CN113367230A - High-digestibility pediococcus acidilactici fermented wet soybean meal and preparation method thereof - Google Patents

High-digestibility pediococcus acidilactici fermented wet soybean meal and preparation method thereof Download PDF

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CN113367230A
CN113367230A CN202110416893.8A CN202110416893A CN113367230A CN 113367230 A CN113367230 A CN 113367230A CN 202110416893 A CN202110416893 A CN 202110416893A CN 113367230 A CN113367230 A CN 113367230A
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黄竹
丁毅
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Fengmei Shenyang Biological High Tech Industry Research Institute Co ltd
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Abstract

The invention relates to the field of animal feed raw materials, in particular to wet soybean meal fermented by pediococcus acidilactici with high digestibility and a preparation method thereof. Crushing the soybean meal raw material, adding water, uniformly mixing, sterilizing, simultaneously adding the liquid proliferated pediococcus acidilactici and the complex enzyme preparation, carrying out constant-temperature slow enzymolysis fermentation, carrying out solid-liquid separation after fermentation, drying and crushing to obtain the pediococcus acidilactici high-digestibility fermented wet soybean meal. By adding the method of combining the complex enzyme preparation and the pediococcus acidilactici mixed fermentation, the content of the acid soluble protein is up to 10.25 percent, the essential amino acid is integrally improved, and the digestibility is up to 79.68 percent. The method has the advantages that the nutritional contents of other various ingredients are increased, and the method plays an important role in improving the nutritional value of the soybean meal. Through the composition and the addition of the specific complex enzyme preparation and the synergistic effect among various enzymes, the content of non-starch polysaccharide is effectively reduced, and then the non-starch polysaccharide is fermented and utilized by microorganisms to form a series of metabolites which are beneficial to digestion and absorption, so that the digestion and utilization rate of the feed is improved.

Description

High-digestibility pediococcus acidilactici fermented wet soybean meal and preparation method thereof
Technical Field
The invention relates to the field of animal feed raw materials, in particular to wet soybean meal fermented by pediococcus acidilactici with high digestibility and a preparation method thereof.
Background
The soybean meal is used as a vegetable feed protein raw material and has received wide application and attention. However, the bean pulp contains more anti-nutritional factors, which seriously influences the digestion, absorption and utilization rate of the bean pulp in animal feed. The anti-nutritional factors in the soybean meal are divided into two types, namely heat-instability anti-nutritional factors and heat-stability anti-nutritional factors, wherein the heat-stability anti-nutritional factors mainly comprise soybean antigen protein globulin and beta-conglycinin, soybean oligosaccharides mainly comprise raffinose and stachyose, and the anti-nutritional factors also comprise trypsin inhibitor, phytic acid, urease activity and the like. The soybean antigenic proteins are mainly glycinin and beta-conglycinin, and have antigenicity and sensitization. After the antigen protein enters the animal body. The immune system is particularly sensitive to antigen proteins, causes allergic reactions, and is easy to cause diarrhea, reduce the production performance and even die in lactating animals, and simultaneously, the damage of intestinal mucosa and the reduction of the digestive absorption capacity of the intestinal tract are accompanied. Oligosaccharide substances in the soybean meal are another anti-nutritional factor, and the sugar is fermented and utilized by beneficial bacteria in intestinal tracts, but the content is too high, the flatulence can be caused by excessive fermentation gas production, and simultaneously, the diarrhea can be caused by fermentation products. In addition, the soybean meal also contains a certain amount of non-starch polysaccharide. Non-starch polysaccharides (NSP) are the main components of feed fibers, which enclose the feed nutrients inside the cell walls, part of the fibers being soluble in water and producing a sticky mass. These viscous materials inhibit the normal digestive function of the animal and prevent the animal from absorbing nutrients. If these NSPs are removed, nutrients can be released from the cell wall, thereby improving the metabolic energy and protein utilization. The non-starch polysaccharide in the soybean meal can be released by removing starch and protein surrounded by cell structures in the soybean meal by using an enzyme preparation and a fermentation mode, so that the metabolic energy of the soybean meal and the utilization rate of the protein are improved.
At present, a great number of patents about fermented soybean meal exist, and the reduction of the content of antigen protein and the content of oligosaccharide is mainly performed. However, there are few reports on the removal of non-starch polysaccharides from soybean meal. The digestion, absorption and utilization rate of the soybean meal in animal feed is seriously influenced.
The related patent, a high protein fermented soybean meal and application thereof show that although fermented soybean meal with high crude protein content can be obtained, the digestion and utilization rate of protein can be reduced due to limited protein absorption capacity of a body and the existence of other anti-nutritional factors. In summary, the content of non-starch polysaccharide and oligosaccharide can be reduced by microbial fermentation technology and enzyme engineering technology, the utilization rate of the soybean meal by animals can be improved by the accumulation of beneficial metabolites, and the position of the soybean meal in an animal protein feed source can be further improved.
Disclosure of Invention
The invention aims to solve the defects in the prior art, and the invention aims to provide wet soybean meal fermented by pediococcus acidilactici with high digestibility and a preparation method thereof.
In order to achieve the purpose, the invention mainly provides the following technical scheme:
a wet soybean meal fermented by Pediococcus acidilactici with high digestibility is prepared by adding Pediococcus acidilactici proliferated in liquid state and complex enzyme preparation into pulverized soybean meal raw material, and fermenting.
The method specifically comprises the following steps: crushing the soybean meal raw material, adding water, uniformly mixing, sterilizing, simultaneously adding the liquid proliferated pediococcus acidilactici and the complex enzyme preparation, carrying out constant-temperature slow enzymolysis fermentation, carrying out solid-liquid separation after fermentation, drying and crushing to obtain the pediococcus acidilactici high-digestibility fermented wet soybean meal. The mass ratio of the soybean meal raw material to water is 5-7: 3-5, wherein the conditions of constant-temperature slow enzymolysis and fermentation are 37 ℃ and 72 hours.
The complex enzyme preparation, namely the acidic protease, the neutral protease and the alkaline protease, comprises the following components in parts by mass: 2: 4, mixing.
The inoculation amount of the pediococcus acidilactici is 5-15%.
The addition amount of the acidic protease, the neutral protease and the alkaline protease is 2500U.
The pediococcus acidilactici (pediococcus acidilactici) is derived from a sample collected in a manual pickled Chinese cabbage workshop, is preserved in China general microbiological culture Collection center (CGMCC) at 9 and 16 days 2020, and has a preservation unit address as follows: no. 3 Xilu No. 1 Beijing, Chaoyang, and the preservation number is CGMCC No. 20658.
5. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 7, wherein: the pediococcus acidilactici proliferated in liquid state is prepared by inoculating pediococcus acidilactici preserved in a microorganism strain preservation center into an MRS liquid culture medium according to the inoculation amount of 1 ‰, culturing at 37 deg.C for 24 hr, and activating for 2-3 generations.
Crushing the soybean meal raw material, adding water, uniformly mixing, sterilizing, simultaneously adding the pediococcus acidilactici and the compound protease which are proliferated in a liquid state, fermenting at constant temperature at a low speed, performing solid-liquid separation after fermentation, drying and crushing to obtain the pediococcus acidilactici high-digestibility fermented wet soybean meal.
Due to the adoption of the technical scheme, the invention has the following advantages:
the content of acid soluble protein of the product is up to 10.25 percent by a method of adding a complex enzyme preparation and mixing and fermenting pediococcus acidilactici, the essential amino acid is integrally improved, and the digestibility is up to 79.68 percent. The method has the advantages that the nutritional contents of other various ingredients are increased, and the method plays an important role in improving the nutritional value of the soybean meal.
The non-starch polysaccharide content is effectively reduced by the composition and the adding amount of the specific complex enzyme preparation and the synergistic effect among various enzymes, and then the non-starch polysaccharide is fermented and utilized by microorganisms to form a series of metabolites which are beneficial to digestion and absorption, so that the digestion and utilization rate of the feed is improved.
And by adding a complex enzyme preparation and pediococcus acidilactici, the degradation products of the non-starch polysaccharides are further subjected to enzymolysis for biotransformation, beneficial metabolites are accumulated, the flavor of the fermented soybean meal is improved, and the obtained high-digestibility fermented soybean meal has better palatability.
The content of crude protein in the high-digestibility fermented soybean meal of the product is improved to 26.4 percent, the content of acid soluble protein is up to 10.25 percent, the content of essential amino acid is integrally improved, and the digestibility is up to 79.68 percent.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all 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.
Example one
Pulverizing two parts of 1kg soybean meal into 40 mesh, mixing with water at a ratio of 50%, sterilizing at 121 deg.C, using one part as test group and one part as control group, adding complex enzyme preparation (acid protease: neutral protease: alkaline protease: 4: 2: 4), and adding 2500U. The control group is not added with strains, the test group is added with 10% activated pediococcus acidilactici to carry out fermentation treatment (the pediococcus acidilactici is inoculated into MRS liquid culture medium according to the inoculation amount of 1 per mill, cultured for 24h at 37 ℃, activated for 3 generations), solid-liquid separation is carried out after 72 hours of fermentation, and drying and crushing are carried out.
Pediococcus acidilacticiii (Pediococcus acidilacticiii) is obtained from manual pickled Chinese cabbage workshop collected samples, is preserved in China general microbiological culture Collection center (CGMCC) at 9 and 16 days 2020, and the preservation unit address is as follows: no. 3 Xilu No. 1 Beijing, Chaoyang, and the preservation number is CGMCC No. 20658.
Table one comparison of the soybean meal raw material with the high digestibility pediococcus acidilactici fermented wet soybean meal of this example before and after treatment is as follows:
nutritional index Control group Test group
Crude protein 22.4％ 26.34％
Acid soluble protein 1.25％ 10.25％
Rate of digestion 43.52％ 79.72％
Total acid 0.6％ 4.8％
Coarse fiber 6.3％ 5.2％
Acid soluble protein/crude protein 5.58 38.9
Non-starch polysaccharides 13.99％ 9.99％
Content of beneficial viable bacteria — 6.8×108cfu/g
And (3) analysis: the table shows the analysis of the nutritional components of the soybean meal raw material control group of this example and the test group products obtained by the method of this example. The results show that the nutritional ingredients in the soybean meal are improved, wherein the protein content is increased from about 22% to about 26%, in addition, the macromolecular protein in the fermented soybean meal obtained by the implementation is obviously degraded into small molecular protein, namely acid-soluble protein, the acid-soluble protein content is up to 10.25%, and compared with the macromolecular protein, the molecular protein is more beneficial to the absorption of animals and promotes the development of intestinal tracts. The content of viable beneficial bacteria reaches 108The feed quality is improved by more than cfu/g and the digestibility is as high as 79.68 percent.
Change in essential amino acid composition in soybean meal before and after the epi-II treatment (%)
Figure RE-GDA0003175187300000051
Figure RE-GDA0003175187300000061
And (3) analysis: from the second table, the essential amino acid content is improved, and the amino acid balance of the soybean meal is further improved.
Variation of the content of epi-III antinutritional factors
Figure RE-GDA0003175187300000062
Figure RE-GDA0003175187300000071
From the third table, the anti-nutritional factor content is effectively reduced, and the nutritional value of the soybean meal is further improved.
Example two
Three parts of 1kg of soybean meal are crushed into 40 meshes as raw materials and are mixed with water according to the proportion of 50%, the mixture is sterilized at 121 ℃, one part is used as a test I group, the other part is used as a test II group, the other part is used as a control group, and three groups are added with a compound enzyme preparation, wherein the adding amount of the compound enzyme preparation (acidic protease, neutral protease and alkaline protease are 4: 2: 4) is 2500U. The control group is not added with strains, the test group I is added with 10% of activated pediococcus acidilactici for fermentation treatment, and the test group II is added with 10% of activated saccharomyces cerevisiae for fermentation treatment (the activation step of the saccharomyces cerevisiae is the same as that of the pediococcus acidilactici, and the strains are laboratory separation strains), and solid-liquid separation, drying and crushing are carried out after fermentation for 72 hours.
The following comparisons of conventional nutritional indicators for soybean meal feedstock and test group I and test group II are shown below:
Figure RE-GDA0003175187300000072
Figure RE-GDA0003175187300000081
and (3) analysis: as can be seen from the table four, the nutritional indexes in the soybean meal of the test group I and the test group II are improved, but the improvement effect of the test group I is better than that of the test group II,
table five changes in essential amino acid composition in soybean meal (%)
Figure RE-GDA0003175187300000082
Figure RE-GDA0003175187300000091
And (3) analysis: as can be seen from the table five, the content of the essential amino acid in the test group is improved compared with that in the control group, and the increase of the essential amino acid in the test group I is higher than that in the test group II, so that the soybean meal amino acid balance improvement effect after the fermentation treatment of the pediococcus acidilactici is better.
TABLE VI Change in antinutritional factor content
Anti-nutritional factor Control group Test group I Reduction amount Test group II Reduction amount
Trypsin inhibitor Activity (IU/g) 3.2 1.7 46.8％ 2.6 18.7％
Glycine max semispherin 7s (ng/g) 1.04 0.35 47.5％ 0.85 18.3％
Glycinin 11s (ng/g) 1.47 0.34 76.8％ 0.96 34.7％
Phytic acid content (mg) 5.46 1.10 79.8％ 3.98 27.1％
And (3) analysis: in table six, compared with the control group, the content of the anti-nutritional factors in the test group I and the test group II is reduced, but the content of the anti-nutritional factors in the test group I is reduced more, and the nutritional value of the soybean meal is improved more.
Implementation III
Pulverizing four parts of 1kg soybean meal into 40 mesh as raw material, mixing with water at a ratio of 50%, sterilizing at 121 deg.C, one part as test I group, one part as test II group, one part as test III group, and one part as control group,
the four groups are added with complex enzyme preparation, wherein the addition amount of the complex enzyme preparation (acid protease: neutral protease: alkaline protease is 4: 2: 4) is 2500U. The control group was not treated with added strains, and then test group I was fermented by adding 5% of activated pediococcus acidilactici, test group II was fermented by adding 10% of activated pediococcus acidilactici, and test group III was fermented by adding 15% of activated pediococcus acidilactici. Fermenting for 72 hours, separating solid and liquid, drying and crushing.
The comparison of the conventional nutritional indexes of the raw material of the soybean meal and the test treatment groups is as follows:
nutritional index Control group Test group I Test group II Test group III
Crude protein 22.4％ 24.32％ 26.34％ 25.46％
Acid soluble protein 1.25％ 5.26％ 10.25％ 6.47％
Rate of digestion 43.52％ 52.34％ 79.72％ 61.25％
Total acid 0.6％ 1.4％ 4.8％ 2.7％
Coarse fiber 6.3％ 5.8％ 5.2％ 6.0％
Acid soluble protein/crude protein 5.58 21.6 38.9 25.4
Non-starch polysaccharides 13.99％ 12.78％ 9.99％ 12.12％
Content of beneficial viable bacteria — 7.6×105cfu/g 6.8×108cfu/g 7.4×107cfu/g
And (3) analysis: according to the seventh table, compared with the control group, the soybean meal related nutritional index of the test group II is improved to a better extent, so that the nutritional index of the fermented soybean meal is improved most obviously by the 10% pediococcus acidilactici treatment group.
TABLE eight changes in essential amino acid composition in the soybean meal (%)
Figure RE-GDA0003175187300000101
Figure RE-GDA0003175187300000111
And (3) analysis: as can be seen from the table five, the content of the essential amino acid in the test group is improved compared with that in the control group, and the increase of the essential amino acid in the test group II is higher than that in the test group I and the test group III, so that the soybean meal amino acid balance improvement effect after the fermentation treatment of the pediococcus acidilactici with the addition amount of 10% is better.
(TABLE ninth) variation in anti-nutritional factor content
Figure RE-GDA0003175187300000121
And (3) analysis: the ninth table shows that, compared with the control group, the anti-nutritional factor content of the test group I, the test group II and the test group III all decrease, but the anti-nutritional factor content of the test group II decreases more than that of the control group, the test group I and the test group III, so that the nutritional value of the soybean meal of the test group II is improved more.
The above inspection method employed the following detection criteria:
the non-starch polysaccharide is determined by subtracting the content of reducing sugar from the content of total sugar, and the total sugar and the reducing sugar are determined by adopting a nitro salicylic acid colorimetric method.
Accurately sucking 0mL, 0.2mL, 0.4mL, 0.8mL, 1.0mL and 1.2mL glucose standard solution into 6 10mL test tubes with plug scales respectively by a pipette, adding water to make the volume of the solution up to 2.0mL, adding 4.0mL 3, 5-dinitrosalicylic acid reagent, heating in a boiling water bath for 5min, taking out, immediately placing in cold water, cooling to room temperature, fixing the volume, and shaking up. The concentrations of the obtained serial dextran standard solutions were 0mg/mL, 0.02mg/mL, 0.04mg/mL, 0.08mg/mL, 0.10mg/mL, and 0.12mg/mL, respectively, and 540nm absorbance values were measured with a spectrophotometer. A standard curve was drawn with the glucose concentration (mg/mL) as ordinate (y) and the absorbance value as abscissa (x). According to the content of sugar in different samples, 5-20 mL (V) of filtrate is absorbed by a pipette2) In a volumetric flask, the volume is adjusted to 100mL (V) by water1) Draw 1.00mL (V) from the measuring flask1) The samples were placed in 10mL (Vs) volumetric flasks or cuvettes with stoppered scales, and water was added to 2.0mL each, following the procedure described above for the standard curve. The absorbance value measured was recorded, and the concentration of reducing sugar in the measurement solution was determined from the standard curve.
According to the sugar content of the sample, a pipette is used for sucking 5mL to 10mL (V) of sample liquid2) Adding 6mol/L hydrochloric acid solution 1mL into a volumetric flask, heating at 80 + -2 deg.C for 10min in a constant temperature water bath, taking out, cooling to room temperature in a cold water tank, adding 3 drops of methyl red indicator, neutralizing with 6mol/L sodium hydroxide solution to light orange, and diluting to 100mL (V/L) with water to constant volume3) And (4) uniformly mixing. The procedure for the determination of reducing sugars was as follows.
Blank test in which the same measurement procedure was used except that no sample was added, and the operation was carried out in parallel.
Calculation of results
The content of soluble sugar in the sample except the dried fruit product is calculated according to the formula (1) by mass fraction, and the content of soluble sugar in the dried fruit product is calculated according to the formula (2) by mass fraction.
Figure RE-GDA0003175187300000131
X-soluble sugar content in mass percent (%) in the sample;
p-sample determination of reducing sugar concentration in milligrams per milliliter (mg/mL); v1-sample volume in milliliters (mL);
V2sample volume in milliliters (mL);
V3-aliquoting the volumetric volume of the sample solution in milliliters (mL);
V4-assay aspirate volume in milliliters (mL);
V5-determining the volume of the sample solution in milliliters (mL);
a-dilution multiple, 1 for juicy fruits and 2 for fruits with low water content;
m-sample mass in grams (g);
10-conversion factor of the measurement results converted into mass percent.
Figure RE-GDA0003175187300000141
m1-sample pre-oven mass in grams (g);
m2the dried mass of the sample in grams (g).
The calculation result is represented to the two last decimal places.
Protein content was measured according to the method for measuring protein in a food.
According to the national standard GB 5009.5-2016, 0.2 g-2 g of fully and uniformly mixed solid sample, 2 g-5 g of semi-solid sample or 10g-25g of liquid sample (about 30 mg-40 mg of nitrogen) are weighed and accurately weighed to 0.001g, and then 0.4g of copper sulfate, 6g of potassium sulfate and 20mL of sulfuric acid are added into a digestion furnace for digestion. And after the temperature of the digestion furnace reaches 420 ℃, continuously digesting for 1h, taking out the liquid in the digestion tube to be green and transparent, cooling, adding 50mL of water, and automatically adding liquid, distilling, titrating and recording titration data on an automatic Kjeldahl apparatus (adding sodium hydroxide solution, hydrochloric acid or sulfuric acid standard solution and boric acid solution containing a mixed indicator A or B before use).
The protein content in the sample was calculated according to formula (1):
Figure RE-GDA0003175187300000142
(1) in the formula:
x-the amount of protein in the sample in grams per hundred grams (g/100 g);
V1-volume of test solution consumed sulfuric acid or hydrochloric acid standard titration solution in milliliters (mL);
V2-reagent blank consumes volume of sulfuric acid or hydrochloric acid standard titration solution in milliliters (mL)
c-concentration of sulfuric acid or hydrochloric acid standard titration solution in moles per liter (mol/L);
0.0140-1.0 mL sulfuric acid [ c (12H)2SO4)＝1.000mol/L]Or hydrochloric acid [ c (HCl) ═ 1.000mol/L]Mass of nitrogen equivalent to standard titration solution in grams (g);
m-mass of sample in grams (g);
V3volume of digestive juice aspirated in milliliters (mL);
f-the coefficient of converting nitrogen into protein, and the coefficient of converting nitrogen in various foods is shown in appendix A;
100-conversion factor. When the protein content is more than or equal to 1g/100g, three effective figures are reserved; at protein content <1g/100g, two significant digits are retained.
Note that when only the nitrogen content is detected, it is not necessary to multiply the protein conversion factor F.
Essential amino acids were determined according to the method for determining amino acids in food.
According to the national standard GB5009.235-2016, 1.00g (or 1.0mL) of sample is weighed into a 50mL volumetric flask, diluted to the mark by adding water and mixed uniformly. 0mL, 0.05mL, 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL (equivalent to NH) of the standard solution for precise ammonia nitrogen extraction3-N0μg、5.0μg、10.0μg、20.0μg40.0. mu.g, 60.0. mu.g, 80.0. mu.g, 100.0. mu.g) in 10mL cuvettes, respectively. 4mL of sodium acetate-acetic acid buffer solution (pH4.8) and 4mL of color developing agent were added to each cuvette, diluted to the scale with water, and mixed well. Heating in 100 deg.C water bath for 15min, taking out, cooling to room temperature, transferring into 1cm cuvette, measuring absorbance at wavelength of 400nm with zero tube as reference, and drawing standard curve or calculating linear regression equation. 2mL of the sample diluted solution was precisely aspirated into a 10mL colorimetric tube. 4mL of sodium acetate-acetic acid buffer solution (pH4.8) and 4mL of color developing agent are added, diluted to the scale with water, and mixed well. Heating in 100 deg.C water bath for 15min, taking out, cooling to room temperature, transferring into 1cm cuvette, and measuring absorbance at wavelength of 400nm with reference to zero tube.
And comparing the absorbance of the sample with the standard curve for quantification or substituting the absorbance into a linear regression equation to calculate the content of the sample. The content of amino acid nitrogen in the sample was calculated as follows:
Figure RE-GDA0003175187300000161
Figure RE-GDA0003175187300000162
X1-the content of amino acid nitrogen in the sample in grams per hundred grams (g/100 g);
X2-the content of amino acid nitrogen in the sample in grams per hundred milliliters (g/100 mL);
m-mass of nitrogen in the sample measurement solution in units of micrograms (ug);
m1weighing the mass of the sample, wherein the unit is gram (g);
v — volume of aspirated sample in milliliters (mL);
V1volume of sample solution for determination in milliliters (mL);
V2volume fixed in milliliters (mL) for sample pretreatment;
100. 1000-unit conversion factor.
Detecting the content of viable bacteria of beneficial bacteria, weighing 25g of sample by aseptic operation, placing the sample into an aseptic homogenizing cup filled with 225mL of physiological saline, and homogenizing for 1-2 min at 8000-10000 r/min to prepare 1:10 sample homogeneous solution; sucking 1:10 sample homogenizing solution 1mL by using a 1mL sterile sucker or a micropipette, slowly injecting the sample homogenizing solution into a sterile test tube filled with 9mL physiological saline along the tube wall, and fully and uniformly mixing by vortex oscillation to prepare the sample homogenizing solution 1: 100. Taking another 1mL sterile pipette or micropipette suction head, according to the above operation sequence, making 10 times of progressively increased sample homogenizing liquid, and changing 1mL sterile pipette or suction head for every time of progressively increased dilution. According to the estimation of the total viable count of the sample to be detected, 3 continuous suitable dilutions are selected, 0.1mL of sample homogenate is absorbed by each dilution and is respectively placed on 2 agar plates, and an L-shaped rod is used for surface coating. And (5) turning the plate, putting the plate into an incubator at 30 +/-1 ℃ for culturing for 20-48 h, and then calculating the number of all colonies on the plate. The dilution of the sample to plate coating was completed in 30 min. And (4) calculating a result:
Figure RE-GDA0003175187300000171
the viable bacteria content of the product is the total viable bacteria number obtained by detection.
Reports are made from colony counts, weighing samples are reported in CFU/g, volume samples are reported in CFU/mL.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and the present invention shall be covered thereby. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (9)

1. The wet soybean meal fermented by pediococcus acidilactici with high digestibility is characterized in that: adding liquid proliferated pediococcus acidilactici and complex enzyme preparation into pulverized soybean meal raw material, and fermenting.
2. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 1, wherein: the method specifically comprises the following steps: crushing the soybean meal raw material, adding water, uniformly mixing, sterilizing, simultaneously adding the liquid proliferated pediococcus acidilactici and the complex enzyme preparation, carrying out constant-temperature slow enzymolysis fermentation, carrying out solid-liquid separation after fermentation, drying and crushing to obtain the pediococcus acidilactici high-digestibility fermented wet soybean meal.
3. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 2, wherein: the mass ratio of the soybean meal raw material to water is 5-7: 3-5, wherein the conditions of constant-temperature slow enzymolysis and fermentation are 37 ℃ and 72 hours.
4. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 2, wherein: the complex enzyme preparation, namely the acidic protease, the neutral protease and the alkaline protease, comprises the following components in parts by mass: 2: 4, mixing.
5. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 2, wherein: the inoculation amount of the pediococcus acidilactici is 5-15%.
6. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 2, wherein: the addition amount of the acidic protease, the neutral protease and the alkaline protease is 2500U.
7. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 2, wherein: the pediococcus acidilactici (pediococcus acidilactici) is derived from a sample collected in a manual pickled Chinese cabbage workshop, is preserved in China general microbiological culture Collection center (CGMCC) at 9 and 16 days 2020, and has a preservation unit address as follows: no. 3 Xilu No. 1 Beijing, Chaoyang, and the preservation number is CGMCC No. 20658.
8. The high digestibility pediococcus acidilactici fermented wet soybean meal according to claim 7, wherein: the pediococcus acidilactici proliferated in liquid state is prepared by inoculating pediococcus acidilactici preserved in a microorganism strain preservation center into an MRS liquid culture medium according to the inoculation amount of 1 ‰, culturing at 37 deg.C for 24 hr, and activating for 2-3 generations.
9. The method for preparing the wet soybean meal fermented by pediococcus acidilactici according to claim 1, which is characterized by comprising the following steps: crushing the soybean meal raw material, adding water, uniformly mixing, sterilizing, simultaneously adding the pediococcus acidilactici and the compound protease which are proliferated in a liquid state, fermenting at constant temperature at a low speed, performing solid-liquid separation after fermentation, drying and crushing to obtain the pediococcus acidilactici high-digestibility fermented wet soybean meal.
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