Edible compound enzyme as well as preparation method and application thereof

Abstract

The invention discloses an edible compound enzyme as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing fresh leaves of moringa oleifera and fresh leaves of lithocarpus litseifolius, and then carrying out composite enzymolysis pretreatment; then adding malt extract as a fermentation accelerant, and sequentially carrying out high-frequency sound wave fermentation and low-frequency sound wave auxiliary fermentation; and finally, carrying out sterilization and low-temperature concentration on the fermentation liquor to obtain the edible compound enzyme. According to the method, through cooperation of enzymolysis, a specific fermentation accelerant and two-stage variable-frequency sound wave auxiliary fermentation, characteristic active ingredients such as moringa oleifera flavone and Lithocarpus litseifolius fresh leaf dihydrochalcone (such as phlorizin) can be efficiently enriched, and the fermentation activity of an enzyme product is remarkably improved. The enzyme fermentation stock solution prepared by the method can be directly diluted with warm water for drinking, is harmonious in taste and rich in nutrition, and meets the requirements of ready-to-eat enzyme products; meanwhile, the compound can also be used as a functional fermentation raw material to be applied to baking food.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of food bioengineering, and particularly relates to a kind of edible compound enzyme and its preparation method and application. BACKGROUND

[0002] Enzyme, as an active product prepared by microbial fermentation using plants and other materials as raw materials, is rich in enzymes, organic acids, vitamins and plant secondary metabolites, and is considered to have many health benefits such as promoting digestion and regulating intestinal flora, and is an important category in the field of healthy food. Moringa oleifera leaf and Lithocarpus polystachyus leaf are both medicinal and edible plants with high nutritional value. Developing them into edible compound enzyme products is expected to achieve the superposition of nutrition and function, and has a broad market prospect.

[0003] However, the existing technology in this direction has obvious path dependence and limitations. For example, the existing Chinese patent CN104957610A discloses a preparation method of Moringa oleifera and Maca edible compound enzyme. The typical operation is to crush the raw materials and inoculate mixed strains for one-time static fermentation. The core purpose of this method is only to obtain a terminal enzyme product that can be directly consumed. The process design is rough, and the directional enrichment efficiency of Moringa oleifera flavones and Lithocarpus polystachyus characteristic active ingredients is limited. More importantly, this technical idea does not consider whether the prepared enzyme has the potential for secondary application as a ferment. The fermentation activity of the microorganisms contained therein has not been optimized and cannot be directly used in the fermentation process of bread and other foods that require strong gas production capacity. On the other hand, in order to improve the quality of bread, various chemical improvers or natural ferments (such as sourdough) have been developed in the professional baking field. However, the core of these technologies is to optimize the flour system and the fermentation performance of yeast. The ingredients (such as various enzyme preparations, emulsifiers) or processes (such as the preparation of a sourdough of specific lactic acid bacteria) are different from the enzyme preparation system aimed at enriching plant active ingredients. Their goal is to improve the specific volume, texture or flavor of bread, rather than to create a compound functional fermentation raw material that is rich in plant active ingredients and has high fermentation activity. This disconnection between "function" and "performance" leads to a lack of a compound product that can be used as a high-value functional food and as a high-efficiency natural ferment.

[0004] Therefore, an innovative preparation method is urgently needed to break through the bottleneck of existing technology and develop a new edible enzyme raw material with high active ingredient content and high fermentation activity. SUMMARY

[0005] In view of the above, the present invention provides an edible compound enzyme prepared by compounding raw materials, compound enzymatic hydrolysis, two-stage sonic-assisted fermentation and adding malt extract. This enzyme also has high content of characteristic active ingredients such as moringa flavonoids and litsea cubeba dihydrochalcones. It can be directly diluted and consumed, or used as a functional fermentation raw material in baked goods.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A method for preparing an edible compound enzyme includes the following steps: S1. Compound enzymatic hydrolysis pretreatment: Mix fresh Moringa leaves and fresh Litsea cubeba leaves, crush them, add honey and small molecule water, mix evenly, and then add a compound enzyme solution containing cellulase and pectinase for enzymatic hydrolysis. S2. Two-stage acoustic-assisted fermentation: A compound fermentation agent is inoculated into the material treated in S1 and fermentation is carried out in two stages: A. First stage: Fermentation is carried out with lactic acid bacteria as the main component under the assisted fermentation of sound waves at a frequency of 7500-8500Hz; B. Second stage: Fermentation is carried out with yeast as the main component under the assisted fermentation of sound waves at a frequency of 1000-2000Hz; S3. Activity Stabilization: After fermentation, the fermentation broth is sterilized and concentrated at low temperature to obtain the edible compound enzyme. In the initial stage of S1 or S2, a fermentation promoter accounting for 5%-15% of the total mass of the raw materials is added, and the fermentation promoter is malt extract.

[0007] Preferably, in the compound enzymatic hydrolysis pretreatment, the compound enzyme further includes a protease, the enzymatic hydrolysis temperature is 45-55℃, and the enzymatic hydrolysis time is 1-3 hours.

[0008] Preferably, in the compound enzymatic hydrolysis pretreatment, the amount of cellulase added is 0.5%-2% of the total mass of the raw materials, the amount of pectinase added is 0.2%-1% of the total mass of the raw materials, and the amount of protease added is 0.1%-0.5% of the total mass of the raw materials.

[0009] Preferably, the lactic acid bacteria inoculated in the first stage is Lactobacillus plantarum, and the inoculation amount is 1%-5% (v / v) of the total volume of the fermentation substrate; the yeast inoculated in the second stage is Saccharomyces cerevisiae, and the inoculation amount is 0.5%-3% (v / v) of the total volume of the fermentation substrate.

[0010] Preferably, the fermentation time of the first stage is 24-36 hours, and the fermentation of the second stage continues until the pH value of the fermentation broth stabilizes at 3.8-4.2.

[0011] Preferably, the sterilization is performed by filtration sterilization using a 0.22μm microporous filter membrane.

[0012] Another objective of this invention is to provide an edible compound enzyme prepared by the method described above. The enzyme fermentation stock solution prepared by the above method can be directly diluted with warm water for drinking, with a harmonious taste and rich nutrition, meeting the requirements of ready-to-eat enzyme products; at the same time, it can also be used as a functional fermentation raw material in baked goods.

[0013] Another object of the present invention is to provide a method for preparing bread, wherein the above-mentioned edible compound enzyme is used in the preparation process.

[0014] Preferably, the edible compound enzyme is used as a leavening agent in the bread fermentation process, that is, as the sole leavening agent.

[0015] Preferably, the edible compound enzyme and commercial yeast are used together as leavening agents in the bread fermentation process, and the mass ratio of the edible compound enzyme to the commercial yeast is 1:1 to 1:2.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: 1. This invention proposes a synergistic effect of "compound enzymatic hydrolysis" and "two-stage variable frequency acoustic wave-assisted fermentation" to achieve efficient and targeted enrichment of characteristic active ingredients of Moringa flavonoids and Litsea cubeba dihydrochalcone (such as phlorizin). Traditional single fermentation processes do not completely break down cell walls, resulting in low component release efficiency. Specifically, this invention first uses a compound enzyme system containing cellulase, pectinase, etc., to specifically hydrolyze the cell walls of fresh Moringa and Litsea cubeba leaves, fully releasing intracellular active substances. Then, through the time-sequential regulation of "high-frequency acoustic waves promoting rapid acid building by lactic acid bacteria and inhibiting other bacteria" and "low-frequency acoustic waves guiding yeast aroma production and secondary metabolism," an optimal metabolic environment is created for microbial transformation of the aforementioned precursor substances. Experimental data show that this synergistic process results in the content of characteristic components in the final product being more than three times that of traditional static fermentation processes, and is significantly superior to single-frequency acoustic wave fermentation.

[0017] 2. This invention proposes to significantly enhance the dynamic fermentation activity of malt extract as a natural leavening agent by adding a specific fermentation promoter—malt extract—with almost no impact on the content of the final characteristic components of the enzyme. Furthermore, in this invention, malt extract does not simply act as a carbon source; its rich content of vitamins, amino acids, and other growth factors creates a unique synergy with the specific process system, enabling targeted activation and nourishment of fermenting microorganisms. Comparative experiments have confirmed that the fermentation speed of the dough prepared with added malt extract is approximately 53% higher than that of the unadded group, and this improvement is significantly better than the control group using an equal amount of corn extract. This successfully solves the industry problem of insufficient fermentation efficiency commonly found in highly active edible compound enzymes.

[0018] 3. The high-quality enzyme prepared by the above method is a "dual-function" natural leavening agent with both high active ingredients and high fermentation activity, exhibiting superior performance in baked goods such as bread. This enzyme can not only independently complete bread making as the sole leavening agent, imparting a unique plant aroma and superior anti-aging properties to the product; it can also synergize with commercial yeast, demonstrating comprehensive advantages such as shorter fermentation time, larger bread volume, and better texture when used in combination (e.g., a 1:1 mass ratio). Furthermore, the functional components (such as flavonoids) in this invention's enzyme are resistant to the baking process and can be transferred to the final food product, realizing a value upgrade from "edible enzyme" to "functional fermentation raw material," providing innovative raw materials and healthy consumption options for developing nutritionally fortified baked goods.

[0019] In summary, this invention not only provides a method for the stable production of edible compound enzymes with ultra-high activity, but also creatively endows the enzyme with excellent natural fermentation agent properties. At the same time, it solves the technical contradiction of the difficulty in achieving both "ingredients" and "efficacy" in traditional processes, and has outstanding industrial application value. Detailed Implementation

[0020] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention are described in detail below. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0021] The raw materials and equipment used in the following embodiments are as follows: Ingredients: fresh Moringa leaves (washed, drained, and crushed to pass through an 80-mesh sieve), fresh Litsea cubeba leaves (washed, drained, crushed, and passed through an 80-mesh sieve), small molecule water (pH 6.5-7.0, conductivity <10 μS / cm), malt extract (a food-grade powder rich in maltose and other components, made by soaking and drying malted barley), and honey (commercially available).

[0022] Enzyme preparations: food-grade cellulase (enzyme activity ≥100,000 U / g), pectinase (enzyme activity ≥50,000 U / g), and protease (enzyme activity ≥50,000 U / g), purchased from commercially available sources.

[0023] Fermentation inoculum: Lactobacillus plantarum (live count ≥ 1 × 10⁻⁶) 11 CFU / g), brewer's yeast (live count ≥1×10⁻⁶) 10 CFU / g), purchased from the market.

[0024] Main equipment: constant temperature incubator, program-controlled acoustic fermentation device (frequency adjustable), low temperature spray dryer, microporous filtration system.

[0025] Example 1: This embodiment proposes a method for preparing an edible compound enzyme, including the following steps: (1) Pretreatment with compound enzymatic hydrolysis: Take 160g of fresh Moringa leaves and 40g of fresh Litsea cubeba leaves, mix them, crush them, add 30g of honey and 500ml of small molecule water, and stir evenly. Then add compound enzyme solution, in which the amount of cellulase added is 0.4% of the total mass of raw materials and the amount of pectinase added is 0.2%. Adjust the pH of the system to 5.5 with buffer solution, and enzymatically hydrolyze at 45℃ for 3 hours.

[0026] (2) Add fermentation promoter: Add malt extract, the amount of which is 5% of the total mass of raw materials.

[0027] (3) Two-stage acoustic-assisted fermentation: Lactobacillus plantarum was inoculated into the enzymatically hydrolyzed material at an inoculation rate of 3% (v / v) of the total fermentation substrate volume. Fermentation was carried out for 24 hours at 30℃ and 7500Hz with acoustic assistance. Subsequently, Saccharomyces cerevisiae was inoculated at an inoculation rate of 0.5% (v / v) of the total fermentation substrate volume, and the acoustic frequency was adjusted to 2000Hz. Fermentation continued until the pH of the fermentation broth stabilized at 4.2.

[0028] (4) Activity stabilization: The fermentation broth was filtered and sterilized through a 0.22μm microporous membrane. The resulting filtrate was concentrated by low-temperature spray drying (inlet air temperature ≤60℃, outlet air temperature ≤45℃) to obtain edible compound enzyme powder, which was denoted as sample S1.

[0029] Example 2: This embodiment proposes a method for preparing an edible compound enzyme, including the following steps: (1) Pretreatment with compound enzymatic hydrolysis: Take 150g of fresh Moringa leaves and 50g of fresh Litsea cubeba leaves, mix them, crush them, add 40g of honey and 600ml of small molecule water, and mix well. Then add compound enzyme solution, in which the amount of cellulase added is 0.75% of the total mass of raw materials, the amount of pectinase added is 0.25%, and the amount of protease added is 0.1%. Adjust the pH of the system to 5.0 with buffer solution, and enzymatically hydrolyze at 50℃ for 2 hours.

[0030] (2) Add fermentation promoter: Add malt extract at a rate of 10% of the total mass of raw materials.

[0031] (3) Two-stage acoustic-assisted fermentation: 2% (v / v) Lactobacillus plantarum was inoculated and fermented for 30 hours under acoustic-assisted fermentation at 30℃ and 8000Hz. Then, 1% (v / v) Saccharomyces cerevisiae was inoculated, the acoustic frequency was adjusted to 1500Hz, and fermentation continued until the pH value stabilized at 3.9.

[0032] (4) Activity stabilization: Same as in Example 1, enzyme powder sample S2 was obtained.

[0033] Example 3: (1) Pretreatment with compound enzymatic hydrolysis: Take 140g of fresh Moringa leaves and 60g of fresh Litsea cubeba leaves, mix them, crush them, add 50g of brown sugar and 700ml of small molecule water, and mix well. Then add compound enzyme solution, in which the amount of cellulase added is 2% of the total mass of raw materials, the amount of pectinase added is 1%, and the amount of protease added is 0.5%. Adjust the pH of the system to 4.5 with buffer solution, and enzymatically hydrolyze at 55℃ for 1 hour.

[0034] (2) Add fermentation promoter: Add malt extract, the amount of which is 15% of the total mass of raw materials.

[0035] (3) Two-stage acoustic-assisted fermentation: 1% (v / v) Lactobacillus plantarum was inoculated and fermented for 36 hours under acoustic-assisted fermentation at 30℃ and 8500Hz frequency. Then, 3% (v / v) Saccharomyces cerevisiae was inoculated, the acoustic frequency was adjusted to 1000Hz, and fermentation continued until the pH value stabilized at 3.8.

[0036] (4) Activity stabilization: Same as in Example 1, enzyme powder sample S3 was obtained.

[0037] To clearly and in isolation verify the non-obviousness of the various technical features of this invention and their synergistic contributions, the following comparative examples are specifically established: Comparative Example D1 (Traditional Static Fermentation): After the raw materials are mixed, no enzymatic hydrolysis is performed. Lactic acid bacteria and yeast are inoculated at the same time, and the mixture is statically fermented at 30°C without any specific sound wave intervention.

[0038] Comparative Example D2 (without compound enzymatic hydrolysis): without the compound enzymatic hydrolysis pretreatment in step 1, 10% malt extract was added directly and the same two-stage variable frequency acoustic wave assisted fermentation as in Example 2 was carried out.

[0039] Comparative Example D3 (without malt extract): The same complex enzymatic hydrolysis and two-stage variable frequency sonic assisted fermentation as in Example 2 were performed, but without the addition of malt extract.

[0040] Comparative Example D4 (single high-frequency sound control): The same complex enzymatic hydrolysis as in Example 2 was performed with the addition of 10% malt extract, but the entire fermentation process (two stages) was assisted by 8000Hz high-frequency sound waves.

[0041] Comparative Example D5 (single low-frequency sound control): The same compound enzymatic hydrolysis as in Example 2 was performed with the addition of 10% malt extract, but 1500Hz low-frequency sound waves were used to assist fermentation throughout the entire fermentation process (two stages).

[0042] Comparative Example D6 (replacing malt extract): The same complex enzymatic hydrolysis and two-stage variable frequency acoustic wave-assisted fermentation as in Example 2 were performed, but corn extract (10% of the total raw material mass) was used instead of malt extract. The corn extract is a powdered product obtained from corn starch through enzymatic hydrolysis, purification, and spray drying; its main components are fermentable sugars such as maltose and glucose.

[0043] Experimental Example 1: The enzyme products obtained in Examples 1-3 of this invention were tested for key indicators to prove that they not only fully comply with industry standards, but also significantly surpass them in core indicators, demonstrating the fundamental superiority of the method of this invention.

[0044] Table 1: Comparison of key indicators of the enzyme product of this invention with industry standard (QB / T 5323-2018) According to the data in Table 1, the enzyme products prepared in the three embodiments of this invention fully meet and exceed the requirements of the industry standard "Edible Compound Enzymes" (QB / T 5323-2018) in all key indicators, including pH value, total acid, polyphenols, number of active microorganisms, and SOD enzyme activity. This indicates that, within the parameter range defined by this invention, high-quality edible compound enzymes can be stably and reliably produced, providing qualified and high-quality basic raw materials for subsequent functional applications.

[0045] Experimental Example 2: The purpose of this experiment is to reveal the independent contributions and synergistic effects of the two core features, "compound enzymatic hydrolysis" and "two-stage variable frequency sound control," in the enrichment of Moringa flavonoids and characteristic dihydrochalcones (calculated as phlorizin) of Litsea cubeba. A comparison was made with Example 2 (S2) as a baseline, and with various comparative examples.

[0046] Table 2: Effect of different preparation processes on the enrichment efficiency of characteristic active ingredients According to the data in Table 2, the characteristic component content of Comparative Example D2 (without enzymatic hydrolysis) was higher than that of the traditional process D1, but significantly lower than that of Example 2. This directly proves that the "compound enzymatic hydrolysis pretreatment" is a fundamental and crucial step in effectively breaking down plant cell walls and releasing intracellular Moringa flavonoids and characteristic dihydrochalcones of Litsea cubeba. The characteristic component content of Comparative Examples D4 and D5 (single-frequency acoustic control) was significantly higher than that of D2, indicating that acoustic intervention itself is beneficial to the fermentation process. However, the characteristic component content of Example 2 (two-stage variable frequency acoustic control) was significantly higher than that of the single high-frequency group (D4), which had the better effect in D4 and D5. Specifically, the total flavonoid and phlorizin content of Example 2 was about 35.7% and 44.4% higher than that of D4, respectively. This difference is not a simple additive result, but rather indicates a significant synergistic effect between "two-stage variable frequency acoustic wave assisted fermentation" (first high frequency to promote bacterial growth, then low frequency to guide metabolism) and "compound enzymatic hydrolysis". This synergistic effect optimizes the metabolic environment of microorganisms at different stages of fermentation, thereby achieving more efficient biotransformation and enrichment of target components.

[0047] It is evident that this invention is not a simple combination of conventional techniques. Data demonstrates that the combination of "compound enzymatic hydrolysis" and "two-stage variable frequency acoustic wave assisted fermentation" significantly improves the efficiency of targeted enrichment of characteristic components.

[0048] Experimental Example 3: To verify the unique role of malt extract in the system of this invention, the enzyme samples prepared in comparative examples D3, D6 and Example 2 (S2) were used as the sole starter and applied to the preparation of standard bread dough. Their effects on the static components and dynamic fermentation activity of the enzyme were compared. The results are shown in Table 3.

[0049] Table 3: Effects of malt extract on enzyme characteristic components and fermentation performance Note: Fermentation time is the length of time it takes for the dough to expand to twice its original size.

[0050] Table 3 shows that the fermentation-promoting effect of malt extract is significant and unique. Even compared with corn extract, which provides similar fermentable sugars, its effect on improving fermentation efficiency (29.2% reduction in time) and bread quality (9.8% increase in specific volume) remains outstanding. This demonstrates that malt extract is not a conventional carbon source in this system, and its specific components synergistically interact with the fermentation system, thereby directionally activating microbial metabolism.

[0051] Experiment Example 4: Furthermore, the above-mentioned enzymes were applied to bread preparation. In order to illustrate the preparation effect, this experiment comprehensively evaluated the application value of the enzyme product (taking S2 as an example) obtained by the complete process of this invention in a real bread making scenario, including independent fermentation ability, synergistic effect and functional component transferability.

[0052] The experimental group design is as follows: Group A (100% Enzyme): Enzyme powder S2 from Example 2 was used to completely replace commercial yeast as a starter culture.

[0053] Group B (50% mixture): Use S2 mixed with commercial yeast at a 1:1 mass ratio as a starter culture.

[0054] Group B1 (30% mixture): Use S2 mixed with commercial yeast at a mass ratio of 3:7 as a starter culture.

[0055] Group C (100% commercial yeast): Commercial yeast was used as the starter culture only (conventional control group).

[0056] The fermentation performance and overall quality of the bread prepared in each group are compared in Table 4: Table 4: Effects of different leavening agents on bread fermentation performance and quality Note: Fermentation time is the length of time it takes for the dough to expand to twice its original size.

[0057] Based on the comprehensive test results in Table 4, the following conclusions can be drawn: The results of Group A indicate that the compound enzyme prepared using the method of this application can independently complete the entire fermentation process of bread, successfully producing high-quality bread with a specific volume of 4.5 mL / g. Simultaneously, it endows the bread with superior anti-aging properties (reduced hardness increase rate) and a unique sensory flavor, proving its basic function as a natural leavening agent. The results of Group B (50% mixture) and Group B1 (30% mixture) together indicate a significant synergistic effect between the enzyme of this invention and commercial yeast. According to the results in Table 4, the combination of the enzyme prepared using the method of this application and commercial yeast can effectively shorten fermentation time and improve bread quality, with the most prominent effect when the mixing ratio is 1:1 (i.e., 50% replacement). Specifically, compared to Group C, which uses 100% commercial yeast, the optimal Group B has a shorter fermentation time, a larger bread specific volume, and the most significant improvement in anti-aging performance (hardness increase rate). The effect of Group B1 is better than Group C but slightly inferior to Group B, indicating that the synergistic effect exists within a certain proportion range and is not a simple linear relationship. Furthermore, flavonoids were detected in the bread crumbs of groups A, B, and B1, but not in group C. This demonstrates that the method described in this application enables the functional plant components in the enzymes to undergo the bread baking process and be successfully transferred to the final food product. This achieves a value upgrade from a "fermenting agent" to a "functional fermenting agent," providing raw material support for the development of baked goods with additional health benefits.

[0058] The above embodiments are merely examples of several implementations of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the present invention.

Claims

1. A method for preparing an edible compound enzyme, characterized in that, Includes the following steps: S1. Compound enzymatic hydrolysis pretreatment: Mix fresh Moringa leaves and fresh Litsea cubeba leaves, crush them, add honey and small molecule water, mix evenly, and then add a compound enzyme solution containing cellulase and pectinase for enzymatic hydrolysis. S2. Two-stage acoustic-assisted fermentation: A compound fermentation agent is inoculated into the material treated in S1 and fermentation is carried out in two stages: A. First stage: Fermentation is carried out with lactic acid bacteria as the main component under the assisted fermentation of sound waves at a frequency of 7500-8500Hz; B. Second stage: Fermentation is carried out with yeast as the main component under the assisted fermentation of sound waves at a frequency of 1000-2000Hz; S3. Activity Stabilization: After fermentation, the fermentation broth is sterilized and concentrated at low temperature to obtain the edible compound enzyme. In the initial stage of S1 or S2, a fermentation promoter accounting for 5%-15% of the total mass of the raw materials is added, and the fermentation promoter is malt extract.

2. The method according to claim 1, characterized in that, In the compound enzymatic hydrolysis pretreatment, the compound enzyme also includes a protease, the enzymatic hydrolysis temperature is 45-55℃, and the enzymatic hydrolysis time is 1-3 hours.

3. The method according to claim 2, characterized in that, In the compound enzymatic hydrolysis pretreatment, the amount of cellulase added is 0.5%-2% of the total mass of the raw materials, the amount of pectinase added is 0.2%-1% of the total mass of the raw materials, and the amount of protease added is 0.1%-0.5% of the total mass of the raw materials.

4. The method according to claim 1, characterized in that, The lactic acid bacteria inoculated in the first stage is Lactobacillus plantarum, and the inoculation amount is 1%-5% (v / v) of the total volume of the fermentation substrate; the yeast inoculated in the second stage is Saccharomyces cerevisiae, and the inoculation amount is 0.5%-3% (v / v) of the total volume of the fermentation substrate.

5. The method according to claim 1, characterized in that, The first stage of fermentation lasts for 24-36 hours, and the second stage of fermentation continues until the pH of the fermentation broth stabilizes at 3.8-4.

2.

6. The method according to claim 1, characterized in that, The sterilization process involves filtration using a 0.22μm microporous membrane.

7. An edible compound enzyme prepared by any one of claims 1 to 6.

8. A method for preparing bread, characterized in that, The edible compound enzyme as described in claim 7 is used in the preparation process.

9. The method according to claim 8, characterized in that, The edible compound enzyme is used as a leavening agent in the bread fermentation process.

10. The method according to claim 8, characterized in that, The edible compound enzyme and commercial yeast are used together as leavening agents in the bread fermentation process, and the mass ratio of the edible compound enzyme to the commercial yeast is 1:1 to 1:2.

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