 and 8.0: Acidification and dilution treatment method: First, add 0.05%-0.1% organic acid to reduce the pH value to 6.5-7.0, then dilute with 10%-20% acidic fruit juice with a pH value of 3.0-3.5, and finally adjust the pH value to be stable at 5.0-5.5; Severely alkaline, pH value greater than 8.0: Reconfiguration.

5. The preparation process of a royal jelly fermented beverage according to claim 1, characterized in that: In step two, the excipients contain 0.5%-3% of complex nutrients in total raw materials. These complex nutrients include glucose, yeast extract, B complex vitamins, potassium dihydrogen phosphate, and peptide growth factors.

6. The preparation process of a royal jelly fermented beverage according to claim 2, characterized in that: In step three: If the pH value drops by more than 0.5-1.0 units within 1-2 hours during fermentation, it is considered a sudden drop in pH and is treated using a metabolite inhibition method. If the pH value decreases by less than 0.5 units per hour during a fermentation cycle of more than 4 hours, it is considered to be in a state of slow pH decrease and is treated using a nutrient depletion method.

7. The preparation process of a royal jelly fermented beverage according to claim 6, characterized in that: The metabolite inhibition method includes: Neutralize organic acids: Add edible weak alkali in batches to maintain the pH at 4.0-4.5; To reduce alcohol toxicity: When the alcohol concentration reaches 4%, cool the temperature to 25°C and introduce nitrogen gas, causing the alcohol to evaporate.

8. The preparation process of a royal jelly fermented beverage according to claim 6, characterized in that: The nutrient depletion method includes: Real-time feeding: Add 5% glucose solution via a peristaltic pump at a rate of 1%-2% of the fermentation broth volume to maintain a sugar concentration of 2-3 g / L; Exogenous amino acid supplementation: L-glutamic acid at a concentration of 0.5 g / L and L-cysteine ​​at a concentration of 0.2 g / L are added to promote the synthesis of lactic acid bacteria peptidoglycan. Mineral addition: Add 0.05 g / L of MgSO4 aqueous solution and 0.1 g / L of CaCO3 aqueous solution. The former activates pyruvate kinase and the latter neutralizes lactic acid and provides calcium ions.

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