Compound glycosyl biological feed leavening agent and preparation method and application thereof
阅读说明：本技术 一种复合糖基生物饲料发酵剂及其制备方法与应用 (Compound glycosyl biological feed leavening agent and preparation method and application thereof ) 是由 左瑞华 蒋平 何燕飞 王敬利 孙桃桃 徐光沛 于 2021-09-26 设计创作，主要内容包括：本发明公开了一种复合糖基生物饲料发酵剂,其原料包括：糖基质、益生菌和酶制剂；且三者的重量份配比为20:2:1；其中,糖基质包括下述质量分数的成分：葡萄糖30-50％、果糖10-20％、乳糖5-15％、麦芽糖15-25％和蔗糖10-20％；益生菌包括下述质量分数的成分：枯草芽孢杆菌5-15％、地衣芽孢杆菌5-15％、植物乳杆菌20-30％、发酵乳杆菌15-25％、乳酸链球菌1-10％、啤酒酵母菌20-30％、双歧杆菌5-10％；酶制剂包括下述质量分数的成分：纤维素酶45-65％和单宁酶40-55％。(The invention discloses a composite glycosyl biological feed leaven, which comprises the following raw materials: sugar substrates, probiotics and enzyme preparations; the weight ratio of the three components is 20:2: 1; wherein the sugar matrix comprises the following components in percentage by mass: 30-50% of glucose, 10-20% of fructose, 5-15% of lactose, 15-25% of maltose and 10-20% of sucrose; the probiotics comprise the following components in percentage by mass: 5-15% of bacillus subtilis, 5-15% of bacillus licheniformis, 20-30% of lactobacillus plantarum, 15-25% of lactobacillus fermentum, 1-10% of streptococcus lactis, 20-30% of saccharomyces cerevisiae and 5-10% of bifidobacterium; the enzyme preparation comprises the following components in percentage by mass: 45-65% of cellulase and 40-55% of tannase.)
1. The composite glycosyl biological feed leaven is characterized by comprising the following raw materials: sugar substrates, probiotics and enzyme preparations; the weight ratio of the three components is 10-25:0.5-4: 0.1-2;
wherein the sugar matrix comprises the following components in percentage by mass: 30-50% of glucose, 10-20% of fructose, 5-15% of lactose, 15-25% of maltose and 10-20% of sucrose;
the probiotics comprise the following components in percentage by mass: 5-15% of bacillus subtilis, 5-15% of bacillus licheniformis, 20-30% of lactobacillus plantarum, 15-25% of lactobacillus fermentum, 1-10% of streptococcus lactis, 20-30% of saccharomyces cerevisiae and 5-10% of bifidobacterium;
the enzyme preparation comprises the following components in percentage by mass: 45-65% of cellulase and 40-55% of tannase.
2. The complex glycosyl biological feed starter as claimed in claim 1, wherein the sugar substrate comprises the following components in percentage by mass: 45% of glucose, 15% of fructose, 10% of lactose, 19% of maltose and 11% of sucrose.
3. The complex glycosyl biological feed starter as claimed in claim 2, wherein the probiotic comprises the following components in percentage by mass: 8% of bacillus subtilis, 12% of bacillus licheniformis, 23% of lactobacillus plantarum, 17% of lactobacillus fermentum, 5% of streptococcus lactis, 28% of saccharomyces cerevisiae and 7% of bifidobacterium.
4. The complex glycosyl biological feed starter as claimed in claim 3, wherein the enzyme preparation comprises the following components in percentage by mass: 58% of cellulase and 42% of tannase.
5. The method for preparing the complex glycosyl biological feed leaven of any one of claims 1 to 4, which is characterized by comprising the following steps:
1) weighing: weighing the raw materials according to the weight part ratio, and storing the raw materials separately for later use;
2) preparing a liquid sugar matrix: adding sugar substrate and water into a liquid fermentation tank to prepare liquid sugar substrate;
3) liquid state fermentation: adding probiotics into the liquid sugar matrix, and performing liquid fermentation at 25-35 ℃ for 3-5 days to obtain fermentation liquor;
4) preparing a fermentation preparation: adding an enzyme preparation into the fermentation liquor, and uniformly mixing to obtain the composite glycosyl biological feed leavening agent.
6. The method for preparing the composite sugar-based biological feed starter according to claim 5, wherein the mass ratio of the sugar substrate to the water in the step 2) is 1: 5.
7. The use of the complex glycosyl biological feed leaven prepared by the preparation method of claim 5 or 6 in the preparation of biological feed.
8. A method for preparing biological feed is characterized by comprising the following steps:
pulverizing and softening silage raw materials, adding the compound glycosyl biological feed starter, and hermetically storing at 20-35 ℃ to obtain the biological feed.
9. The method for preparing the biological feed according to claim 8, wherein the mass ratio of the silage to the compound glycosyl biological feed starter is 100: 1; the time for sealed storage was 15 d.
10. The method of claim 8, wherein the process of pulverizing and softening the silage raw material comprises: cutting the raw materials into 2-3cm segments, bundling the segments, sealing with sealed bags, stacking, coating film, and fermenting at 20-35 deg.C for 7 days or longer.
Technical Field
The invention relates to the technical field of biological feed leavening agents, in particular to a compound glycosyl biological feed leavening agent and a preparation method and application thereof.
Background
The biological feed refers to a general term of feed products developed by biological engineering techniques such as fermentation engineering, enzyme engineering, protein engineering and genetic engineering using feed raw materials and additives allowed by relevant national regulations. In actual production, after the plant raw materials are collected, microorganisms such as lactic acid bacteria, saccharomycetes and bacillus subtilis are mostly used for anaerobic fermentation to obtain the biological fermentation feed, and the feed has the advantages of good palatability, rich probiotic content, easiness in absorption and the like, and is always the preferred mode for storing the green feed for the ruminants.
However, when the conventional fermentation preparation is used for silage fermentation of feed, fermentation flora, sugar substrate addition and the like have some problems, so that the number of dominant probiotic flora is unstable, the substrate mixing is not uniform, and the fermentation efficiency of the biological fermentation feed is unstable.
Therefore, it is an urgent need to solve the problem of providing a fermentation preparation that can maintain a stable number of dominant bacteria and improve the fermentation efficiency of fermented feeds.
Disclosure of Invention
In view of the above, the invention provides a composite glycosyl biological feed starter, a preparation method and application thereof, the starter can highlight the number of dominant floras such as lactic acid bacteria, and the like, the preparation process is simple, and the fermentation efficiency and the fermentation quality of biological feed are ensured.
In order to achieve the purpose, the invention adopts the following technical scheme:
a compound glycosyl biological feed leaven comprises the following raw materials: sugar substrates, probiotics and enzyme preparations; the weight ratio of the three components is 10-25:0.5-4: 0.1-2; the sugar substrate provides nutrition for the rapid propagation of probiotics, and the enzyme preparation mainly decomposes cellulose and tannin;
wherein the sugar matrix comprises the following components in percentage by mass: 30-50% of glucose, 10-20% of fructose, 5-15% of lactose, 15-25% of maltose and 10-20% of sucrose;
the probiotics comprise the following components in percentage by mass: 5-15% of bacillus subtilis, 5-15% of bacillus licheniformis, 20-30% of lactobacillus plantarum, 15-25% of lactobacillus fermentum, 1-10% of streptococcus lactis, 20-30% of saccharomyces cerevisiae and 5-10% of bifidobacterium;
the enzyme preparation comprises the following components in percentage by mass: 45-65% of cellulase and 40-55% of tannase.
As a preferable technical solution of the above technical solution, the sugar base comprises the following components in mass fraction: 45% of glucose, 15% of fructose, 10% of lactose, 19% of maltose and 11% of sucrose.
As a preferable technical scheme of the above technical scheme, the probiotic comprises the following components in percentage by mass: 8% of bacillus subtilis, 12% of bacillus licheniformis, 23% of lactobacillus plantarum, 17% of lactobacillus fermentum, 5% of streptococcus lactis, 28% of saccharomyces cerevisiae and 7% of bifidobacterium.
As a preferable technical scheme of the technical scheme, the enzyme preparation comprises the following components in percentage by mass: 58% of cellulase and 42% of tannase.
The invention also claims a preparation method of the compound glycosyl biological feed leaven, which is the same as the above subject matter, and comprises the following processes:
1) weighing: weighing the raw materials according to the weight part ratio, and storing the raw materials separately for later use;
2) preparing a liquid sugar matrix: adding sugar substrate and water into a liquid fermentation tank to prepare liquid sugar substrate;
3) liquid state fermentation: adding probiotics into the liquid sugar matrix, and performing liquid fermentation at 25-35 ℃ for 3-5 days to obtain fermentation liquor;
4) preparing a fermentation preparation: adding an enzyme preparation into the fermentation liquor, and uniformly mixing to obtain the composite glycosyl biological feed leavening agent.
As a preferable embodiment of the above-mentioned means, in the step 2), the mass ratio of the sugar base to water is 1: 5.
The invention also claims the application of the composite glycosyl biological feed leaven prepared by the preparation method in preparing biological feed as the same inventive concept as the subject.
A method for preparing biological feed comprises the following steps:
and (3) crushing and softening silage raw materials, adding the compound glycosyl biological feed starter, and sealing and storing to obtain the biological feed.
As a preferable technical scheme of the technical scheme, the mass ratio of the silage to the compound glycosyl biological feed leavening agent is 100: 1; the time for sealed storage at 20-35 ℃ is 15 days.
As the preferable technical scheme of the technical scheme, the process of crushing and softening the silage raw materials comprises the following steps: cutting the raw materials into 2-3cm segments, bundling the segments, sealing with sealed bags, stacking, coating film, and fermenting at 20-35 deg.C for 7 days or longer.
According to the technical scheme, the composite glycosyl biological feed leaven provided by the invention can quickly reduce the pH value of biological feed and is convenient for quick fermentation and storage of the biological feed; and the raw materials are fermented by the leaven, so that the nutritive value of the biological feed can be obviously improved, the number of probiotics is increased, the content of tannin anti-nutritional factors in the biological feed is reduced, and the safety of the biological feed is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a diagram showing the change of pH during fermentation of broussonetia papyrifera leaves and straws;
FIG. 2 is a graph showing the change of tannin content in each group after fermentation of broussonetia papyrifera leaves;
FIG. 3 is a graph showing the content of lactic acid bacteria in each group after fermentation with broussonetia papyrifera leaves.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment of the invention discloses a compound glycosyl biological feed leaven and a preparation method and application thereof. The strains, sugars and enzyme components used in the examples are all commercially available.
Example 1
A compound glycosyl biological feed leaven comprises the following raw materials: 20 parts of sugar matrix, 2 parts of probiotics and 2 parts of enzyme preparation; wherein the compound sugar matrix comprises 9 parts of glucose, 3 parts of fructose, 2 parts of lactose, 3.8 parts of maltose and 2.2 parts of sucrose; the probiotics comprise 0.16 part of bacillus subtilis, 0.24 part of bacillus licheniformis, 0.46 part of lactobacillus plantarum, 0.34 part of lactobacillus fermentum, 0.1 part of streptococcus lactis, 0.56 part of saccharomyces cerevisiae and 0.14 part of bifidobacterium; the enzyme preparation comprises: 1.16 parts of cellulase and 0.84 part of tannase.
Example 2
A compound glycosyl biological feed leaven comprises the following raw materials: 20 parts of sugar matrix, 2 parts of probiotics and 2 parts of enzyme preparation; wherein the compound sugar matrix comprises 6 parts of glucose, 4 parts of fructose, 1 part of lactose, 5 parts of maltose and 4 parts of sucrose; the probiotics comprise 0.3 part of bacillus subtilis, 0.3 part of bacillus licheniformis, 0.4 part of lactobacillus plantarum, 0.3 part of lactobacillus fermentum, 0.1 part of streptococcus lactis, 0.4 part of saccharomyces cerevisiae and 0.2 part of bifidobacterium; the enzyme preparation comprises: 0.9 part of cellulase and 1.1 parts of tannase.
Example 3
A compound glycosyl biological feed leaven comprises the following raw materials: 20 parts of sugar matrix, 2 parts of probiotics and 2 parts of enzyme preparation; wherein the compound sugar matrix comprises 7.8 parts of glucose, 2.4 parts of fructose, 2.4 parts of lactose, 4.6 parts of maltose and 2.8 parts of sucrose; the probiotics comprise 0.2 part of bacillus subtilis, 0.14 part of bacillus licheniformis, 0.52 part of lactobacillus plantarum, 0.4 part of lactobacillus fermentum, 0.04 part of streptococcus lactis, 0.6 part of saccharomyces cerevisiae and 0.1 part of bifidobacterium; the enzyme preparation comprises: 0.98 parts of cellulase and 1.02 parts of tannase.
Example 4
A compound glycosyl biological feed leaven comprises the following raw materials: 20 parts of sugar matrix, 2 parts of probiotics and 2 parts of enzyme preparation; wherein the compound sugar matrix comprises 6.4 parts of glucose, 3.6 parts of fructose, 1.6 parts of lactose, 4.4 parts of maltose and 4 parts of sucrose; the probiotics comprise 0.1 part of bacillus subtilis, 0.2 part of bacillus licheniformis, 0.56 part of lactobacillus plantarum, 0.42 part of lactobacillus fermentum, 0.14 part of streptococcus lactis, 0.46 part of saccharomyces cerevisiae and 0.12 part of bifidobacterium; the enzyme preparation comprises: 1.2 parts of cellulase and 0.8 part of tannase.
Example 5
A compound glycosyl biological feed leaven comprises the following raw materials: 10 parts of sugar matrix, 4 parts of probiotics and 1.5 parts of enzyme preparation; wherein the compound sugar matrix comprises 4.5 parts of glucose, 1.2 parts of fructose, 1.4 parts of lactose, 1.5 parts of maltose and 1.2 parts of sucrose; the probiotics comprise 0.42 part of bacillus subtilis, 0.28 part of bacillus licheniformis, 0.89 part of lactobacillus plantarum, 0.78 part of lactobacillus fermentum, 0.32 part of streptococcus lactis, 0.97 part of saccharomyces cerevisiae and 0.34 part of bifidobacterium; the enzyme preparation comprises: 0.825 parts of cellulase and 0.675 parts of tannase.
Example 6
A compound glycosyl biological feed leaven comprises the following raw materials: 25 parts of sugar matrix, 0.5 part of probiotics and 0.2 part of enzyme preparation; wherein the compound sugar matrix comprises 10 parts of glucose, 3.5 parts of fructose, 2 parts of lactose, 5.5 parts of maltose and 4 parts of sucrose; the probiotics comprise 0.05 part of bacillus subtilis, 0.04 part of bacillus licheniformis, 0.15 part of lactobacillus plantarum, 0.075 part of lactobacillus fermentum, 0.05 part of streptococcus lactis, 0.1 part of saccharomyces cerevisiae and 0.035 part of bifidobacterium; the enzyme preparation comprises: 0.1 part of cellulase and 0.1 part of tannase.
Example 7
A compound glycosyl biological feed leaven comprises the following raw materials: 22 parts of sugar matrix, 1 part of probiotics and 0.6 part of enzyme preparation; wherein the compound sugar matrix comprises 8.9 parts of glucose, 2.8 parts of fructose, 2.5 parts of lactose, 4.7 parts of maltose and 3.1 parts of sucrose; the probiotics comprise 0.2 part of bacillus subtilis, 0.14 part of bacillus licheniformis, 0.52 part of lactobacillus plantarum, 0.4 part of lactobacillus fermentum, 0.04 part of streptococcus lactis, 0.6 part of saccharomyces cerevisiae and 0.1 part of bifidobacterium; the enzyme preparation comprises: 0.98 parts of cellulase and 1.02 parts of tannase.
Wherein, the preparation process of the sugar substrate in the examples 1 to 7 is as follows: mixing glucose, fructose, lactose, maltose and sucrose uniformly according to the weight part ratio, and packaging in an aseptic bag; the preparation process of the probiotics comprises the following steps: culturing each strain into bacterial liquid, mixing, centrifuging to obtain precipitate, adding normal saline, shaking, adding into sterile folium Broussonetiae powder, mixing, drying, and sealing; the enzyme preparation is prepared by mixing cellulase and tannase according to the mass ratio.
The preparation method of the composite glycosyl biological feed leaven comprises the following steps: firstly, adding sugar substrate and water into a liquid fermentation tank to prepare liquid sugar substrate with the ratio of 2: 1; and adding probiotics, performing liquid fermentation at 30 ℃ for 3 days, and adding an enzyme preparation before use to obtain the compound glycosyl biological feed leavening agent.
Example 5
After 100 parts of silage raw materials are crushed and softened, 1 part of compound glycosyl biological feed leaven is added, and the silage is sealed by a sealing bag and stored for 15 days to be used. The process of smashing and softening silage raw materials comprises the following steps: cutting folium Broussonetiae into 2-3cm segments, bundling, sealing with sealed bag, stacking, coating film, and fermenting at 20-35 deg.C for 7 days or longer.
Selecting folium Broussonetiae, and performing fermentation experiment according to the above method, wherein the blank group is only subjected to anaerobic fermentation at 20-35 deg.C for 15 days; no starter is added; pH of fermentation samples was measured using a solid pH meter as shown in FIG. 1, and after 15 days of fermentation, all groups had pH values between 4 and 5.
The tannin is measured according to the GB/T27985-2011 method, the tannin content in the paper mulberry leaves in the fermentation process is detected, as shown in figure 2, and as can be seen from figure 2, after 15 days of fermentation, the tannin content in all groups is reduced by about 40%.
The lactobacillus content detection method comprises the following steps:
according to the regulations in GB/T14699.1, when taking samples, the representativeness of the samples is taken, and the sampling process is carried out in an ultra-clean bench in a sterile mode.
Preparation of the immersion liquid: aseptically weighing 5g of fermented broussonetia papyrifera feed products in each group in an ultraclean workbench, placing the materials in a 50mL sterile centrifuge tube, adding 45mL of physiological saline, soaking for 4h at room temperature, and uniformly mixing by shaking in a vortex mixer to obtain the microorganism-containing immersion liquid.
Sucking 1mL of the mixed solution by a pipette, placing the solution in a 15mL sterile centrifuge tube, adding 9mL of normal saline, and mixing by vortex in a vortex mixer to obtain 10-1Concentration immersion; pipetting 10-1Placing 1mL of the concentration immersion liquid into a 15mL sterile centrifuge tube, adding 9mL of normal saline, and uniformly mixing by vortex in a vortex mixer to obtain a 10-2 concentration immersion liquid; thereby obtaining 10 respectively-3、10-4、10-5、10-6、10-7、10-8And (5) performing concentration immersion.
Sucking 10 μ L (or adding or subtracting according to actual requirement) of 10 μ L respectively with pipette-1、10-2、10-3、10-4、10-5、10-6、10-7、10-8Adding the concentration immersion liquid to MRS agar culture medium respectively, and coating with a coater uniformly; placing into a vacuum bag, sealing, removing air, making anaerobic environment, and culturing in a 30 deg.C constant temperature incubator; or alternatively adding CO at 30 deg.C2Anaerobic culture in an incubator; and culturing for 72 h.
Selecting a culture dish with the lactic acid bacteria colony number between 30 and 300CFU for counting, wherein the colony number of each concentration is determined by the average number of at least three plates; the results are shown in FIG. 3; as shown in FIG. 3, the number of lactic acid bacteria in all groups increased by about 120% after 15 days of fermentation.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.