Bamboo powder fermented feed rich in amino acid and preparation method thereof

Abstract

The invention relates to the field of microbial fermented feed, in particular to bamboo powder fermented feed rich in amino acid and a preparation method thereof. According to the preparation method of the bamboo powder fermented feed provided by the invention, bamboo is used as a base material for fermentation and enzymolysis to prepare amino acid, and then some conventional raw material components of the feed are added to prepare the bamboo powder fermented feed rich in amino acid. The digestibility and the nutrition utilization rate of animals on the feed are higher, the oxidation resistance is stronger, and the palatability is better.

Description

Technical Field

[0001] The invention relates to the field of microbial fermented feed, and in particular to a bamboo powder fermented feed rich in amino acids and a preparation method thereof. Background Art

[0002] As a fast-growing and widely distributed plant resource, bamboo is widely planted and used around the world. Traditionally, bamboo is mainly used in construction, furniture, papermaking and other fields, and bamboo is often regarded as waste and not fully utilized. However, these residues are rich in cellulose, hemicellulose and various trace elements, and have potential feed value.

[0003] In recent years, with the rapid development of animal husbandry, the demand for feed has increased. Traditional feed resources such as grain and forage are facing limitations in land and water resources, and the cost is rising. Therefore, the development of new, low-cost and efficient feed resources has become the key to the sustainable development of animal husbandry. Using bamboo to prepare feed can not only reduce feed costs, but also effectively alleviate the problem of food shortage, while reducing environmental pollution and realizing the recycling of resources.

[0004] In the process of feed preparation, the content of amino acids is one of the important indicators to measure the quality of feed. Amino acids are essential nutrients for animal growth and development, and have an important impact on the growth rate, meat quality, immunity and other aspects of animals. Therefore, how to convert bamboo into amino acid-rich feed through chemical synthesis methods has become the main research goal of the present invention. Summary of the invention

[0005] In view of the above existing technical problems, the present invention aims to provide a bamboo powder fermented feed rich in amino acids and a preparation method thereof, wherein the preparation method uses bamboo as a base material to ferment and enzymatically hydrolyze the bamboo to prepare amino acids, and then adds some conventional feed raw material components to prepare a bamboo powder fermented feed rich in amino acids. The animal has a higher digestibility and nutritional utilization rate of the feed, stronger antioxidant performance, and better palatability.

[0006] The invention discloses a method for preparing a bamboo powder fermented feed rich in amino acids, comprising the following preparation steps: S1 Bamboo pretreatment: The bamboo is screened, cleaned, dried, crushed, and then dried again to obtain bamboo powder; S2 enzymatic hydrolysis treatment: adding the bamboo powder obtained in step S1 to an enzymatic hydrolysis reactor, adding a composite enzyme, and controlling the reaction temperature, pH value and time of the enzymatic hydrolysis reaction to enzymatically hydrolyze the bamboo powder to obtain a small molecule carbohydrate enzymatic hydrolysate; S3 fermentation treatment: the small molecule sugar hydrolysate obtained in step S2 is fed into a fermentation tank, phosphate buffer is added, soybean meal, amino acid fermentation bacteria, vitamins and surfactants are added, and then the fermentation temperature, pH and oxygen content of the fermentation tank are set for fermentation treatment, and fermentation liquid is obtained after the fermentation reaction is completed; S4 separation and purification: the fermentation broth obtained in step S3 is centrifuged, filtered, concentrated and crystallized to separate the amino acids from other impurities in the fermentation broth, and purified to obtain a high-quality amino acid product; S5 feed preparation: add the amino acid product purified in step S4, corn flour, wheat bran, fish meal, sodium chloride and distilled water into a mixer and mix them. After the mixing is complete, heat up and granulate them, and then dry them to obtain bamboo powder fermented feed rich in amino acids.

[0007] Preferably, in the bamboo pretreatment step S1, the particle size of the bamboo powder is 20-50 μm.

[0008] Preferably, in the S2 enzymatic hydrolysis step, the reaction temperature of the enzymatic hydrolysis reaction is 40-60° C., the pH value is 4.5-5.5, and the enzymatic hydrolysis time is 24-48 hours.

[0009] Preferably, in the S2 enzymatic treatment step, the composite enzyme is a combination of ligninase, cellulase and hemicellulase; wherein the mass ratio of ligninase, cellulase and hemicellulase is 1:(1-2):(0.5-1).

[0010] Preferably, in the S3 fermentation treatment step, the amino acid fermentation bacteria is a genetically engineered strain of Corynebacterium glutamicum in which the GlnR gene is overexpressed.

[0011] Preferably, in the S3 fermentation treatment step, the mass ratio of the small molecule carbohydrate hydrolysate to the soybean meal is 1:(0.5-1.5); the mass ratio of the small molecule carbohydrate hydrolysate to the amino acid fermentation bacteria is 1:(2-5).

[0012] Preferably, in the S3 fermentation treatment step, the surfactant is Tween 60.

[0013] Preferably, in the S3 fermentation treatment step, the fermentation pH value of the fermentation tank is 6.5-7.5; the fermentation oxygen content is 20-30%.

[0014] Preferably, in the preparation of S5 feed, the weight portion of the amino acid product is 3 to 5 parts, the weight portion of corn flour is 10 to 13 parts, the weight portion of wheat bran is 10 to 12 parts, the weight portion of fish meal is 1 to 2 parts, and the weight portion of sodium chloride is 0.1 to 0.3 parts.

[0015] A method for preparing a bamboo powder fermented feed rich in amino acids, wherein the bamboo powder fermented feed rich in amino acids is prepared by any of the above methods for preparing a bamboo powder fermented feed rich in amino acids.

[0016] Compared with the prior art, the present invention has the following beneficial effects: The invention provides a bamboo powder fermented feed rich in amino acids and a preparation method thereof. Bamboo is first used as a base material to ferment and enzymolyze to prepare amino acids, and then some conventional feed raw material components are added to prepare the bamboo powder fermented feed rich in amino acids.

[0017] The raw materials used in the method provided by the present invention are widely available. Bamboo, as a rapidly growing and widely distributed plant resource, has a huge amount of residues that are easy to collect. Using these residues to prepare feed can not only reduce feed costs, but also effectively alleviate the problem of food shortages. Bamboo is decomposed into small molecular sugars through enzymatic treatment, and then fermented to synthesize amino acids. The feed prepared by this method has a high amino acid content and can meet the needs of animal growth and development. The method of the present invention realizes the effective utilization of waste bamboo or bamboo materials and reduces environmental pollution. At the same time, the heat and waste gas generated during the enzymatic treatment and fermentation treatment can be recycled, reducing energy consumption. By optimizing the formula design and preparation process, the prepared feed not only has a high amino acid content, but also has balanced nutrition, good taste, and is easy to digest and absorb. This helps to increase the growth rate and immunity of animals and improve meat quality and flavor.

[0018] Surfactants are added during the fermentation process to improve the surface tension of the fermentation liquid, which is more conducive to the growth of the flora and the release of metabolites, promotes the fermentation process, synthesizes amino acid products faster, shortens the feed preparation cycle, improves efficiency and reduces costs. DETAILED DESCRIPTION

[0019] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0020] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0021] Embodiment 1: A method for preparing a bamboo powder fermented feed rich in amino acids, comprising the following steps: S1 Bamboo pretreatment: The bamboo is screened, cleaned, dried, crushed, and dried again to obtain bamboo powder with a particle size of 50 μm.

[0022] S2 enzymatic hydrolysis treatment: adding the bamboo powder obtained in step S1 into an enzymatic hydrolysis reactor, adding a composite enzyme, controlling the reaction temperature of the enzymatic hydrolysis reaction to be 40°C, the pH value to be 4.5, and the enzymatic hydrolysis time to be 24 hours, and enzymatically hydrolyzing the bamboo powder to obtain a small molecule carbohydrate hydrolysate; wherein the composite enzyme is a composition composed of ligninase, cellulase and hemicellulase, and the mass ratio of ligninase, cellulase and hemicellulase is 1:1:0.5.

[0023] S3 fermentation treatment: the small molecule sugar hydrolysate obtained in step S2 is sent to a fermentation tank, and then a phosphate buffer is added, and then soybean meal, amino acid fermentation bacteria, vitamins and surfactant Tween 60 are added, and then the fermentation temperature of the fermentation tank is set to 20°C, the pH value of the fermentation liquid is 6.5, and the oxygen content of the fermentation liquid is 20% for fermentation treatment, and a fermentation liquid is obtained after the fermentation reaction is completed; wherein the amino acid fermentation bacteria is a genetically engineered strain of Corynebacterium glutamicum inlaid with overexpressed GlnR gene; the mass ratio of small molecule sugar hydrolysate to soybean meal is 1:0.5; the mass ratio of small molecule sugar hydrolysate to amino acid fermentation bacteria is 1:2; the mass ratio of small molecule sugar hydrolysate to vitamins is 1:0.1; the mass ratio of the small molecule sugar hydrolysate to surfactant is 1:0.3.

[0024] S4 separation and purification: The fermentation broth obtained in step S3 is subjected to centrifugation, filtration, concentration and crystallization steps to separate the amino acids in the fermentation broth from other impurities, and purify the fermentation broth to obtain a high-quality amino acid product.

[0025] S5 feed preparation: add 3 parts by weight of the amino acid product purified in step S4, 10 parts by weight of corn flour, 10 parts by weight of wheat bran, 1 part by weight of fish meal, and 0.1 parts by weight of sodium chloride into a mixer, then add 20 parts of distilled water to mix, and after the mixing is complete, add the mixture into an extruder for extrusion and granulation, the extrusion temperature is 170°C, the extrusion pressure is 5MPa, the extrusion granulation time is 3h, and after the extrusion and granulation are completed, dry the mixture to obtain a bamboo powder fermented feed rich in amino acids.

[0026] Embodiment 2: A method for preparing a bamboo powder fermented feed rich in amino acids, comprising the following steps: S1 Bamboo pretreatment: The bamboo is screened, cleaned, dried, crushed, and then dried again to obtain bamboo powder with a particle size of 40 μm.

[0027] S2 enzymatic hydrolysis treatment: adding the bamboo powder obtained in step S1 into an enzymatic hydrolysis reactor, adding a composite enzyme, controlling the reaction temperature of the enzymatic hydrolysis reaction to be 50°C, the pH value to be 5, and the enzymatic hydrolysis time to be 35 hours, and enzymatically hydrolyzing the bamboo powder to obtain a small molecule carbohydrate hydrolysate; wherein the composite enzyme is a composition composed of ligninase, cellulase and hemicellulase, and the mass ratio of ligninase, cellulase and hemicellulase is 1:1:0.5.

[0028] S3 fermentation treatment: the small molecule sugar hydrolysate obtained in step S2 is sent to a fermentation tank, and then a phosphate buffer is added, and then soybean meal, amino acid fermentation bacteria, vitamins and surfactant Tween 60 are added, and then the fermentation temperature of the fermentation tank is set to 25°C, the pH value of the fermentation liquid is 7, and the oxygen content of the fermentation liquid is 25% for fermentation treatment, and a fermentation liquid is obtained after the fermentation reaction is completed; wherein the amino acid fermentation bacteria is a genetically engineered strain of Corynebacterium glutamicum inlaid with overexpressed GlnR gene; the mass ratio of small molecule sugar hydrolysate to soybean meal is 1:0.5; the mass ratio of small molecule sugar hydrolysate to amino acid fermentation bacteria is 1:2; the mass ratio of small molecule sugar hydrolysate to vitamins is 1:0.1; the mass ratio of the small molecule sugar hydrolysate to surfactant is 1:0.3.

[0029] S4 separation and purification: The fermentation broth obtained in step S3 is subjected to centrifugation, filtration, concentration and crystallization steps to separate the amino acids in the fermentation broth from other impurities, and purify the fermentation broth to obtain a high-quality amino acid product.

[0030] S5 feed preparation: add 3 parts by weight of the amino acid product purified in step S4, 10 parts by weight of corn flour, 10 parts by weight of wheat bran, 1 part by weight of fish meal, and 0.1 parts by weight of sodium chloride into a mixer, then add 20 parts of distilled water to mix, and after the mixing is complete, add the mixture into an extruder for extrusion and granulation, the extrusion temperature is 170°C, the extrusion pressure is 5MPa, the extrusion granulation time is 3h, and after the extrusion and granulation are completed, dry the mixture to obtain a bamboo powder fermented feed rich in amino acids.

[0031] Embodiment 3: A method for preparing a bamboo powder fermented feed rich in amino acids, comprising the following steps: S1 Bamboo pretreatment: The bamboo is screened, cleaned, dried, crushed, and then dried again to obtain bamboo powder with a particle size of 30 μm.

[0032] S2 enzymatic hydrolysis treatment: adding the bamboo powder obtained in step S1 into an enzymatic hydrolysis reactor, adding a composite enzyme, controlling the reaction temperature of the enzymatic hydrolysis reaction to be 60°C, the pH value to be 5.5, and the enzymatic hydrolysis time to be 48 hours, and enzymatically hydrolyzing the bamboo powder to obtain a small molecule carbohydrate hydrolysate; wherein the composite enzyme is a composition composed of ligninase, cellulase and hemicellulase, and the mass ratio of ligninase, cellulase and hemicellulase is 1:1:0.5.

[0033] S3 fermentation treatment: the small molecule sugar hydrolysate obtained in step S2 is sent to a fermentation tank, and then a phosphate buffer is added, and then soybean meal, amino acid fermentation bacteria, vitamins and surfactant Tween 60 are added, and then the fermentation temperature of the fermentation tank is set to 30°C, the pH value of the fermentation liquid is 7.5, and the oxygen content of the fermentation liquid is 30% for fermentation treatment, and a fermentation liquid is obtained after the fermentation reaction is completed; wherein the amino acid fermentation bacteria is a genetically engineered strain of Corynebacterium glutamicum inlaid with overexpressed GlnR gene; the mass ratio of small molecule sugar hydrolysate to soybean meal is 1:0.5; the mass ratio of small molecule sugar hydrolysate to amino acid fermentation bacteria is 1:2; the mass ratio of small molecule sugar hydrolysate to vitamins is 1:0.1; the mass ratio of the small molecule sugar hydrolysate to surfactant is 1:0.3.

[0034] S4 separation and purification: The fermentation broth obtained in step S3 is subjected to centrifugation, filtration, concentration and crystallization steps to separate the amino acids in the fermentation broth from other impurities, and purify the fermentation broth to obtain a high-quality amino acid product.

[0035] S5 feed preparation: add 3 parts by weight of the amino acid product purified in step S4, 10 parts by weight of corn flour, 10 parts by weight of wheat bran, 1 part by weight of fish meal, and 0.1 parts by weight of sodium chloride into a mixer, then add 20 parts of distilled water to mix, and after the mixing is complete, add the mixture into an extruder for extrusion and granulation, the extrusion temperature is 170°C, the extrusion pressure is 5MPa, the extrusion granulation time is 3h, and after the extrusion and granulation are completed, dry the mixture to obtain a bamboo powder fermented feed rich in amino acids.

[0036] Example 4: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as those in Example 2, except that the mass ratio of ligninase, cellulase and hemicellulase in the S2 enzymatic hydrolysis treatment is changed to 1:1.2:0.6.

[0037] Example 5: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as those in Example 2, except that the mass ratio of ligninase, cellulase and hemicellulase in the S2 enzymatic hydrolysis treatment is changed to 1:1.5:0.7.

[0038] Example 6: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as those in Example 2, except that the mass ratio of ligninase, cellulase and hemicellulase in the S2 enzymatic hydrolysis treatment is changed to 1:1.7:0.8.

[0039] Example 7: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as those in Example 2, except that the mass ratio of ligninase, cellulase and hemicellulase in the S2 enzymatic hydrolysis treatment is changed to 1:2:1.

[0040] Example 8: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as Example 6, except that the mass ratio of small molecule carbohydrate hydrolysate to soybean meal in the S3 fermentation treatment is changed to 1:1.0; the mass ratio of small molecule carbohydrate hydrolysate to amino acid fermentation bacteria is changed to 1:3.5.

[0041] Example 9: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as Example 6, except that the mass ratio of small molecule carbohydrate hydrolysate to soybean meal in the S3 fermentation treatment is changed to 1:1.5; the mass ratio of small molecule carbohydrate hydrolysate to amino acid fermentation bacteria is changed to 1:5.

[0042] Example 10: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as Example 8, except that the weight portions of the components in the preparation of S5 feed are changed: 4 parts of amino acid product, 11 parts of corn flour, 11 parts of wheat bran, 1.5 parts of fish meal, and 0.2 parts of sodium chloride.

[0043] Example 11: A method for preparing a bamboo powder fermented feed rich in amino acids has the same steps and conditions as Example 8, except that the weight portions of the components in the preparation of S5 feed are changed: 5 parts of amino acid product, 13 parts of corn flour, 12 parts of wheat bran, 2 parts of fish meal, and 0.3 parts of sodium chloride.

[0044] The various properties of the amino acid-rich bamboo powder fermented feed prepared in Examples 1 to 11 were tested, and the test results are shown in the following table:

[0045] It can be seen from the above table data that the amino acid content of Example 10 and Example 11 is relatively high; the crude protein content has a relatively consistent trend with the embodiments, and a higher amino acid content usually corresponds to a higher crude protein; the crude fat and crude fiber contents do not change much, the crude fat content remains in the range of 7-8%, and the crude fiber content remains in the range of 12-14%; the carbohydrate content of most embodiments is relatively stable, close to 42%.

[0046] The digestibility and utilization of the bamboo powder fermented feeds prepared in Examples 2, 6, 8 and 11 were evaluated: the feeds were fed to experimental animals (100 sheep) and their digestion was monitored; the digestibility of each feed was measured (through feces collection and feed residue analysis), as well as the utilization efficiency of specific nutrients (such as protein, amino acids, fat, etc.). The experimental data are shown in the following table:

[0047] It can be seen from the data in the above table that the digestibility and amino acid utilization of Example 11 are the highest, which indicates that the digestion and absorption capacity of bamboo powder fermented feed has been improved by optimizing enzymatic hydrolysis and fermentation parameters; the utilization rate of amino acids is positively correlated with the utilization rate of crude protein, indicating that the improvement of amino acids is conducive to the utilization of protein.

[0048] The antioxidant activity of the bamboo powder fermented feeds prepared in Examples 2, 6, 8 and 11 was evaluated: the antioxidant capacity of the feeds in different examples was determined using antioxidant activity test methods such as ABTS and DPPH; the half-inhibitory concentration (IC50) of different feeds was calculated through reaction kinetic analysis. The test results are shown in the following table:

[0049] From the above table data, it can be seen that the lower the IC50 value, the stronger the antioxidant activity. Example 11 has the strongest antioxidant capacity, indicating that it may have a better health support effect on animals.

[0050] The palatability of the bamboo powder fermented feeds prepared in Examples 2, 6, 8 and 11 was evaluated by observing the food intake and feeding time of animals (100 sheep) when given different feeds; and by conducting a feed selection preference experiment, the feed attractiveness of different examples was compared. The experimental data are shown in the following table:

[0051] From the above table data, it can be seen that as the amino acid content and the degree of enzymolysis increase, the palatability of the feed also gradually increases, and Example 9 has the strongest palatability, indicating that it has a high appeal. The bamboo powder fermented feed prepared in Example 11 has the best palatability.

[0052] In summary, the bamboo powder fermented feed prepared in Example 11 is rich in amino acids, has higher digestibility and nutrient utilization rate for animals, stronger antioxidant properties, and better palatability.

[0053] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the scope of protection of the invention.

Claims

1. A method for preparing a bamboo powder fermented feed rich in amino acids, characterized in that: The following steps are involved: S1 Bamboo pretreatment: The bamboo is screened, cleaned, dried, crushed, and then dried again to obtain bamboo powder; S2 enzymatic hydrolysis treatment: adding the bamboo powder obtained in step S1 to an enzymatic hydrolysis reactor, adding a composite enzyme, and controlling the reaction temperature, pH value and time of the enzymatic hydrolysis reaction to enzymatically hydrolyze the bamboo powder to obtain a small molecule carbohydrate enzymatic hydrolysate; S3 fermentation treatment: the small molecule sugar hydrolysate obtained in step S2 is fed into a fermentation tank, phosphate buffer is added, soybean meal, amino acid fermentation bacteria, vitamins and surfactants are added, and then the fermentation temperature, pH and oxygen content of the fermentation tank are set for fermentation treatment, and fermentation liquid is obtained after the fermentation reaction is completed; S4 separation and purification: the fermentation broth obtained in step S3 is centrifuged, filtered, concentrated and crystallized to separate the amino acids from other impurities in the fermentation broth, and purified to obtain a high-quality amino acid product; S5 feed preparation: add the amino acid product purified in step S4, corn flour, wheat bran, fish meal, sodium chloride and distilled water into a mixer and mix them. After the mixing is complete, heat up and granulate them, and then dry them to obtain bamboo powder fermented feed rich in amino acids.

2. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the bamboo pretreatment step S1, the particle size of the bamboo powder is 20-50 μm.

3. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the S2 enzymatic hydrolysis step, the reaction temperature of the enzymatic hydrolysis reaction is 40-60° C., the pH value is 4.5-5.5, and the enzymatic hydrolysis time is 24-48 hours.

4. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the S2 enzymatic hydrolysis step, the composite enzyme is a combination of ligninase, cellulase and hemicellulase; wherein the mass ratio of ligninase, cellulase and hemicellulase is 1:(1-2):(0.5-1).

5. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 4, characterized in that: In the S3 fermentation treatment step, the amino acid fermentation bacteria is a genetically engineered strain of Corynebacterium glutamicum in which the GlnR gene is overexpressed.

6. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the S3 fermentation treatment step, the mass ratio of the small molecule sugar hydrolysate to the soybean meal is 1: (0.5-1.5); the mass ratio of the small molecule sugar hydrolysate to the amino acid fermentation bacteria is 1: (2-5).

7. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the S3 fermentation treatment step, the surfactant is Tween 60.

8. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the S3 fermentation treatment step, the fermentation pH value of the fermentation tank is 6.5-7.5; the fermentation oxygen content is 20-30%.

9. The method for preparing a bamboo powder fermented feed rich in amino acids according to claim 1, characterized in that: In the preparation of S5 feed, the weight portion of the amino acid product is 3 to 5 parts, the weight portion of corn flour is 10 to 13 parts, the weight portion of wheat bran is 10 to 12 parts, the weight portion of fish meal is 1 to 2 parts, and the weight portion of sodium chloride is 0.1 to 0.3 parts.

10. The amino acid-rich bamboo powder fermented feed prepared by the method for preparing amino acid-rich bamboo powder fermented feed according to any one of claims 1 to 9.

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