Preparation method of high-activity feed of plant chenopodium based on synergistic fermentation of bacteria and enzymes, fermented plant chenopodium feed and application

Abstract

The application provides a preparation method of high-activity plant goosefoot feed based on synergistic fermentation of bacteria and enzymes, fermented plant goosefoot feed and application, and relates to the technical field of biological fermentation feed preparation. The method comprises the following steps: mixing plant goosefoot, corn flour and cinnamon powder to obtain a fermentation substrate; adding compound probiotics and pectinase to the fermentation substrate and adjusting the water content; the compound probiotics comprise bacillus subtilis, lactobacillus plantarum and saccharomyces cerevisiae; performing solid-state fermentation on the fermentation material, so that the pectinase enzymatically degrades and destroys the cell wall structure of the plant goosefoot, and the compound probiotics ferment and convert the plant goosefoot; and after drying and crushing, the fermented plant goosefoot feed is obtained. The application can improve the release degree of soluble active ingredients in the plant goosefoot.

Description

Technical Field

[0001] This invention relates to the field of bio-fermented feed preparation technology, and in particular to a method for preparing a highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes, the fermented plant-based quinoa feed, and its applications. Background Technology

[0002] Chenopodium album L. is a widely distributed natural herb with high nutritional value and medicinal potential, rich in protein, dietary fiber, polysaccharides, polyphenols, and other bioactive components. Chenopodium album L. exhibits beneficial physiological functions in antioxidation, anti-inflammation, and immune regulation, thus possessing significant development and utilization value in the feed and functional additive fields. However, untreated chenopodium album L. still faces several limitations in practical applications. On the one hand, its cell wall structure is relatively dense, with active ingredients mostly existing in bound forms, resulting in low release rates and poor bioavailability of functional components such as polysaccharides and phenols. On the other hand, chenopodium album L. contains certain amounts of anti-nutritional factors, such as pectin, phytic acid, and tannins, which can affect the digestion and absorption of nutrients by animals, thereby limiting its efficient utilization in feed. Current research mainly focuses on chenopodium album L. extracts or simple processing, lacking research on systematically improving the overall nutritional structure, active components, and functional characteristics of chenopodium album L. using biotechnology.

[0003] Currently, microbial fermentation technology is widely used in feed processing due to its ability to degrade macromolecules, release active ingredients, and improve feed palatability. During microbial metabolism, large protein molecules are degraded, releasing small peptides and free amino acids, while simultaneously altering the metabolic pathways of lipids and carbohydrates, thereby improving feed flavor and structural characteristics. However, existing microbial fermentation technologies still suffer from the following problems: most processes do not effectively pre-treat raw materials, resulting in insufficient cell wall disruption and hindering the effective release of active ingredients; many current fermentation processes rely on single strains or simple combinations of microorganisms, limiting the conversion efficiency of active ingredients; and most existing fermentation processes focus only on increasing the total amount of active substances, failing to effectively regulate structural changes in active ingredients. Furthermore, while enzymatic hydrolysis can degrade plant cell walls or specific substrates, its use alone is easily limited by substrate structure and lacks the synergistic effect of continuous metabolic processes. To address these issues, microbial-enzyme co-fermentation technology has gradually gained attention. This technology combines microbial fermentation with enzymatic hydrolysis to rapidly disrupt plant cell wall structures, while simultaneously further transforming substrates through microbial metabolism, achieving deep modification of nutrients and active substances, thus significantly improving fermentation efficiency. However, currently, the co-fermentation technology of bacteria and enzymes is mainly applied to conventional feed ingredients such as soybean meal and wheat bran, while there is little systematic research on natural herbaceous plants such as quinoa. There is also a lack of in-depth research on the optimized fermentation process and mechanism of action of these plants, which are based on their unique structural characteristics and functional components.

[0004] Therefore, it is necessary to develop a microbial-enzyme co-fermentation technology for quinoa, rationally construct a synergistic system of microorganisms and enzymes, and optimize the cell structure, active substance composition and functional characteristics of quinoa, thereby improving its microecological function and enhancing its nutritional value and functionality in feed. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing a highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes, as well as the fermented plant-based quinoa feed and its application. This method can solve the problems of insufficient release of polysaccharides and phenolic active ingredients due to the dense cell wall structure of plant-based quinoa, lack of continuous metabolic transformation due to simple enzymatic hydrolysis, and insufficient cell wall disruption due to simple fermentation. It achieves synergistic enzymatic destruction of the cell wall structure of plant-based quinoa and fermentation transformation, improves the release of soluble active ingredients in plant-based quinoa, and obtains a fermented plant-based quinoa feed suitable for preparing broiler feed or broiler feed additives.

[0006] To achieve the above objectives, the present invention provides the following solution: This invention provides a method for preparing a highly bioactive plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes, comprising: The plant quinoa, corn flour, and cinnamon powder were mixed to obtain a fermentation substrate with plant quinoa as the main raw material. Compound probiotics and pectinase are added to the fermentation substrate, and the moisture content of the fermentation substrate is adjusted to obtain the material to be fermented; the compound probiotics include Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae; Solid-state fermentation is carried out on the material to be fermented, so that pectinase enzymatically breaks down the cell wall structure of the plant quinoa, and compound probiotics ferment and transform the enzymatically broken plant quinoa to obtain fermentation products. The fermentation products are dried and pulverized to obtain fermented quinoa feed.

[0007] Preferably, the roots of the quinoa plant are removed before it is mixed with corn flour and cinnamon powder, and then it is dried and pulverized.

[0008] Preferably, the quinoa, corn flour, and cinnamon powder are mixed in a mass ratio of 75.5:20:0.5.

[0009] Preferably, the ratio of Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae is 1:1:1.

[0010] Preferably, the amount of compound probiotics added is 1‰ of the fermentation substrate mass.

[0011] Preferably, the amount of pectinase added is 4% based on enzyme activity.

[0012] Preferably, adjusting the water content of the fermentation substrate includes adding distilled water to the fermentation substrate containing compound probiotics and pectinase, wherein the amount of distilled water added is 50% of the mass of the fermentation substrate.

[0013] Preferably, the solid-state fermentation temperature is 26°C to 32°C, and the fermentation time is 24h to 72h.

[0014] Another aspect of the present invention provides a fermented plant quinoa feed, which is prepared by the above-mentioned method for preparing a highly active plant quinoa feed based on synergistic fermentation of bacteria and enzymes.

[0015] In another aspect, the present invention provides the application of the above-mentioned fermented plant quinoa feed in the preparation of broiler feed or broiler feed additives.

[0016] The present invention discloses the following beneficial effects: This invention addresses the issue of insufficient release of active ingredients from quinoa by formulating a fermentation substrate with quinoa as the main raw material, using quinoa, corn flour, and cinnamon powder as the main ingredients. This is achieved by confining the fermentation process to a fermentable and enzymatically hydrolyzable substrate system. By adding a compound probiotic containing Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae, along with pectinase, and adjusting the substrate's moisture content, the pectinase can act on the cell wall structure of quinoa, reducing the restriction of polysaccharide and phenolic substances release by the dense cell wall. Simultaneously, the compound probiotics ferment and transform the enzymatically hydrolyzed quinoa in the same solid-state fermentation system, mitigating the problems of insufficient continuous metabolic transformation from simple enzymatic hydrolysis and insufficient cell wall disruption from simple fermentation. Finally, by solid-state fermentation of the material to be fermented and drying and pulverizing the fermentation products, a fermented quinoa feed suitable for use as animal feed is obtained. Under the exemplary preparation conditions, the polysaccharide content of quinoa increased from 147.36±9.90 mg / g to 200.13±11.38 mg / g after fermentation, and the polyphenol content increased from 12.53±2.16 mg / g to 17.82±2.03 mg / g. This indicates that the above-mentioned bacterial-enzyme synergistic solid-state fermentation process can promote the release of soluble active ingredients in quinoa and transform the quinoa tissue from a relatively dense state to a relatively loose state. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 A flowchart of the method provided in an embodiment of the present invention; Figure 2 This is a comparison chart of polysaccharide and polyphenol content before and after fermentation of quinoa plants provided in an embodiment of the present invention; Figure 3 The images provided in this embodiment of the invention are scanning electron microscope images of quinoa plant before and after fermentation, wherein the upper left and lower left images are of quinoa plant before fermentation, and the upper right and lower right images are of quinoa plant after fermentation. Figure 4 A representative diagram of broiler chicken small intestinal cells provided in an embodiment of the present invention; Figure 5 This is a graph showing the effect of different types of enzymes on the polysaccharide and polyphenol content of fermented quinoa feed provided in the embodiments of the present invention; Figure 6 This is a graph showing the effect of different doses of pectinase on the polysaccharide and polyphenol content of fermented quinoa feed, as provided in an embodiment of the present invention. Figure 7The graph shows the effect of fermentation temperature on the polysaccharide and polyphenol content of fermented quinoa feed, as provided in an embodiment of the present invention. Detailed Implementation

[0019] 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.

[0020] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0021] Figure 1 The method flowchart provided in the embodiments of the present invention is as follows: Figure 1 As shown, this embodiment provides a method for preparing a highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes, comprising: Step 100: Mix quinoa, corn flour and cinnamon powder to obtain a fermentation substrate with quinoa as the main raw material; Step 200: Add compound probiotics and pectinase to the fermentation substrate and adjust the moisture content of the fermentation substrate to obtain the material to be fermented; the compound probiotics include Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae; Step 300: Solid-state fermentation is carried out on the material to be fermented, so that pectinase enzymatically destroys the cell wall structure of the plant quinoa, and compound probiotics ferment and transform the enzymatically destroyed plant quinoa to obtain the fermentation product. Step 400: Dry and pulverize the fermentation product to obtain fermented quinoa feed.

[0022] In step 100, after harvesting the quinoa plant and removing the roots, the plant is dried and pulverized to serve as the main raw material for the fermentation substrate. Quinoa is rich in active ingredients such as protein, dietary fiber, polysaccharides, and polyphenols. However, the cell wall structure of untreated quinoa is relatively dense, and the active ingredients mostly exist in bound form, resulting in a low release rate of polysaccharides and phenols. By mixing quinoa, corn flour, and cinnamon powder to form the fermentation substrate, the problem of insufficient release of active ingredients from quinoa is addressed within a single substrate system. Corn flour provides the available carbon source required for fermentation, while cinnamon powder serves as an auxiliary ingredient in the composition of the fermentation substrate.

[0023] In one specific embodiment, quinoa, corn flour, cinnamon powder, and pectinase are mixed in a mass ratio of 75.5:20:0.5:4. This mass ratio corresponds to a preferred formulation. In actual preparation, the dried quinoa can be pulverized first to ensure sufficient contact between the quinoa and the auxiliary materials (corn flour and cinnamon powder), reducing the impact of uneven distribution of components within the fermentation substrate on subsequent synergistic fermentation by bacteria and enzymes.

[0024] In step 200, a compound probiotic and pectinase are added to the fermentation substrate, and the moisture content of the substrate is adjusted to obtain the material to be fermented. The compound probiotic includes Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae. Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae can form a compound microbial fermentation system in a solid-state fermentation system. Pectinase can act on the pectin-like structural components in the cell walls of quinoa plants, causing enzymatic degradation of the cell wall structure. The compound probiotic and pectinase are added to the fermentation substrate simultaneously, allowing the enzymatic degradation and cell wall disruption process and the microbial fermentation transformation process to occur in the same solid-state fermentation system, which differs from the treatment method of first performing independent enzymatic degradation and then fermenting the enzymatic degradation products.

[0025] In one specific embodiment, the ratio of Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae is 1:1:1, and the amount of compound probiotics added is 1‰ of the fermentation substrate. The amount of pectinase added is 4%. After the compound probiotics and pectinase are added to the fermentation substrate, they are mixed to ensure that the pectinase fully contacts the cell wall of the plant quinoa and that the compound probiotics are evenly distributed in the fermentation substrate. The pectinase was purchased from Beijing Solai Biotechnology Co., Ltd. (Beijing, China), and its specification is 100,000 U / g.

[0026] In one specific embodiment, when adjusting the moisture content of the fermentation substrate, distilled water is added to the fermentation substrate containing compound probiotics and pectinase, with the amount of distilled water being 50% of the substrate mass. After adding distilled water, the fermentation substrate transforms into a material suitable for solid-state fermentation. If the moisture content is too low, pectinase and compound probiotics will have difficulty fully diffusing in the fermentation substrate; if the moisture content is too high, the aeration and material structure stability of the solid-state fermentation system will be easily affected. Therefore, moisture content adjustment is used to balance the conditions for pectinase activity and the fermentation conditions for compound probiotics.

[0027] In step 300, the material to be fermented undergoes solid-state fermentation. During solid-state fermentation, pectinase enzymatically breaks down the cell wall structure of the quinoa plant, gradually loosening the dense quinoa tissue structure. The compound probiotics utilize the enzymatically broken-down quinoa plant and the available nutrients in the fermentation substrate for fermentation transformation, promoting the release of polysaccharides and phenolic active substances from the quinoa plant. Through this process, the problems of insufficient continuous metabolic transformation from simple enzymatic hydrolysis and insufficient cell wall disruption from simple fermentation can be simultaneously improved.

[0028] In one specific embodiment, the solid-state fermentation temperature is 26°C to 32°C, and the fermentation time is 24 hours to 72 hours. In a preferred embodiment, the solid-state fermentation temperature is 28°C, and the fermentation time is 24 hours. After fermentation, a fermentation product is obtained. The plant quinoa tissue structure in this fermentation product is more porous than before fermentation, which is beneficial for the release of active substances and subsequent utilization as feed.

[0029] In step 400, the fermentation product is dried and pulverized to obtain fermented quinoa feed. The drying process is used to reduce the impact of the moisture content of the fermentation product on storage and subsequent use, while the pulverization process is used to obtain fermented quinoa feed with more uniform particle size, enabling it to be used as broiler feed or broiler feed additive.

[0030] Example 1 This embodiment provides a method for preparing fermented quinoa feed.

[0031] The roots of the harvested quinoa plants were removed, dried, and pulverized for later use. A fermentation substrate was prepared by mixing 75.5 parts quinoa, 20 parts corn flour, and 0.5 parts cinnamon powder. A compound probiotic and pectinase were added to the fermentation substrate. The compound probiotic included Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae, with a strain ratio of 1:1:1. The amount of compound probiotic added was 1‰ of the substrate weight, and the amount of pectinase added was 4%. Distilled water was added to the fermentation substrate containing the compound probiotic and pectinase, at a volume of 50% of the substrate weight. The mixture was then used to obtain the material to be fermented. The material was fermented in a solid state at 28℃ for 24 hours. After fermentation, it was dried and pulverized to obtain fermented quinoa feed.

[0032] In the fermented quinoa feed obtained in this embodiment, the polysaccharide and polyphenol contents are increased compared with the quinoa before fermentation, and the quinoa tissue structure changes from a denser state to a looser state.

[0033] Detection of active ingredients before and after fermentation The polysaccharide and polyphenol contents of unfermented and fermented quinoa feed were tested, and the results are as follows: Figure 2 As shown. Figure 2 This is a graph comparing the polysaccharide and polyphenol content of quinoa before and after fermentation. Figure 2In this study, CAL represents unfermented quinoa plant, and FCAL represents fermented quinoa plant feed. The polysaccharide content of unfermented quinoa plant was 147.36±9.90 mg / g, and the polyphenol content was 12.53±2.16 mg / g; the polysaccharide content of fermented quinoa plant feed was 200.13±11.38 mg / g, and the polyphenol content was 17.82±2.03 mg / g. The total polysaccharide content of quinoa plant increased by approximately 35.81% after fermentation. 206 significantly different phenolic compounds were detected during fermentation, of which 79 were significantly upregulated. These results indicate that the synergistic effect of compound probiotics and pectinase in the same solid-state fermentation system can promote the release of soluble polysaccharides and polyphenols from quinoa plant and alter the composition of phenolic substances.

[0034] Microstructure observation before and after fermentation The microstructure of unfermented quinoa and fermented quinoa feed was observed using scanning electron microscopy. During the examination, the unfermented quinoa and fermented quinoa feed were fixed, dried, and sputter-coated with gold, respectively, before being observed under a scanning electron microscope at magnifications of 1000x and 50 μm, respectively. The results are shown below. Figure 3 As shown. Figure 3 These are scanning electron microscope (SEM) images of quinoa plants before and after fermentation. The top left and bottom left images show quinoa plants before fermentation, while the top right and bottom right images show the fermented quinoa feed. Figure 3 It is evident that the surface structure of quinoa plants was relatively compact before fermentation, with irregular particles present. After fermentation, the spatial structure of quinoa plants changed, with increased surface fracturing, looser tissue structure, and an increased number of cavities. These results indicate that the synergistic solid-state fermentation of compound probiotics and pectinase can disrupt the cell walls and tissue structure of quinoa plants, and this structural change provides a physical basis for the release of polysaccharides and polyphenols.

[0035] Animal feeding trials Fermented quinoa feed was used as a basal dietary supplement for broilers, with four addition levels (0 g / kg, 2 g / kg, 4 g / kg, and 8 g / kg) to evaluate its effects on growth performance, blood parameters, and intestinal morphology. Experimental data were calculated and processed using Excel 2021. One-way ANOVA was performed using SAS 9.2 software, and Duncan's test was used for multiple comparisons. Results are expressed as mean and standard error, with P < 0.05 considered statistically significant.

[0036] The effects of feeding fermented quinoa feed on broiler growth performance are shown in Table 1. Table 1 shows the effects of fermented quinoa feed on broiler growth performance. The brooding period was from 1 to 21 days of age. During the brooding period, the fermented quinoa feed supplementation groups had significant effects on the average daily weight gain and feed conversion ratio of broilers. Compared with the 0 g / kg supplementation, 4 g / kg and 8 g / kg of fermented quinoa feed significantly increased the average daily weight gain of broilers, while 8 g / kg significantly decreased the feed conversion ratio. The rearing period was from 22 to 42 days of age. During the rearing period, 2 g / kg and 4 g / kg of feed significantly increased the average daily weight gain of broilers, while 2 g / kg significantly decreased the feed conversion ratio. Throughout the entire 1 to 42 day period, the average daily weight gain of broilers corresponding to 4 g / kg and 8 g / kg supplementation was significantly higher than that of 0 g / kg supplementation.

[0037] Table 1. Effects of fermented quinoa feed on the growth performance of broiler chickens Note: In the same row of data, different subheadings indicate significant differences (P<0.05); the same subheading indicates no significant differences. Subheadings are not repeated in the table for ease of explanation.

[0038] The effects of feeding fermented quinoa feed on broiler blood parameters are shown in Table 2. Table 2 presents the results of blood parameter tests in 42-day-old broilers. On day 42, the serum insulin-like growth factor-1 (IGF-1) level in broilers increased with the amount of fermented quinoa feed added, showing a quadratic curve. The IGF-1 level in the fermented quinoa feed group was significantly higher than that at the 0 g / kg addition level. The serum immunoglobulin A (IGA) level in broilers also increased with the amount of fermented quinoa feed added, showing a quadratic curve. The IGA level in the fermented quinoa feed group was significantly higher than that at the 0 g / kg addition level. Addition levels of 2 g / kg and 4 g / kg significantly reduced the serum interleukin-10 level in broilers.

[0039] Table 2. Effects of fermented quinoa feed on blood parameters in broiler chickens The effects of feeding fermented quinoa feed on broiler intestinal morphology are shown in [reference needed]. Figure 4 And Table 3. Figure 4 This is a representative image of small intestinal cells in 42-day-old broiler chickens, magnified 2×, with a scale bar of 200 μm. Figure 4 It is evident that the intestinal villi of 42-day-old broiler chickens with 0 g / kg addition and fermented quinoa feed addition were well developed and clearly demarcated, with a large number of villi per unit field of view. No structural damage was observed in any group, indicating that fermented quinoa feed additions of 2 g / kg to 8 g / kg did not cause structural damage to the small intestine of broiler chickens.

[0040] Table 3 shows the intestinal morphology results of 42-day-old broiler chickens. On day 42, fermented quinoa feed significantly affected duodenal villus height, crypt depth in the three intestinal segments, and the villus height-to-crypt depth ratio. Compared to the 0 g / kg addition, the fermented quinoa feed group showed a significantly higher villus height-to-crypt depth ratio in all three intestinal segments, and a significantly lower crypt depth in the duodenum and ileum. Duodenal villus heights at 4 g / kg and 8 g / kg additions were significantly higher than those at 0 g / kg addition, while jejunal crypt depths at 8 g / kg addition were significantly lower than those at 0 g / kg addition.

[0041] Table 3. Effects of fermented quinoa feed on intestinal morphology in broiler chickens Comparative Example 1: The Effect of Different Enzymes on Fermentation Efficiency To screen enzyme preparations suitable for solid-state fermentation systems of quinoa, quinoa plants were treated with β-glucanase, cellulase, pectinase, β-mannanase, xylanase, and acidic protease, respectively, and fermented under the same conditions. The polysaccharide and polyphenol contents in the fermented products were measured, and the results are as follows: Figure 5 As shown. Figure 5 This figure shows the effect of different types of enzymes on the polysaccharide and polyphenol content of fermented quinoa feed. Figure 5 It is evident that the β-glucanase treatment resulted in the highest polysaccharide content in the fermented quinoa feed, reaching 160.74 mg / g, significantly higher than the treatments using cellulase, pectinase, β-mannanase, xylanase, and acidic protease. Regarding polyphenol content, the pectinase treatment also yielded the highest polyphenol content, reaching 14.83 mg / g, significantly higher than the treatments using acidic protease, cellulase, xylanase, β-glucanase, and β-mannanase. Considering both polysaccharide and polyphenol content in the fermented quinoa feed, the pectinase treatment group maintained high levels of both polysaccharide and polyphenol release. Therefore, pectinase was selected as the enzyme preparation for the solid-state fermentation system of quinoa.

[0042] Comparative Example 2: The effect of pectinase addition on fermentation efficiency Fermentation experiments were conducted using pectinase at three dosages: 2000U, 3000U, and 4000U. The polysaccharide and polyphenol contents in the fermented quinoa feed were measured, and the results are as follows: Figure 6 As shown. Figure 6 The graph shows the effect of different doses of pectinase on the polysaccharide and polyphenol content of fermented quinoa feed. Figure 6It is evident that the polysaccharide and polyphenol contents in the fermented quinoa feed increased with increasing pectinase addition. When the pectinase addition was 4000U, the polysaccharide content in the fermented quinoa feed was significantly higher than in other treatment groups; similarly, when the pectinase addition was 3000U and 4000U, the polyphenol content in the fermented quinoa feed was significantly higher than in other treatment groups. Considering both polysaccharide and polyphenol contents, 4000U was selected as the optimal pectinase addition level.

[0043] Comparative Example 3: The Effect of Fermentation Temperature on Fermentation Efficiency Under the same ingredients, compound probiotics, and pectinase addition conditions, four fermentation temperatures (26℃, 28℃, 30℃, and 32℃) were set, and the polysaccharide and polyphenol contents in the fermented product were measured. The results are as follows: Figure 7 As shown. Figure 7 This graph shows the effect of fermentation temperature on the polysaccharide and polyphenol content of fermented quinoa feed. Figure 7 It is evident that the order of polysaccharide and polyphenol content in fermented quinoa feed is 30℃ < 32℃ < 26℃ < 28℃. There was no significant difference in polyphenol content between the 26℃ and 28℃ groups, but both were significantly higher than those in the 30℃ and 32℃ groups. Considering both polysaccharide and polyphenol content, 28℃ is the optimal fermentation temperature.

[0044] As demonstrated in the above examples and comparative examples, after mixing quinoa, corn flour, and cinnamon powder to form a fermentation substrate with quinoa as the main raw material, simultaneously adding a compound probiotic containing Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae, as well as pectinase, and conducting solid-state fermentation after adjusting the moisture content, allows the pectinase to enzymatically break down the cell wall structure of quinoa, and enables the compound probiotics to ferment and transform the enzymatically broken-down quinoa. This process can increase the release of soluble polysaccharides and polyphenols from quinoa, and the fermentation product, after drying and pulverizing, forms a fermented quinoa feed suitable for broiler feed or broiler feed additives.

[0045] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0046] Specific examples have been used to illustrate the principles and implementation methods of this invention. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this invention. Furthermore, those skilled in the art will recognize that, based on the ideas of this invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of this invention.

Claims

1. A method for preparing a highly active plant-based feed of quinoa based on synergistic fermentation of bacteria and enzymes, characterized in that, include: The plant quinoa, corn flour, and cinnamon powder are mixed to obtain a fermentation substrate with the plant quinoa as the main raw material. A compound probiotic and pectinase are added to the fermentation substrate, and the moisture content of the fermentation substrate is adjusted to obtain the material to be fermented; the compound probiotic includes Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae; The material to be fermented is subjected to solid-state fermentation, in which the pectinase enzymatically destroys the cell wall structure of the plant quinoa, and the compound probiotics ferment and transform the enzymatically destroyed plant quinoa to obtain the fermentation product. The fermentation product is dried and pulverized to obtain fermented quinoa feed.

2. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, Before being mixed with the corn flour and cinnamon powder, the roots of the quinoa plant are removed, and it is then dried and pulverized.

3. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, The quinoa plant, the corn flour, and the cinnamon powder are mixed in a mass ratio of 75.5:20:0.

5.

4. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, The ratio of Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae is 1:1:

1.

5. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, The amount of the compound probiotic added is 1‰ of the fermentation substrate.

6. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, The amount of pectinase added is 4% based on enzyme activity.

7. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, Adjusting the water content of the fermentation substrate includes adding distilled water to the fermentation substrate containing the compound probiotics and the pectinase, wherein the amount of distilled water added is 50% of the mass of the fermentation substrate.

8. The method for preparing highly active plant-based quinoa feed based on synergistic fermentation of bacteria and enzymes according to claim 1, characterized in that, The solid-state fermentation temperature is 26℃ to 32℃, and the fermentation time is 24h to 72h.

9. A fermented plant-based quinoa feed, characterized in that, The feed is prepared by the method for preparing plant-based high-activity feed based on bacterial-enzyme co-fermentation as described in any one of claims 1 to 8.

10. The use of the fermented quinoa feed according to claim 9 in the preparation of broiler feed or broiler feed additive.

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