CN105918618A - Solid microecological product rich in probiotics and preparation method and application thereof - Google Patents

Abstract

The invention provides a solid micro-ecological product rich in probiotics, the raw materials of which include Flammulina velutipes cultivation waste, bean cake powder, bran, corn flour, rice bran, magnesium sulfate, disodium hydrogen phosphate, ferric chloride, water, good Aerobic probiotics, facultative anaerobic and anaerobic probiotics. The invention also provides a preparation method of a solid-state micro-ecological product rich in probiotics, which includes the preparation of solid-state fermentation raw materials and the steps of fermenting the solid-state fermentation raw materials with probiotics. The solid micro-ecological product rich in probiotics has remarkable probiotic function, is easy to absorb, has rich nutrient content, can be used as feed, and the preparation method is simple and low in cost.

Description

Solid micro-ecological product rich in probiotics and its preparation method and application

technical field

The invention relates to the field of biotechnology, in particular to a solid micro-ecological product rich in probiotics and its preparation method and application.

Background technique

China is rich in edible fungi resources, and the output of edible fungi accounts for more than 70% of the world's total output. Among them, Flammulina velutipes is a very popular edible fungus in the market. Flammulina velutipes cultivation waste is the culture material of Flammulina velutipes cultivation. After the growth and metabolism of Flammulina velutipes, cellulose, hemicellulose and wood have been degraded to varying degrees. Compared with before fermentation, the protein and crude fat were significantly increased, and the crude cellulose was significantly reduced, and it was rich in amino acids, fungal polysaccharides, and trace elements such as Fe, Ca, Zn, and Mg. For example, in the cultivation waste of Flammulina velutipes, which is mainly made of cottonseed hulls, the content of crude protein is 10.03%, crude fiber is 18.74%, crude fat is 1.94%, and crude ash is 0.06%. However, many mushroom farmers or manufacturers use the waste after production as fuel or discard it anywhere, which is not only wasteful, but also seriously pollutes the environment.

Contents of the invention

The invention provides a kind of probiotic-rich solid micro-ecological product with significant probiotic function, easy absorption and rich nutritional content, prepared by using Flammulina velutipes cultivation waste as the main raw material and fermented by microorganisms, as well as its preparation method and application. above question.

The technical solution adopted in the present invention is: a solid micro-ecological product rich in probiotics, its raw materials include Flammulina velutipes cultivation waste, bean cake powder, bran, corn flour, rice bran, magnesium sulfate, disodium hydrogen phosphate, trichloride Iron, water and probiotics. Wherein, the cultivation waste of Flammulina velutipes is the cultivation material of Flammulina velutipes, through the growth and metabolism process of Flammulina velutipes, the cellulose, hemicellulose and wood in the medium have been degraded to varying degrees, and the crude protein and crude fat are more Significantly increased before fermentation, crude cellulose was significantly reduced, and rich in amino acids, fungal polysaccharides and trace elements such as Fe, Ca, Zn, Mg.

Based on the above, the solid micro-ecological product rich in probiotics includes the following mass ratio and inoculation amount of raw materials: Flammulina velutipes cultivation waste and bean cake powder, 1% ~5% bran, 1%~3% corn flour, 1%~3% rice bran, 0.1%~1% magnesium sulfate, 0.1%~1% disodium hydrogen phosphate, 0.1%~1% Ferric chloride and 30% to 40% water; the inoculation amount is 2% to 5% of probiotics.

Based on the above, the probiotics are one or a combination of aerobic probiotics, anaerobic probiotics and facultative anaerobic probiotics.

Based on the above, the aerobic probiotics are one or a combination of Bacillus colloidus, Bacillus subtilis and Bacillus natto.

Based on the above, the facultative anaerobic or anaerobic probiotics are one or a combination of Enterococcus faecalis, Pediococcus pentosaceae, Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium, Clostridium butyricum.

The present invention also provides a method for preparing the above-mentioned solid micro-ecological product rich in probiotics, comprising the following steps:

Crush the cultivation waste of Flammulina velutipes into particles with a particle size of 20 to 100 meshes, and then add bean cake powder, 1% to 5% of bran, 1% to 3% of the weight of the waste of Flammulina velutipes into it corn flour, 1%~3% rice bran, 0.1%~1% magnesium sulfate, 0.1%~1% disodium hydrogen phosphate, 0.1%~1% ferric chloride and 30%~40% water and mixed to prepare solid-state fermentation raw materials;

Probiotics are added to the solid-state fermentation raw material according to the inoculum amount of 2% to 5%, and fermentation treatment is carried out, so as to obtain the solid micro-ecological product rich in probiotics.

Based on the above, the fermentation treatment step includes using the aerobic probiotics to perform aerobic fermentation for 2-7 days and using the anaerobic probiotics or the facultative anaerobic probiotics to perform anaerobic fermentation treatment.

The present invention also provides an application of the above-mentioned solid micro-ecological product rich in probiotics as animal feed.

The present invention provides the probiotic-rich solid micro-ecological product with rich nutritional components and reasonable proportions, and the division of work among the probiotics is clear, and the cellulose, protein, antigen and anti-nutritional components in the solid-state fermentation raw materials can be Factors are efficiently degraded into small molecular proteins and small peptides, which improves the utilization rate of proteins. At the same time, probiotics will produce a large number of oligopeptides, glutamic acid, lactic acid, vitamins and other substances, so as to obtain significant probiotic functions, easy to absorb, and active small peptides Solid micro-ecological products rich in protein and nutrients rich in probiotics.

Specifically, the solid-state fermentation raw material prepared from Flammulina velutipes cultivation waste as the main raw material is rich in crude protein, crude fiber, crude fat, crude ash, amino acids, fungal polysaccharides and trace elements such as Fe, Ca, Zn, Mg, etc., providing Rich source of nutrition.

Bacillus colloidus is a nitrogen-fixing bacterium that can fix nitrogen in the atmosphere into the solid-state fermentation raw material, and then convert nitrogen into protein through other probiotics, thereby increasing the protein content.

Subtilisin, polymyxin, nystatin, gramicidin and other active substances produced during the growth of Bacillus subtilis, these active substances have obvious effects on pathogenic bacteria or opportunistic pathogenic bacteria of endogenous infection Inhibitory effect; at the same time, Bacillus subtilis can quickly consume free oxygen in the intestinal tract, causing intestinal hypoxia, promoting the growth of beneficial anaerobic bacteria, and producing organic acids such as lactic acid, reducing the pH value of the intestinal tract, and indirectly inhibiting the growth of other pathogenic bacteria It can also stimulate the growth and development of animal immune organs, activate T and B lymphocytes, increase the level of immunoglobulin and antibodies, enhance cellular immunity and humoral immunity, and improve group immunity. It can synthesize α-amylase, protease, Enzymes such as lipase and cellulase play a role together with digestive enzymes in animals in the digestive tract; they can also synthesize vitamin B1, B2, B6, niacin and other B vitamins to increase interferon and macrobiotics in animals. Phage activity.

Bacillus natto is a kind of beneficial bacteria with acid-resistant and heat-resistant characteristics. It has a 100% survival rate in four hours under gastric acid. It also has a strong ability to inhibit pathogenic bacteria. It is the best environmental tolerance among various beneficial bacteria. One of the strains that reach the small intestine, after oral administration, it can change the ecology of the intestinal flora, help normalize the function of the digestive tract, make defecation smooth, and maintain the physiological environment in the body. At the same time, it can also produce acid, regulate the intestinal flora, enhance the immune response of animal cells, and can generate a variety of proteases (especially alkaline protease), glucoamylase, lipase, amylase, which can degrade some complex substances in plants. of carbohydrates.

Enterococcus faecalis is one of the main flora in the intestinal tract of humans and animals. It can produce natural antibiotics, which are beneficial to the health of the body. At the same time, it can also produce antibacterial substances such as bacteriocins, inhibit the growth of pathogenic bacteria such as E. environment; it can also inhibit the reproduction of urease-producing bacteria and spoilage bacteria in the intestine, reduce the content of urease and endotoxin in the intestine, and reduce the content of ammonia and endotoxin in the blood; at the same time, Enterococcus faecalis can decompose protein into small peptides, Synthesize B vitamins, enhance the activity of macrophages, promote the immune response of animals, and increase the level of antibodies. In addition, Enterococcus faecalis can also decompose some proteins into amides and amino acids, and convert most of the nitrogen-free extracts of carbohydrates into L-type lactic acid, which can synthesize L-calcium lactate for calcium and promote the absorption of calcium; Soften the fiber and increase the conversion rate of the fiber.

Lactobacillus plantarum has many health effects, such as certain immune regulation, inhibitory effect on pathogenic bacteria, lowering serum cholesterol levels and preventing cardiovascular diseases, maintaining intestinal flora balance, promoting nutrient absorption, alleviating lactose Intolerance, inhibition of tumor cell formation, etc.

Lactobacillus acidophilus can adjust the balance of intestinal flora, inhibit the proliferation of harmful intestinal microorganisms, and can also secrete antibiotics, such as acidophilus, acidophilus and lactobacillus, which are effective against intestinal pathogenic bacteria. produced antagonism.

Bifidobacteria can produce lactic acid and acetic acid after fermentation, which can improve the utilization rate of calcium, phosphorus and iron, and promote the absorption of iron and vitamin D. At the same time, bifidobacteria can produce essential nutrients such as vitamins B1, B2, B6, B12, alanine, valine, aspartic acid and threonine.

Clostridium butyricum can tolerate gastric acid entering the intestinal tract, secrete butyric acid, an important nutrient substance for intestinal mucosal regeneration and repair, and can promote the growth of intestinal beneficial bacteria such as Bifidobacterium, inhibit the growth of intestinal harmful bacteria such as Shigella dysenteriae, and restore Balance the intestinal flora, reduce the production of amine, ammonia, indole and other intestinal toxins and poison the intestinal mucosa, restore intestinal immune function and normal physiological functions, and inhibit the production of inflammatory factors such as IL-8 and TNF-α Excessive abnormal expression, inhibiting the overexpression of anti-colon antibody IgG, reducing the conversion rate of B lymphocytes, increasing the conversion rate of T lymphocytes, correcting intestinal immune disorders, restoring intestinal immune tolerance, and eliminating inflammation and ulcers. At the same time, Clostridium butyricum produces enzymes and vitamin-like beneficial substances in the intestinal tract to promote the digestion and absorption of nutrients.

The present invention fixes nitrogen into the solid-state fermentation raw material with Flammulina velutipes cultivation waste as raw material through microbial fermentation and converts it into protein, and utilizes the enzymolysis of various proteases secreted by probiotics during the growth and development process to decompose the macromolecular protein into small molecular protein and small peptide, and convert the solid-state fermentation raw material into a solid micro-ecological product rich in probiotics with high value; The type and quantity of bacteria are significantly increased, thereby better inhibiting the growth and reproduction of harmful microorganisms such as E. Disease prevention and disease resistance. Because the solid micro-ecological products rich in probiotics and their preparation methods and applications have a unique aroma, they can greatly improve the flavor and quality of the feed, increase the feed intake and appetite of the raised animals, and thus promote better growth of the animals Development, significantly shorten the feeding cycle. The method has simple preparation method and low cost.

Description of drawings

Fig. 1 is the gel diagram of antigen protein decomposition under different fermentation time.

detailed description

The technical solutions of the present invention will be described in further detail below through specific implementation methods.

Example 1

The present embodiment provides a kind of solid micro-ecological product rich in probiotics, specifically comprising raw materials of the following quality: 100 Kg of Flammulina velutipes cultivation waste, 1 Kg of bean cake powder, 1 Kg of bran, 1 Kg of corn flour, 1 Kg of rice bran, sulfuric acid Magnesium 0.1 Kg, disodium hydrogen phosphate 0.1 Kg, ferric chloride 0.1 Kg, 30 Kg of water, 2.74Kg of aerobic probiotics composed of Bacillus colloidus, Bacillus subtilis and Bacillus natto; 2.74Kg of Facultative anaerobic and anaerobic probiotics consisting of Enterococcus faecalis, Pediococcus pentosaceae, Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium, Clostridium butyricum.

This embodiment also provides a method for preparing the above-mentioned solid micro-ecological product rich in probiotics, comprising the following steps:

100 Kg of Flammulina velutipes cultivation waste is crushed into particles with a particle size of 100 mesh, and then it is mixed with 1 Kg of bean cake powder, 1 Kg of bran, 1 Kg of corn flour, 1 Kg of rice bran, 0.1 Kg of magnesium sulfate, 0.1 Kg of disodium hydrogen phosphate, three 0.1 Kg of ferric chloride and 30 Kg of water are mixed to prepare solid-state fermentation raw materials;

Add 2.74Kg of aerobic probiotics composed of Bacillus colloidus, Bacillus subtilis and Bacillus natto to the solid-state fermentation raw material; Facultative anaerobic and anaerobic probiotics composed of Lactobacillus acidum, Bifidobacterium, and Clostridium butyricum; aerobic solid-state fermentation was carried out in an aerobic fermentation tank for 3 days; Anaerobic fermentation is carried out in a one-way membrane anaerobic bag, thereby preparing the solid micro-ecological product rich in probiotics.

The probiotic-rich solid microecological product prepared in this example can be used as pig feed.

Example 2

The embodiment of the present invention provides a solid micro-ecological product rich in probiotics. The specific raw material composition is roughly the same as that in Example 1. The difference is that the quality of each raw material is different. The quality of each raw material in this embodiment is:

Flammulina velutipes cultivation waste 100 Kg, bean cake powder 10 Kg, bran 5 Kg, corn flour 3 Kg, rice bran 3 Kg, magnesium sulfate 1 Kg, disodium hydrogen phosphate 1 Kg, ferric chloride 1 Kg, 40 Kg of water, 3.35 Kg of aerobic probiotics composed of Bacillus colloidus, Bacillus subtilis and Bacillus natto and 3.35Kg of anaerobic probiotics composed of Enterococcus faecalis, Pediococcus pentosaceae and Lactobacillus plantarum.

The preparation method of the solid micro-ecological product rich in probiotics provided in this example is the same as the preparation method in Example 1.

The probiotic-rich solid microecological product prepared in this example can be used as pig feed.

Example 3

The embodiment of the present invention provides a solid micro-ecological product rich in probiotics. The specific raw material composition is roughly the same as that in Example 1. The difference is that the quality of each raw material is different. The quality of each raw material in this embodiment is:

Flammulina velutipes cultivation waste 100 Kg, bean cake powder 5 Kg, bran 3 Kg, corn flour 2 Kg, rice bran 2 Kg, magnesium sulfate 0.5 Kg, disodium hydrogen phosphate 0.5 Kg, ferric chloride 0.5 Kg, 35 Kg of water, 7.82 Kg of aerobic probiotics composed of Bacillus coliformis and Bacillus subtilis.

The preparation method for preparing the probiotic-rich solid micro-ecological product provided in this example is the same as the preparation method in Example 1.

The probiotic-rich solid microecological product prepared in this example can be used as pig feed.

Test Testing and Verification

(1) Antigen detection

Take 1.0 g of the solid-state probiotic-rich probiotic product prepared through different fermentation times in Example 1, add 20.00 ml pH=8.0 detection reagent, soak at room temperature for 1 hour, and stir once every 10 minutes. Wherein, the detection reagent is Tris-HCl with a concentration of 0.03 mol/L and a concentration of 0.01 mol/L β-mercaptoethanol mixed composition. After soaking, centrifuge at a centrifugal speed of 10000rpm for 20 minutes, take 80 μL of the supernatant, add 20 μL of blue sample buffer (5×) to the boiling water bath for 3 minutes, and then take 10 μL of each sample solution and add them to the gel tank In the process, turn on the voltage regulator power switch, set the voltage to 180v for electrophoresis, and end the electrophoresis when the blue band of the sample buffer migrates to the bottom of the gel, carefully peel off the gel with an offset plate, and put it into a container with staining solution , after staining for 10 min, pour off the staining solution, add decolorization solution for decolorization, until the back color of the gel is the initial white and the bands are clearly visible.

The decomposition of the antigenic protein is shown in Figure 1. It can be seen from the gel pattern that the antigenic protein was decomposed to a large extent when the fermentation time was 3 days compared with the unfermented raw material, and when the fermentation time was 7 days, a larger amount was obtained. The degree of decomposition, compared with the unfermented material, the band of the antigen is very weak. The results show that the fermentation of probiotics can effectively decompose protein antigens and anti-nutritional factors in raw materials, and at the same time convert a large amount of protein into active small peptide proteins, thereby effectively improving protein utilization.

(2) Detection of crude protein in medium

Take 1.0 g of the probiotic-rich solid-state micro-ecological product prepared through different fermentation times in Example 1, and use the Kjeldahl method to detect the crude protein content in different samples with reference to the provisions of GB/T6432-94.

The test results show that the crude protein content of the dried and prepared unfermented raw materials is 10.02%, and the moisture content is 11.67%; the crude protein content of the solid probiotic-rich micro-ecological products dried after 3 days of fermentation is 13.26%, The moisture content is 11.64%, and the crude protein content is 3.24% higher than that of unfermented raw materials; the crude protein content in the probiotic-rich solid micro-ecological products dried after 7 days of fermentation is 15.13%, and the moisture content is 11.69%, The crude protein content is 5.11% higher than that of unfermented raw materials.

It can be seen that, through the fermentation of probiotics, the protein content in the cultivation waste of Flammulina velutipes can be significantly increased.

(3) Animal feeding test

For the test, 40 young piglets were randomly divided into 4 groups, including a blank control group and 3 test groups, with 10 male and half female piglets in each group. The pigs in the blank control group were fed with common grain and common feed, and the pigs in the test group were fed with the probiotic-rich solid-state microecological products prepared in Examples 1 to 3. The pigs had free access to food and water, and were immunized and dewormed as usual. During the test period, observe and record the swine flu situation and the mental state of the pigs every day, and the test period is 90 days. During the feeding process, statistics were made on the development and incidence of pigs in each group. At the end of the feeding experiment, SPSS19.0 was used for the experimental data of the growth performance and immune indexes of pigs in each group. Statistical software for statistical analysis.

The growth performance indicators of 40 young piglets were measured, and the results are shown in Table 1. It can be seen that the daily feed intake, daily weight gain and feed-to-meat ratio of the three test groups were better than those of the control group.

Table 1 Growth performance indicators

project control group Test 1 group Test 2 groups Test 3 groups Daily feed intake /kg 1.05±0.06 1.20±0.05 1.65±0.10 1.68±0.07 Daily gain/kg 0.30±0.01 0.60±0.05 0.63±0.03 0.65±0.02 Feed to meat ratio 3.50±0.02 2.00±0.01 2.62±0.02 2.58±0.03

At 8:00 in the morning on the last day of the test period, each group selected two ends to measure its IgA, IgG and IgM levels, and the immune indexes were measured. The results are shown in Table 2. It can be seen that in the serum of the 33 test groups The levels and incidence of IgA, IgG and IgM in the patients were lower than those in the control group.

Table 2 Immune indicators and incidence

project control group Test 1 group Test 2 groups Test 3 groups IgA/(g/L) 0.12±0.03 0.21±0.03 0.19±0.04 0.22±0.01 IgG/(g/L) 6.00±0.67 8.55±1.21 8.12±0.87 8.87±1.13 IgM (g/L) 1.04±0.15 1.35±0.20 1.32±0.22 1.38±0.10 Morbidity 12.2% 2.5% 3.2% 2.0%

It can be seen that the probiotic-rich solid-state micro-ecological products produced by the present invention with Flammulina velutipes cultivation waste as the main raw material are used as feed, which can promote the increase of the feed intake of the raised animals and significantly enhance their appetite, thereby promoting better health of the animals. Growth and development, significantly shorten the feeding cycle.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them; although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that: the present invention can still be The specific implementation of the invention is modified or some technical features are equivalently replaced; without departing from the spirit of the technical solution of the present invention, they should be included in the scope of the technical solution claimed in the present invention.

Claims (8)

1. the solid-state Tiny ecosystem goods rich in probiotic bacteria, it is characterised in that its raw material includes golden mushroom plantation waste material, soybean cake powder, wheat bran, Semen Maydis powder, Testa oryzae, magnesium sulfate, disodium hydrogen phosphate, ferric chloride, probiotic bacteria and water.

Solid-state Tiny ecosystem goods rich in probiotic bacteria the most according to claim 1, it is characterized in that, it includes following quality proportioning and the raw material of inoculum concentration: golden mushroom plantation waste material and account for respectively described golden mushroom plantation waste material weight 1%～10% the ferric chloride of the disodium hydrogen phosphate of the magnesium sulfate of the Testa oryzae of the Semen Maydis powder of the wheat bran of soybean cake powder, 1%～5%, 1%～3%, 1%～3%, 0.1%～1%, 0.1%～1%, 0.1%～1% and the water of 30%～40%；Inoculum concentration is the probiotic bacteria of 2%～5%.

Solid-state Tiny ecosystem goods rich in probiotic bacteria the most according to claim 1 and 2, it is characterised in that described probiotic bacteria is the combination of one or more in aerobic probiotic bacteria, anaerobism probiotic bacteria and amphimicrobian probiotic bacteria.

Solid-state Tiny ecosystem goods rich in probiotic bacteria the most according to claim 3, it is characterised in that described aerobic probiotic bacteria is the combination of one or more in colloid bacillus cereus, bacillus subtilis and Bafillus natt.

Solid-state Tiny ecosystem goods rich in probiotic bacteria the most according to claim 4, it is characterized in that, described amphimicrobian or anaerobism probiotic bacteria are the combination of one or more in enterococcus faecalis, Pediococcus pentosaceus, Lactobacillus plantarum, bacillus acidophilus, bacillus bifidus, clostridium butyricum.

6. a preparation method for the solid-state Tiny ecosystem goods rich in probiotic bacteria described in any one of Claims 1 to 5, comprises the following steps:

Golden mushroom plantation waste powder is broken into the granule that granularity is 20～100 mesh, be added thereto to the most respectively to account for described golden mushroom plantation waste material weight 1%～10% the ferric chloride of the disodium hydrogen phosphate of the magnesium sulfate of the Testa oryzae of the Semen Maydis powder of the wheat bran of soybean cake powder, 1%～5%, 1%～3%, 1%～3%, 0.1%～1%, 0.1%～1%, 0.1%～1% and the water of 30%～40% mixing, be configured to solid fermentation raw material；

It is 2%～5% to add probiotic bacteria carry out fermentation process according to inoculum concentration in described solid fermentation raw material, thus prepares the described solid-state Tiny ecosystem goods rich in probiotic bacteria.

The preparation method of the solid-state Tiny ecosystem goods rich in probiotic bacteria the most according to claim 6, it is characterized in that, described fermentation process step includes using described aerobic probiotic bacteria to carry out aerobic fermentation 2～carrying out anaerobic fermentation process with the described anaerobism probiotic bacteria of employing or described amphimicrobian probiotic bacteria in 7 days.

8. the application of the solid-state Tiny ecosystem goods rich in probiotic bacteria described in an any one of Claims 1 to 5, it is characterised in that it is applied as animal feed.